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Electroporation

From Wikipedia, in a visual modern way
Cuvettes for in-vitro electroporation. These are plastic with aluminium electrodes and a blue lid. They hold a maximum of 400 μl.
Cuvettes for in-vitro electroporation. These are plastic with aluminium electrodes and a blue lid. They hold a maximum of 400 μl.

Electroporation, or electropermeabilization, is a microbiology technique in which an electrical field is applied to cells in order to increase the permeability of the cell membrane, allowing chemicals, drugs, electrode arrays or DNA to be introduced into the cell (also called electrotransfer).[1][2][3] In microbiology, the process of electroporation is often used to transform bacteria, yeast, or plant protoplasts by introducing new coding DNA. If bacteria and plasmids are mixed together, the plasmids can be transferred into the bacteria after electroporation, though depending on what is being transferred, cell-penetrating peptides or CellSqueeze could also be used. Electroporation works by passing thousands of volts (~8 kV/cm) across suspended cells in an electroporation cuvette.[1] Afterwards, the cells have to be handled carefully until they have had a chance to divide, producing new cells that contain reproduced plasmids. This process is approximately ten times more effective in increasing cell membrane's permeability than chemical transformation.[4]

Electroporation is also highly efficient for the introduction of foreign genes into tissue culture cells, especially mammalian cells. For example, it is used in the process of producing knockout mice, as well as in tumor treatment, gene therapy, and cell-based therapy. The process of introducing foreign DNA into eukaryotic cells is known as transfection. Electroporation is highly effective for transfecting cells in suspension using electroporation cuvettes. Electroporation has proven efficient for use on tissues in vivo, for in utero applications as well as in ovo transfection. Adherent cells can also be transfected using electroporation, providing researchers with an alternative to trypsinizing their cells prior to transfection. One downside to electroporation, however, is that after the process the gene expression of over 7,000 genes can be affected.[5] This can cause problems in studies where gene expression has to be controlled to ensure accurate and precise results.

Although bulk electroporation has many benefits over physical delivery methods such as microinjections and gene guns, it still has limitations, including low cell viability. Miniaturization of electroporation has been studied, leading to microelectroporation and nanotransfection of tissue utilizing electroporation-based techniques via nanochannels to minimally invasively deliver cargo to the cells.[6]

Electroporation has also been used as a mechanism to trigger cell fusion. Artificially induced cell fusion can be used to investigate and treat different diseases, like diabetes,[7][8][9] regenerate axons of the central nerve system,[10] and produce cells with desired properties, such as in cell vaccines for cancer immunotherapy.[11] However, the first and most known application of cell fusion is production of monoclonal antibodies in hybridoma technology, where hybrid cell lines (hybridomas) are formed by fusing specific antibody-producing B lymphocytes with a myeloma (B lymphocyte cancer) cell line.[12]

Discover more about Electroporation related topics

Microbiology

Microbiology

Microbiology is the scientific study of microorganisms, those being unicellular, multicellular, or acellular. Microbiology encompasses numerous sub-disciplines including virology, bacteriology, protistology, mycology, immunology, and parasitology.

Cell membrane

Cell membrane

The cell membrane is a biological membrane that separates and protects the interior of all cells from the outside environment. The cell membrane consists of a lipid bilayer, made up of two layers of phospholipids with cholesterols interspersed between them, maintaining appropriate membrane fluidity at various temperatures. The membrane also contains membrane proteins, including integral proteins that span the membrane and serve as membrane transporters, and peripheral proteins that loosely attach to the outer (peripheral) side of the cell membrane, acting as enzymes to facilitate interaction with the cell's environment. Glycolipids embedded in the outer lipid layer serve a similar purpose. The cell membrane controls the movement of substances in and out of cells and organelles, being selectively permeable to ions and organic molecules. In addition, cell membranes are involved in a variety of cellular processes such as cell adhesion, ion conductivity, and cell signalling and serve as the attachment surface for several extracellular structures, including the cell wall and the carbohydrate layer called the glycocalyx, as well as the intracellular network of protein fibers called the cytoskeleton. In the field of synthetic biology, cell membranes can be artificially reassembled.

DNA

DNA

Deoxyribonucleic acid is a polymer composed of two polynucleotide chains that coil around each other to form a double helix. The polymer carries genetic instructions for the development, functioning, growth and reproduction of all known organisms and many viruses. DNA and ribonucleic acid (RNA) are nucleic acids. Alongside proteins, lipids and complex carbohydrates (polysaccharides), nucleic acids are one of the four major types of macromolecules that are essential for all known forms of life.

Plant

Plant

Plants are predominantly photosynthetic eukaryotes, forming the kingdom Plantae. Most of them are multicellular. Historically, the plant kingdom encompassed all living things that were not animals, and included algae and fungi. All current definitions exclude the fungi and some of the algae. By one definition, plants form the clade Viridiplantae which consists of the green algae and the Embryophyta or land plants. The latter include the hornworts, liverworts, mosses, lycophytes, ferns, conifers and other gymnosperms, and the flowering plants. A definition based on genomes includes the Viridiplantae, along with the red algae and the glaucophytes, in the clade Archaeplastida.

Protoplast

Protoplast

Protoplast, is a biological term coined by Hanstein in 1880 to refer to the entire cell, excluding the cell wall. Protoplasts can be generated by stripping the cell wall from plant, bacterial, or fungal cells by mechanical, chemical or enzymatic means.

Plasmid

Plasmid

A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that benefit the survival of the organism and confer selective advantage such as antibiotic resistance. While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain only additional genes that may be useful in certain situations or conditions. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. In the laboratory, plasmids may be introduced into a cell via transformation. Synthetic plasmids are available for procurement over the internet.

Gene

Gene

In biology, the word gene can have several different meanings. The Mendelian gene is a basic unit of heredity and the molecular gene is a sequence of nucleotides in DNA that is transcribed to produce a functional RNA. There are two types of molecular genes: protein-coding genes and noncoding genes.

Mammal

Mammal

A mammal is a vertebrate animal of the class Mammalia. Mammals are characterized by the presence of milk-producing mammary glands for feeding their young, a neocortex region of the brain, fur or hair, and three middle ear bones. These characteristics distinguish them from reptiles and birds, which they diverged from in the Carboniferous Period over 300 million years ago. Around 6,400 extant species of mammals have been described and divided into 29 orders.

Microinjection

Microinjection

Microinjection is the use of a glass micropipette to inject a liquid substance at a microscopic or borderline macroscopic level. The target is often a living cell but may also include intercellular space. Microinjection is a simple mechanical process usually involving an inverted microscope with a magnification power of around 200x.

Gene gun

Gene gun

In genetic engineering, a gene gun or biolistic particle delivery system is a device used to deliver exogenous DNA (transgenes), RNA, or protein to cells. By coating particles of a heavy metal with a gene of interest and firing these micro-projectiles into cells using mechanical force, an integration of desired genetic information can be introduced into desired cells. The technique involved with such micro-projectile delivery of DNA is often referred to as biolistics, short for "biological ballistics".

Cell fusion

Cell fusion

Cell fusion is an important cellular process in which several uninucleate cells combine to form a multinucleate cell, known as a syncytium. Cell fusion occurs during differentiation of myoblasts, osteoclasts and trophoblasts, during embryogenesis, and morphogenesis. Cell fusion is a necessary event in the maturation of cells so that they maintain their specific functions throughout growth.

Hybridoma technology

Hybridoma technology

Hybridoma technology is a method for producing large numbers of identical antibodies. This process starts by injecting a mouse with an antigen that provokes an immune response. A type of white blood cell, the B cell, produces antibodies that bind to the injected antigen. These antibody producing B-cells are then harvested from the mouse and, in turn, fused with immortal B cell cancer cells, a myeloma, to produce a hybrid cell line called a hybridoma, which has both the antibody-producing ability of the B-cell and the longevity and reproductivity of the myeloma. The hybridomas can be grown in culture, each culture starting with one viable hybridoma cell, producing cultures each of which consists of genetically identical hybridomas which produce one antibody per culture (monoclonal) rather than mixtures of different antibodies (polyclonal). The myeloma cell line that is used in this process is selected for its ability to grow in tissue culture and for an absence of antibody synthesis. In contrast to polyclonal antibodies, which are mixtures of many different antibody molecules, the monoclonal antibodies produced by each hybridoma line are all chemically identical.

Laboratory practice

Electroporation is performed with electroporators, purpose-built appliances that create an electrostatic field in a cell solution. The cell suspension is pipetted into a glass or plastic cuvette which has two aluminium electrodes on its sides. For bacterial electroporation, typically a suspension of around 50 microliters is used. Prior to electroporation, this suspension of bacteria is mixed with the plasmid to be transformed. The mixture is pipetted into the cuvette, the voltage and capacitance are set, and the cuvette is inserted into the electroporator. The process requires direct contact between the electrodes and the suspension. Immediately after electroporation, one milliliter of liquid medium is added to the bacteria (in the cuvette or in an Eppendorf tube), and the tube is incubated at the bacteria's optimal temperature for an hour or more to allow recovery of the cells and expression of the plasmid, followed by bacterial culture on agar plates.

The success of the electroporation depends greatly on the purity of the plasmid solution, especially on its salt content. Solutions with high salt concentrations might cause an electrical discharge (known as arcing), which often reduces the viability of the bacteria. For a further detailed investigation of the process, more attention should be paid to the output impedance of the porator device and the input impedance of the cells suspension (e.g. salt content).

Since the cell membrane is not able to pass current (except in ion channels), it acts as an electrical capacitor. Subjecting membranes to a high-voltage electric field results in their temporary breakdown, resulting in pores that are large enough to allow macromolecules (such as DNA) to enter or leave the cell.[13]

Additionally, electroporation can be used to increase permeability of cells during in Utero injections and surgeries. Particularly, the electroporation allows for a more efficient transfection of DNA, RNA, shRNA, and all nucleic acids into the cells of mice and rats. The success of in vivo electroporation depends greatly on voltage, repetition, pulses, and duration. Developing central nervous systems are most effective for in vivo electroporation due to the visibility of ventricles for injections of nucleic acids, as well as the increased permeability of dividing cells. Electroporation of injected in utero embryos is performed through the uterus wall, often with forceps-type electrodes to limit damage to the embryo.[14]

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Suspension (chemistry)

Suspension (chemistry)

In chemistry, a suspension is a heterogeneous mixture of a fluid that contains solid particles sufficiently large for sedimentation. The particles may be visible to the naked eye, usually must be larger than one micrometer, and will eventually settle, although the mixture is only classified as a suspension when and while the particles have not settled out.

Pipette

Pipette

A pipette is a laboratory tool commonly used in chemistry, biology and medicine to transport a measured volume of liquid, often as a media dispenser. Pipettes come in several designs for various purposes with differing levels of accuracy and precision, from single piece glass pipettes to more complex adjustable or electronic pipettes. Many pipette types work by creating a partial vacuum above the liquid-holding chamber and selectively releasing this vacuum to draw up and dispense liquid. Measurement accuracy varies greatly depending on the instrument.

Electrode

Electrode

An electrode is an electrical conductor used to make contact with a nonmetallic part of a circuit. Electrodes are essential parts of batteries that can consist of a variety of materials depending on the type of battery.

Plasmid

Plasmid

A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that benefit the survival of the organism and confer selective advantage such as antibiotic resistance. While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain only additional genes that may be useful in certain situations or conditions. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. In the laboratory, plasmids may be introduced into a cell via transformation. Synthetic plasmids are available for procurement over the internet.

Agar

Agar

Agar, or agar-agar, is a jelly-like substance consisting of polysaccharides obtained from the cell walls of some species of red algae, primarily from "ogonori" (Gracilaria) and "tengusa" (Gelidiaceae). As found in nature, agar is a mixture of two components, the linear polysaccharide agarose and a heterogeneous mixture of smaller molecules called agaropectin. It forms the supporting structure in the cell walls of certain species of algae and is released on boiling. These algae are known as agarophytes, belonging to the Rhodophyta phylum. The processing of food-grade agar removes the agaropectin, and the commercial product is essentially pure agarose.

Electric arc

Electric arc

An electric arc is an electrical breakdown of a gas that produces a prolonged electrical discharge. The current through a normally nonconductive medium such as air produces a plasma; the plasma may produce visible light. An arc discharge is characterized by a lower voltage than a glow discharge and relies on thermionic emission of electrons from the electrodes supporting the arc. An archaic term is voltaic arc, as used in the phrase "voltaic arc lamp".

Output impedance

Output impedance

The output impedance of an electrical network is the measure of the opposition to current flow (impedance), both static (resistance) and dynamic (reactance), into the load network being connected that is internal to the electrical source. The output impedance is a measure of the source's propensity to drop in voltage when the load draws current, the source network being the portion of the network that transmits and the load network being the portion of the network that consumes.

Input impedance

Input impedance

The input impedance of an electrical network is the measure of the opposition to current (impedance), both static (resistance) and dynamic (reactance), into the load that is external to the electrical source network. The input admittance is a measure of the load network's propensity to draw current. The source network is the portion of the network that transmits power, and the load network is the portion of the network that consumes power.

Salt

Salt

Salt is a mineral composed primarily of sodium chloride (NaCl), a chemical compound belonging to the larger class of salts; salt in the form of a natural crystalline mineral is known as rock salt or halite. Salt is present in vast quantities in seawater. The open ocean has about 35 g (1.2 oz) of solids per liter of sea water, a salinity of 3.5%.

In vitro and animal studies

In vivo gene electrotransfer was first described in 1991[15] and today there are many preclinical studies of gene electrotransfer. The method is used to deliver large variety of therapeutic genes for potential treatment of several diseases, such as: disorders in immune system, tumors, metabolic disorders, monogenetic diseases, cardiovascular diseases, analgesia….[16][17][18]

With regards to irreversible electroporation, the first successful treatment of malignant cutaneous tumors implanted in mice was completed in 2007 by a group of scientists who achieved complete tumor ablation in 12 out of 13 mice. They accomplished this by sending 80 pulses of 100 microseconds at 0.3 Hz with an electrical field magnitude of 2500 V/cm to treat the cutaneous tumors.[19] Currently, a number of companies, including AngioDynamics, Inc. and VoltMed, Inc., are continuing to develop and deploy irreversible electroporation-based technologies within clinical environments.

The first group to look at electroporation for medical applications was led by Lluis M Mir at the Institute Gustave Roussy. In this case, they looked at the use of reversible electroporation in conjunction with impermeable macromolecules. The first research looking at how nanosecond pulses might be used on human cells was conducted by researchers at Eastern Virginia Medical School and Old Dominion University, and published in 2003.[20]

Medical applications

Main article: Irreversible electroporation

The first medical application of electroporation was used for introducing poorly permeant anticancer drugs into tumor nodules.[21] Soon also gene electrotransfer became of special interest because of its low cost, easiness of realization and safety. Namely, viral vectors can have serious limitations in terms of immunogenicity and pathogenicity when used for DNA transfer.[22]

A higher voltage of electroporation was found in pigs to irreversibly destroy target cells within a narrow range while leaving neighboring cells unaffected, and thus represents a promising new treatment for cancer, heart disease and other disease states that require removal of tissue.[23] Irreversible electroporation (IRE) has since proven effective in treating human cancer, with surgeons at Johns Hopkins and other institutions now using the technology to treat pancreatic cancer previously thought to be unresectable.[24]

Also first phase I clinical trial of gene electrotransfer in patients with metastatic melanoma was reported.[25][26] Electroporation mediated delivery of a plasmid coding gene for interleukin-12 (pIL-12) was performed and safety, tolerability and therapeutic effect were monitored. Study concluded, that gene electrotransfer with pIL-12 is safe and well tolerated. In addition partial or complete response was observed also in distant non treated metastases, suggesting the systemic treatment effect. Based on these results they are already planning to move to Phase II clinical study. There are currently several ongoing clinical studies of gene electrotransfer[27] where safety, tolerability and effectiveness of immunization with DNA vaccine, which is administered by the electric pulses is monitored.

Although the method is not systemic, but strictly local one, it is still the most efficient non-viral strategy for gene delivery.

N-TIRE

A recent technique called non-thermal irreversible electroporation (N-TIRE) has proven successful in treating many different types of tumors and other unwanted tissue. This procedure is done using small electrodes (about 1mm in diameter), placed either inside or surrounding the target tissue to apply short, repetitive bursts of electricity at a predetermined voltage and frequency. These bursts of electricity increase the resting transmembrane potential (TMP), so that nanopores form in the plasma membrane. When the electricity applied to the tissue is above the electric field threshold of the target tissue, the cells become permanently permeable from the formation of nanopores. As a result, the cells are unable to repair the damage and die due to a loss of homeostasis.[28] N-TIRE is unique to other tumor ablation techniques in that it does not create thermal damage to the tissue around it.

Reversible electroporation

Contrastingly, reversible electroporation occurs when the electricity applied with the electrodes is below the electric field threshold of the target tissue. Because the electricity applied is below the cells' threshold, it allows the cells to repair their phospholipid bilayer and continue on with their normal cell functions. Reversible electroporation is typically done with treatments that involve getting a drug or gene (or other molecule that is not normally permeable to the cell membrane) into the cell. Not all tissue has the same electric field threshold; therefore careful calculations need to be made prior to a treatment to ensure safety and efficacy.[29]

One major advantage of using N-TIRE is that, when done correctly according to careful calculations, it only affects the target tissue. Proteins, the extracellular matrix, and critical structures such as blood vessels and nerves are all unaffected and left healthy by this treatment. This allows for a quicker recovery, and facilitates a more rapid replacement of dead tumor cells with healthy cells.[30]

Before doing the procedure, scientists must carefully calculate exactly what needs to be done and treat each patient on an individual case-by-case basis. To do this, imaging technology such as CT scans and MRI's are commonly used to create a 3D image of the tumor. From this information, they can approximate the volume of the tumor and decide on the best course of action including the insertion site of electrodes, the angle they are inserted in, the voltage needed, and more, using software technology. Often, a CT machine will be used to help with the placement of electrodes during the procedure, particularly when the electrodes are being used to treat tumors in the brain.[31]

The entire procedure is very quick, typically taking about five minutes. The success rate of these procedures is high[32] and is very promising for future treatment in humans. One disadvantage to using N-TIRE is that the electricity delivered from the electrodes can stimulate muscle cells to contract, which could have lethal consequences depending on the situation. Therefore, a paralytic agent must be used when performing the procedure. The paralytic agents that have been used in such research are successful; however, there is always some risk, albeit slight, when using anesthetics.

H-FIRE

A more recent technique has been developed called high-frequency irreversible electroporation (H-FIRE). This technique uses electrodes to apply bipolar bursts of electricity at a high frequency, as opposed to unipolar bursts of electricity at a low frequency. This type of procedure has the same tumor ablation success as N-TIRE. However, it has one distinct advantage, H-FIRE does not cause muscle contraction in the patient and therefore there is no need for a paralytic agent.[33] Furthermore, H-FIRE has been demonstrated to produce more predictable ablations due to the lesser difference in the electrical properties of tissues at higher frequencies.[34]

Drug and gene delivery

Electroporation can also be used to help deliver drugs or genes into the cell by applying short and intense electric pulses that transiently permeabilize cell membrane, thus allowing transport of molecules otherwise not transported through a cellular membrane. This procedure is referred to as electrochemotherapy when the molecules to be transported are chemotherapeutic agents or gene electrotransfer when the molecule to be transported is DNA. Scientists from Karolinska Institutet and the University of Oxford use electroporation of exosomes to deliver siRNAs, antisense oligonucleotides, chemotherapeutic agents and proteins specifically to neurons after inject them systemically (in blood). Because these exosomes are able to cross the blood brain barrier, this protocol could solve the problem of poor delivery of medications to the central nervous system, and potentially treat Alzheimer's disease, Parkinson's disease, and brain cancer, among other conditions.[35]

Bacterial transformation is generally the easiest way to make large amounts of a particular protein needed for biotechnology purposes or in medicine. Since gene electrotransfer is very simple, rapid and highly effective technique it first became very convenient replacement for other transformation procedures.[36]

Recent research has shown that shock waves could be used for pre-treating the cell membrane prior to electroporation.[37][38] This synergistic strategy has shown to reduce external voltage requirement and create larger pores. Also application of shock waves allow scope to target desired membrane site. This procedure allows to control the size of the pore.

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Irreversible electroporation

Irreversible electroporation

Irreversible electroporation is a soft tissue ablation technique using short but strong electrical fields to create permanent and hence lethal nanopores in the cell membrane, to disrupt cellular homeostasis. The resulting cell death results from induced apoptosis or necrosis induced by either membrane disruption or secondary breakdown of the membrane due to transmembrane transfer of electrolytes and adenosine triphosphate. The main use of IRE lies in tumor ablation in regions where precision and conservation of the extracellular matrix, blood flow and nerves are of importance. The first generation of IRE for clinical use, in the form of the NanoKnife System, became commercially available for research purposes in 2009, solely for the surgical ablation of soft tissue tumors. Cancerous tissue ablation via IRE appears to show significant cancer specific immunological responses which are currently being evaluated alone and in combination with cancer immunotherapy.

Voltage

Voltage

Voltage, also known as electric pressure, electric tension, or (electric) potential difference, is the difference in electric potential between two points. In a static electric field, it corresponds to the work needed per unit of charge to move a test charge between the two points. In the International System of Units, the derived unit for voltage is named volt.

Johns Hopkins Hospital

Johns Hopkins Hospital

The Johns Hopkins Hospital (JHH) is the teaching hospital and biomedical research facility of the Johns Hopkins School of Medicine, located in Baltimore, Maryland, U.S. It was founded in 1889 using money from a bequest of over $7 million by city merchant, banker/financier, civic leader and philanthropist Johns Hopkins (1795–1873). Johns Hopkins Hospital and its School of Medicine are considered to be the founding institutions of modern American medicine and the birthplace of numerous famous medical traditions including rounds, residents and house staff. Many medical specialties were formed at the hospital including neurosurgery, by Harvey Cushing and Walter Dandy; cardiac surgery by Alfred Blalock; and child psychiatry, by Leo Kanner. Attached to the hospital is the Johns Hopkins Children’s Center which serves infants, children, teens, and young adults aged 0–21.

Pancreatic cancer

Pancreatic cancer

Pancreatic cancer arises when cells in the pancreas, a glandular organ behind the stomach, begin to multiply out of control and form a mass. These cancerous cells have the ability to invade other parts of the body. A number of types of pancreatic cancer are known.

Interleukin 12

Interleukin 12

Interleukin 12 (IL-12) is an interleukin that is naturally produced by dendritic cells, macrophages, neutrophils, and human B-lymphoblastoid cells (NC-37) in response to antigenic stimulation. IL-12 belongs to the family of interleukin-12. IL-12 family is unique in comprising the only heterodimeric cytokines, which includes IL-12, IL-23, IL-27 and IL-35. Despite sharing many structural features and molecular partners, they mediate surprisingly diverse functional effects.

Electrochemotherapy

Electrochemotherapy

Electrochemotherapy is a type of chemotherapy that allows delivery of non-permeant drugs to the cell interior. It is based on the local application of short and intense electric pulses that transiently permeabilize the cell membrane, thus allowing transport of molecules otherwise not permitted by the membrane. Applications for treatment of cutaneous and subcutaneous tumors have reached clinical use by utilizing drugs such as bleomycin or cisplatin). Electrochemotherapy with bleomycin was used to treat a patient for the first time in 1991 at the Institute Gustave Roussy in France, while electrochemotherapy with cisplatin was used to treat for the first time in 1995 at the Institute of Oncology, Ljubljana, Slovenia. Since then, more than 4000 patients were treated with electrochemotherapy all over the world. Recently, new electrochemotherapy modalities have been developed for treatment of internal tumors using surgical procedures, endoscopic routes, or percutaneous approaches to gain access to the treatment area.

University of Oxford

University of Oxford

The University of Oxford is a collegiate research university in Oxford, England. There is evidence of teaching as early as 1096, making it the oldest university in the English-speaking world and the world's second-oldest university in continuous operation. It grew rapidly from 1167 when Henry II banned English students from attending the University of Paris. After disputes between students and Oxford townsfolk in 1209, some academics fled north-east to Cambridge where they established what became the University of Cambridge. The two English ancient universities share many common features and are jointly referred to as Oxbridge.

Exosome complex

Exosome complex

The exosome complex is a multi-protein intracellular complex capable of degrading various types of RNA molecules. Exosome complexes are found in both eukaryotic cells and archaea, while in bacteria a simpler complex called the degradosome carries out similar functions.

Alzheimer's disease

Alzheimer's disease

Alzheimer's disease (AD) is a neurodegenerative disease that usually starts slowly and progressively worsens. It is the cause of 60–70% of cases of dementia. The most common early symptom is difficulty in remembering recent events. As the disease advances, symptoms can include problems with language, disorientation, mood swings, loss of motivation, self-neglect, and behavioral issues. As a person's condition declines, they often withdraw from family and society. Gradually, bodily functions are lost, ultimately leading to death. Although the speed of progression can vary, the typical life expectancy following diagnosis is three to nine years.

Parkinson's disease

Parkinson's disease

Parkinson's disease (PD), or simply Parkinson's, is a long-term degenerative disorder of the central nervous system that mainly affects the motor system. The symptoms usually emerge slowly, and as the disease worsens, non-motor symptoms become more common. The most obvious early symptoms are tremor, rigidity, slowness of movement, and difficulty with walking. Cognitive and behavioral problems may also occur with depression, anxiety, and apathy occurring in many people with PD. Parkinson's disease dementia becomes common in the advanced stages of the disease. Those with Parkinson's can also have problems with their sleep and sensory systems. The motor symptoms of the disease result from the death of nerve cells in the substantia nigra, a region of the midbrain, leading to a dopamine deficit. The cause of this cell death is poorly understood, but involves the build-up of misfolded proteins into Lewy bodies in the neurons. Collectively, the main motor symptoms are also known as parkinsonism or a parkinsonian syndrome.

Physical mechanism

Schematic cross-section showing the theoretical arrangement of lipids in a hydrophobic pore (top) and a hydrophilic pore (bottom).
Schematic cross-section showing the theoretical arrangement of lipids in a hydrophobic pore (top) and a hydrophilic pore (bottom).
Further information: Lipid bilayer mechanics

Electroporation allows cellular introduction of large highly charged molecules such as DNA which would never passively diffuse across the hydrophobic bilayer core.[1] This phenomenon indicates that the mechanism is the creation of nm-scale water-filled holes in the membrane.[39] Electropores were optically imaged in lipid bilayer models like droplet interface bilayers[40] and giant unilamellar vesicles,[41] while addition of cytoskeletal proteins such as actin networks to the giant unilamellar vesicles seem to prevent the formation of visible electropores.[42] Experimental evidences for actin networks in regulating the cell membrane permeability has also emerged.[43] Although electroporation and dielectric breakdown both result from application of an electric field, the mechanisms involved are fundamentally different. In dielectric breakdown the barrier material is ionized, creating a conductive pathway. The material alteration is thus chemical in nature. In contrast, during electroporation the lipid molecules are not chemically altered but simply shift position, opening up a pore which acts as the conductive pathway through the bilayer as it is filled with water.

Electroporation is a dynamic phenomenon that depends on the local transmembrane voltage at each point on the cell membrane. It is generally accepted that for a given pulse duration and shape, a specific transmembrane voltage threshold exists for the manifestation of the electroporation phenomenon (from 0.5 V to 1 V). This leads to the definition of an electric field magnitude threshold for electroporation (Eth). That is, only the cells within areas where E≧Eth are electroporated. If a second threshold (Eir) is reached or surpassed, electroporation will compromise the viability of the cells, i.e., irreversible electroporation (IRE).[44]

Electroporation is a multi-step process with several distinct phases.[45] First, a short electrical pulse must be applied. Typical parameters would be 300–400 mV for capacitor through the migration of ions from the surrounding solution. Once the critical field is achieved there is a rapid localized rearrangement in lipid morphology. The resulting structure is believed to be a "pre-pore" since it is not electrically conductive but leads rapidly to the creation of a conductive pore.[46] Evidence for the existence of such pre-pores comes mostly from the "flickering" of pores, which suggests a transition between conductive and insulating states.[47] It has been suggested that these pre-pores are small (~3 Å) hydrophobic defects. If this theory is correct, then the transition to a conductive state could be explained by a rearrangement at the pore edge, in which the lipid heads fold over to create a hydrophilic interface. Finally, these conductive pores can either heal, resealing the bilayer or expand, eventually rupturing it. The resultant fate depends on whether the critical defect size was exceeded[48] which in turn depends on the applied field, local mechanical stress and bilayer edge energy.

Gene electroporation

Application of electric pulses of sufficient strength to the cell causes an increase in the trans-membrane potential difference, which provokes the membrane destabilization. Cell membrane permeability is increased and otherwise nonpermeant molecules enter the cell.[49][50] Although the mechanisms of gene electrotransfer are not yet fully understood, it was shown that the introduction of DNA only occurs in the part of the membrane facing the cathode and that several steps are needed for successful transfection: electrophoretic migration of DNA towards the cell, DNA insertion into the membrane, translocation across the membrane, migration of DNA towards the nucleus, transfer of DNA across the nuclear envelope and finally gene expression.[51] There are a number of factors that can influence the efficiency of gene electrotransfer, such as: temperature, parameters of electric pulses, DNA concentration, electroporation buffer used, cell size and the ability of cells to express transfected genes.[52] In in vivo gene electrotransfer, DNA diffusion through extracellular matrix, properties of tissue and overall tissue conductivity are also crucial.[53]

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Lipid bilayer mechanics

Lipid bilayer mechanics

Lipid bilayer mechanics is the study of the physical material properties of lipid bilayers, classifying bilayer behavior with stress and strain rather than biochemical interactions. Local point deformations such as membrane protein interactions are typically modelled with the complex theory of biological liquid crystals but the mechanical properties of a homogeneous bilayer are often characterized in terms of only three mechanical elastic moduli: the area expansion modulus Ka, a bending modulus Kb and an edge energy . For fluid bilayers the shear modulus is by definition zero, as the free rearrangement of molecules within plane means that the structure will not support shear stresses. These mechanical properties affect several membrane-mediated biological processes. In particular, the values of Ka and Kb affect the ability of proteins and small molecules to insert into the bilayer. Bilayer mechanical properties have also been shown to alter the function of mechanically activated ion channels.

DNA

DNA

Deoxyribonucleic acid is a polymer composed of two polynucleotide chains that coil around each other to form a double helix. The polymer carries genetic instructions for the development, functioning, growth and reproduction of all known organisms and many viruses. DNA and ribonucleic acid (RNA) are nucleic acids. Alongside proteins, lipids and complex carbohydrates (polysaccharides), nucleic acids are one of the four major types of macromolecules that are essential for all known forms of life.

Lipid bilayer

Lipid bilayer

The lipid bilayer is a thin polar membrane made of two layers of lipid molecules. These membranes are flat sheets that form a continuous barrier around all cells. The cell membranes of almost all organisms and many viruses are made of a lipid bilayer, as are the nuclear membrane surrounding the cell nucleus, and membranes of the membrane-bound organelles in the cell. The lipid bilayer is the barrier that keeps ions, proteins and other molecules where they are needed and prevents them from diffusing into areas where they should not be. Lipid bilayers are ideally suited to this role, even though they are only a few nanometers in width, because they are impermeable to most water-soluble (hydrophilic) molecules. Bilayers are particularly impermeable to ions, which allows cells to regulate salt concentrations and pH by transporting ions across their membranes using proteins called ion pumps.

History

In the 1960s it was known that by applying an external electric field, a large membrane potential at the two pole of a cell can be created. In the 1970s it was discovered that when a membrane potential reached a critical level, the membrane would break down and that it could recover.[54] By the 1980s, this opening was being used to introduce various materials/molecules into the cells.[55]

Source: "Electroporation", Wikipedia, Wikimedia Foundation, (2023, February 17th), https://en.wikipedia.org/wiki/Electroporation.

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Further Reading

Electrophysiology

Lipid bilayer

Membrane potential

Transformation (genetics)

Gene delivery

Sonoporation

Nanoknife

Vectors in gene therapy

Genetic engineering techniques

Intracellular delivery

Hydrodynamic delivery
References
  1. ^ a b c Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH (1982). "Gene transfer into mouse lyoma cells by electroporation in high electric fields". The EMBO Journal. 1 (7): 841–5. doi:10.1002/j.1460-2075.1982.tb01257.x. PMC 553119. PMID 6329708.
  2. ^ Jimbo, Yasutoshi; Sasaki, Daisuke; Ohya, Takashi; Lee, Sunghoon; Lee, Wonryung; Arab Hassani, Faezeh; Yokota, Tomoyuki; Matsuura, Katsuhisa; Umezu, Shinjiro; Shimizu, Tatsuya; Someya, Takao (2021). "An organic transistor matrix for multipoint intracellular action potential recording". Proceedings of the National Academy of Sciences. 118 (39): e2022300118. Bibcode:2021PNAS..11822300J. doi:10.1073/pnas.2022300118. PMC 8488610. PMID 34544852. S2CID 237584521.
  3. ^ Chang, Donald C. (2006-09-15), "Electroporation and Electrofusion", in Meyers, Robert A. (ed.), Encyclopedia of Molecular Cell Biology and Molecular Medicine, Wiley-VCH Verlag GmbH & Co. KGaA, doi:10.1002/3527600906.mcb.200300026, ISBN 9783527600908
  4. ^ Sugar IP, Neumann E (May 1984). "Stochastic model for electric field-induced membrane pores. Electroporation". Biophysical Chemistry. 19 (3): 211–25. doi:10.1016/0301-4622(84)87003-9. PMID 6722274.
  5. ^ Anne Trafton (2 February 2016). "Cell squeezing enhances protein imaging". MIT News Office.
  6. ^ Gallego-Perez, Daniel; Ghatak, Subhadip; Pal, Durba (October 2017). "Topical tissue nano-transfection mediates non-viral stroma reprogramming and rescue". Nature Nanotechnology. 12 (10): 974–979. Bibcode:2017NatNa..12..974G. doi:10.1038/nnano.2017.134. ISSN 1748-3395. PMC 5814120. PMID 28785092.
  7. ^ McClenaghan NH (May 2007). "Physiological regulation of the pancreatic {beta}-cell: functional insights for understanding and therapy of diabetes". Experimental Physiology. 92 (3): 481–96. doi:10.1113/expphysiol.2006.034835. PMID 17272356. S2CID 22548866.
  8. ^ Yanai G, Hayashi T, Zhi Q, Yang KC, Shirouzu Y, Shimabukuro T, Hiura A, Inoue K, Sumi S (2013). "Electrofusion of mesenchymal stem cells and islet cells for diabetes therapy: a rat model". PLOS ONE. 8 (5): e64499. Bibcode:2013PLoSO...864499Y. doi:10.1371/journal.pone.0064499. PMC 3665804. PMID 23724055.
  9. ^ McCluskey JT, Hamid M, Guo-Parke H, McClenaghan NH, Gomis R, Flatt PR (June 2011). "Development and functional characterization of insulin-releasing human pancreatic beta cell lines produced by electrofusion". The Journal of Biological Chemistry. 286 (25): 21982–92. doi:10.1074/jbc.M111.226795. PMC 3121343. PMID 21515691.
  10. ^ Sretavan DW, Chang W, Hawkes E, Keller C, Kliot M (October 2005). "Microscale surgery on single axons". Neurosurgery. 57 (4): 635–46, discussion 635–46. doi:10.1227/01.NEU.0000175545.57795.ac. PMID 16239875. S2CID 196411777.
  11. ^ Takakura K, Kajihara M, Ito Z, Ohkusa T, Gong J, Koido S (March 2015). "Dendritic-tumor fusion cells in cancer immunotherapy". Discovery Medicine. 19 (104): 169–74. PMID 25828520.
  12. ^ Trontelj K, Rebersek M, Kanduser M, Serbec VC, Sprohar M, Miklavcic D (November 2008). "Optimization of bulk cell electrofusion in vitro for production of human-mouse heterohybridoma cells". Bioelectrochemistry. 74 (1): 124–9. doi:10.1016/j.bioelechem.2008.06.003. PMID 18667367.
  13. ^ Potter H (May 2003). "Transfection by electroporation". Current Protocols in Molecular Biology. Chapter 9: Unit 9.3. doi:10.1002/0471142727.mb0903s62. ISBN 978-0471142720. PMC 2975437. PMID 18265334.
  14. ^ Saito, Tetsuichiro (2010), "Embryonic In Vivo Electroporation in the Mouse", Guide to Techniques in Mouse Development, Part B: Mouse Molecular Genetics, 2nd Edition, Methods in Enzymology, vol. 477, Elsevier, pp. 37–50, doi:10.1016/s0076-6879(10)77003-8, ISBN 978-0-12-384880-2, PMID 20699135, retrieved 2022-09-19
  15. ^ Titomirov AV, Sukharev S, Kistanova E (January 1991). "In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA". Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression. 1088 (1): 131–4. doi:10.1016/0167-4781(91)90162-F. PMID 1703441.
  16. ^ Heller LC, Coppola D (October 2002). "Electrically mediated delivery of vector plasmid DNA elicits an antitumor effect". Gene Therapy. 9 (19): 1321–5. doi:10.1038/sj.gt.3301802. PMID 12224015.
  17. ^ Chuang IC, Jhao CM, Yang CH, Chang HC, Wang CW, Lu CY, Chang YJ, Lin SH, Huang PL, Yang LC (2004). "Intramuscular electroporation with the pro-opiomelanocortin gene in rat adjuvant arthritis". Arthritis Research & Therapy. 6 (1): R7–R14. doi:10.1186/ar1014. PMC 400409. PMID 14979933.
  18. ^ Vilquin JT, Kennel PF, Paturneau-Jouas M, Chapdelaine P, Boissel N, Delaère P, Tremblay JP, Scherman D, Fiszman MY, Schwartz K (July 2001). "Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies". Gene Therapy. 8 (14): 1097–107. doi:10.1038/sj.gt.3301484. PMID 11526457. S2CID 1081582.
  19. ^ Al-Sakere B, André F, Bernat C, Connault E, Opolon P, Davalos RV, Rubinsky B, Mir LM (November 2007). "Tumor ablation with irreversible electroporation". PLOS ONE. 2 (11): e1135. Bibcode:2007PLoSO...2.1135A. doi:10.1371/journal.pone.0001135. PMC 2065844. PMID 17989772.
  20. ^ Beebe SJ, Fox PM, Rec LJ, Willis EL, Schoenbach KH (August 2003). "Nanosecond, high-intensity pulsed electric fields induce apoptosis in human cells". FASEB Journal. 17 (11): 1493–5. doi:10.1096/fj.02-0859fje. PMID 12824299. S2CID 13189517.
  21. ^ Mir LM, Belehradek M, Domenge C, Orlowski S, Poddevin B, Belehradek J, Schwaab G, Luboinski B, Paoletti C (1991). "[Electrochemotherapy, a new antitumor treatment: first clinical trial]". Comptes Rendus de l'Académie des Sciences, Série III (in French). 313 (13): 613–8. PMID 1723647.
  22. ^ Marshall E (December 1999). "Gene therapy death prompts review of adenovirus vector". Science. 286 (5448): 2244–5. doi:10.1126/science.286.5448.2244. PMID 10636774. S2CID 46362535.
  23. ^ Sarah Yang (2007-02-12). "New medical technique punches holes in cells, could treat tumors". Retrieved 2007-12-13.
  24. ^ "A Potential Boon for Pancreatic Cancer Patients". Johns Hopkins Surgery: News from the Johns Hopkins Department of Surgery. 2014-06-23.
  25. ^ Daud AI, DeConti RC, Andrews S, Urbas P, Riker AI, Sondak VK, Munster PN, Sullivan DM, Ugen KE, Messina JL, Heller R (December 2008). "Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma". Journal of Clinical Oncology. 26 (36): 5896–903. doi:10.1200/JCO.2007.15.6794. PMC 2645111. PMID 19029422.
  26. ^ Cha E, Daud A (November 2012). "Plasmid IL-12 electroporation in melanoma". Human Vaccines & Immunotherapeutics. 8 (11): 1734–8. doi:10.4161/hv.22573. PMC 3601150. PMID 23151447.
  27. ^ "2 Studies found for: gene electrotransfer". ClinicalTrials.gov. National Library of Medicine. Retrieved 19 September 2022.
  28. ^ Garcia PA, Rossmeisl JH, Davalos RV (2011). "Electrical conductivity changes during irreversible electroporation treatment of brain cancer". Conference Proceedings. 2011: 739–42. doi:10.1109/IEMBS.2011.6090168. ISBN 978-1-4577-1589-1. PMID 22254416. S2CID 4953213.
  29. ^ Garcia PA, Neal RE, Rossmeisl JH, Davalos RV (2010). "Non-thermal irreversible electroporation for deep intracranial disorders". Conference Proceedings. 2010: 2743–6. doi:10.1109/IEMBS.2010.5626371. ISBN 978-1-4244-4123-5. PMID 21095962. S2CID 9589956.
  30. ^ Garcia PA, Rossmeisl JH, Neal RE, Ellis TL, Olson JD, Henao-Guerrero N, Robertson J, Davalos RV (July 2010). "Intracranial nonthermal irreversible electroporation: in vivo analysis". The Journal of Membrane Biology. 236 (1): 127–36. CiteSeerX 10.1.1.679.527. doi:10.1007/s00232-010-9284-z. PMID 20668843. S2CID 10958480.
  31. ^ Neal RE, Garcia PA, Rossmeisl JH, Davalos RV (2010). "A study using irreversible electroporation to treat large, irregular tumors in a canine patient". Conference Proceedings. 2010: 2747–50. doi:10.1109/IEMBS.2010.5626372. ISBN 978-1-4244-4123-5. PMID 21095963. S2CID 24348785.
  32. ^ Potter, H (2003). "Transfection by Electroporation". Current Protocols in Molecular Biology.
  33. ^ Arena CB, Sano MB, Rossmeisl JH, Caldwell JL, Garcia PA, Rylander MN, Davalos RV (November 2011). "High-frequency irreversible electroporation (H-FIRE) for non-thermal ablation without muscle contraction". BioMedical Engineering OnLine. 10: 102. doi:10.1186/1475-925X-10-102. PMC 3258292. PMID 22104372.
  34. ^ Bhonsle SP, Arena CB, Sweeney DC, Davalos RV (27 August 2015). "Mitigation of impedance changes due to electroporation therapy using bursts of high-frequency bipolar pulses". BioMedical Engineering OnLine. 13 (Suppl 3): S3. doi:10.1186/1475-925X-14-S3-S3. PMC 4565149. PMID 26355870.
  35. ^ El-Andaloussi S, Lee Y, Lakhal-Littleton S, Li J, Seow Y, Gardiner C, Alvarez-Erviti L, Sargent IL, Wood MJ (December 2012). "Exosome-mediated delivery of siRNA in vitro and in vivo". Nature Protocols. 7 (12): 2112–26. doi:10.1038/nprot.2012.131. PMID 23154783. S2CID 34413410.
  36. ^ Calvin NM, Hanawalt PC (June 1988). "High-efficiency transformation of bacterial cells by electroporation". Journal of Bacteriology. 170 (6): 2796–801. doi:10.1128/jb.170.6.2796-2801.1988. PMC 211205. PMID 3286620.
  37. ^ Hu, Q; Hossain, S; Joshi, R P (2018-06-25). "Analysis of a dual shock-wave and ultrashort electric pulsing strategy for electro-manipulation of membrane nanopores". Journal of Physics D: Applied Physics. 51 (28): 285403. Bibcode:2018JPhD...51B5403H. doi:10.1088/1361-6463/aaca7a. ISSN 0022-3727. S2CID 125134522.
  38. ^ Hossain, Shadeeb; Abdelgawad, Ahmed (2020-01-02). "Analysis of membrane permeability due to synergistic effect of controlled shock wave and electric field application". Electromagnetic Biology and Medicine. 39 (1): 20–29. doi:10.1080/15368378.2019.1706553. ISSN 1536-8378. PMID 31868023. S2CID 209446699.
  39. ^ Chang DC, Reese TS (July 1990). "Changes in membrane structure induced by electroporation was revealed by rapid-freezing electron microscopy". Biophysical Journal. 58 (1): 1–12. Bibcode:1990BpJ....58....1C. doi:10.1016/S0006-3495(90)82348-1. PMC 1280935. PMID 2383626.
  40. ^ Sengel, Jason T.; Wallace, Mark I. (10 May 2016). "Imaging the dynamics of individual electropores". Proceedings of the National Academy of Sciences. 113 (19): 5281–5286. Bibcode:2016PNAS..113.5281S. doi:10.1073/pnas.1517437113. PMC 4868429. PMID 27114528.
  41. ^ Sachdev, Shaurya; Muralidharan, Aswin; Choudhary, Dipendra K.; Perrier, Dayinta L.; Rems, Lea; Kreutzer, Michiel T.; Boukany, Pouyan E. (2019). "DNA translocation to giant unilamellar vesicles during electroporation is independent of DNA size". Soft Matter. 15 (45): 9187–9194. Bibcode:2019SMat...15.9187S. doi:10.1039/C9SM01274E. PMID 31595286.
  42. ^ Perrier, Dayinta L.; Vahid, Afshin; Kathavi, Vaishnavi; Stam, Lotte; Rems, Lea; Mulla, Yuval; Muralidharan, Aswin; Koenderink, Gijsje H.; Kreutzer, Michiel T.; Boukany, Pouyan E. (31 May 2019). "Response of an actin network in vesicles under electric pulses". Scientific Reports. 9 (1): 8151. Bibcode:2019NatSR...9.8151P. doi:10.1038/s41598-019-44613-5. PMC 6544639. PMID 31148577.
  43. ^ Muralidharan, Aswin; Rems, Lea; Kreutzer, Michiel T.; Boukany, Pouyan E. (August 2020). "Actin networks regulate the cell membrane permeability during electroporation". Biochimica et Biophysica Acta (BBA) - Biomembranes. 1863 (1): 183468. doi:10.1016/j.bbamem.2020.183468. PMID 32882211.
  44. ^ Ivorra, Antoni; Rubinsky, Boris. "Gels with predetermined conductivity used in electroporation of tissue USPTO Application #: 20080214986 — Class: 604 21 (USPTO)". Archived from the original on 2014-10-22. Retrieved 2008-11-21.
  45. ^ Ho SY, Mittal GS (1996). "Electroporation of cell membranes: a review". Critical Reviews in Biotechnology. 16 (4): 349–62. doi:10.3109/07388559609147426. PMID 8989868.
  46. ^ Becker SM, Kuznetsov AV (October 2007). "Local temperature rises influence in vivo electroporation pore development: a numerical stratum corneum lipid phase transition model". Journal of Biomechanical Engineering. 129 (5): 712–21. doi:10.1115/1.2768380. PMID 17887897.
  47. ^ Melikov KC, Frolov VA, Shcherbakov A, Samsonov AV, Chizmadzhev YA, Chernomordik LV (April 2001). "Voltage-induced nonconductive pre-pores and metastable single pores in unmodified planar lipid bilayer". Biophysical Journal. 80 (4): 1829–36. Bibcode:2001BpJ....80.1829M. doi:10.1016/S0006-3495(01)76153-X. PMC 1301372. PMID 11259296.
  48. ^ Joshi RP, Schoenbach KH (July 2000). "Electroporation dynamics in biological cells subjected to ultrafast electrical pulses: a numerical simulation study". Physical Review E. 62 (1 Pt B): 1025–33. Bibcode:2000PhRvE..62.1025J. doi:10.1103/PhysRevE.62.1025. PMID 11088559.
  49. ^ Kotnik, T; Miklavcic, D (August 2000). "Analytical description of transmembrane voltage induced by electric fields on spheroidal cells". Biophys J. 79 (2): 670–9. Bibcode:2000BpJ....79..670K. doi:10.1016/S0006-3495(00)76325-9. PMC 1300967. PMID 10920001.
  50. ^ Sweeney DC, Weaver JC, Davalos RV (January 2018). "Characterization of Cell Membrane Permeability In Vitro Part I: Transport Behavior Induced by Single-Pulse Electric Fields". Technology in Cancer Research & Treatment. 17: 1533033818792491. doi:10.1177/1533033818792491. PMC 6154305. PMID 30236040.
  51. ^ Satkauskas S, Bureau MF, Puc M, Mahfoudi A, Scherman D, Miklavcic D, Mir LM (February 2002). "Mechanisms of in vivo DNA electrotransfer: respective contributions of cell electropermeabilization and DNA electrophoresis". Molecular Therapy. 5 (2): 133–40. doi:10.1006/mthe.2002.0526. PMID 11829520.
  52. ^ Gehl J (April 2003). "Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research". Acta Physiologica Scandinavica. 177 (4): 437–47. doi:10.1046/j.1365-201X.2003.01093.x. PMID 12648161. S2CID 16742681.
  53. ^ Miklavcic D, Beravs K, Semrov D, Cemazar M, Demsar F, Sersa G (May 1998). "The importance of electric field distribution for effective in vivo electroporation of tissues". Biophysical Journal. 74 (5): 2152–8. Bibcode:1998BpJ....74.2152M. doi:10.1016/S0006-3495(98)77924-X. PMC 1299558. PMID 9591642.
  54. ^ Guide to electroporation and electrofusion. Donald C. Chang. San Diego: Academic Press. 1992. ISBN 978-0-12-168041-1. OCLC 817706277.{{cite book}}: CS1 maint: others (link)
  55. ^ Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH (1982). "Gene transfer into mouse lyoma cells by electroporation in high electric fields". The EMBO Journal. 1 (7): 841–5. doi:10.1002/j.1460-2075.1982.tb01257.x. PMC 553119. PMID 6329708.
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