Wikipedia

Cell culture

"Co-culture" redirects here. For the concept of cultures-within-cultures, seeSubculture.

Cell cultureortissue cultureis the process by whichcellsare grown under controlled conditions, generally outside of their natural environment. After cells of interest have beenisolated from living tissue,they can subsequently be maintained under carefully controlled conditions. They need to be kept at body temperature (37 °C) in an incubator.[1]These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or richmediumthat supplies the essential nutrients (amino acids,carbohydrates,vitamins,minerals),growth factors,hormones,and gases (CO2,O2), and regulates the physio-chemical environment (pH buffer,osmotic pressure,temperature). Most cells require a surface or an artificial substrate to form anadherent cultureas a monolayer (one single-cell thick), whereas others can be grown free floating in a medium as asuspension culture.[2]This is typically facilitated via use of a liquid, semi-solid, or solidgrowth medium,such asbrothoragar.Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific termplant tissue culturebeing used for plants. The lifespan of most cells is genetically determined, but some cell-culturing cells have been 'transformed' into immortal cells which will reproduce indefinitely if the optimal conditions are provided.

Cell culture in a smallPetri dish
Epithelial cellsin culture,stainedforkeratin(red) andDNA(green)

In practice, the term "cell culture" now refers to the culturing of cells derived from multicellulareukaryotes,especially animal cells, in contrast with other types of culture that also grow cells, such asplant tissue culture,fungalculture, andmicrobiological culture(ofmicrobes). The historical development and methods of cell culture are closely interrelated with those oftissue cultureandorgan culture.Viral cultureis also related, with cells as hosts for the viruses.

The laboratory technique of maintaining livecell lines(a population of cells descended from a single cell and containing the same genetic makeup) separated from their original tissue source became more robust in the middle 20th century.[3][4]

History

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The 19th-century English physiologistSydney Ringerdevelopedsalt solutionscontaining the chlorides of sodium, potassium, calcium and magnesium suitable for maintaining the beating of an isolatedanimal heartoutside the body.[5]In 1885Wilhelm Rouxremoved a section of themedullary plateof anembryonicchickenand maintained it in a warmsaline solutionfor several days, establishing the basic principle of tissue culture. In 1907 the zoologistRoss Granville Harrisondemonstrated the growth of frog embryonic cells that would give rise to nerve cells in a medium of clottedlymph.In 1913, E. Steinhardt, C. Israeli, and R. A. Lambert grewvacciniavirusin fragments of guinea pigcornealtissue.[6]In 1996, the first use of regenerative tissue was used to replace a small length of urethra, which led to the understanding that the technique of obtaining samples of tissue, growing it outside the body without a scaffold, and reapplying it, can be used for only small distances of less than 1 cm.[7][8][9]Ross Granville Harrison,working atJohns Hopkins Medical Schooland then atYale University,published results of his experiments from 1907 to 1910, establishing the methodology oftissue culture.[10]

Gottlieb Haberlandtfirst pointed out the possibilities of the culture of isolated tissues,plant tissue culture.[11]He suggested that the potentialities of individual cells via tissue culture as well as that the reciprocal influences of tissues on one another could be determined by this method. Since Haberlandt's original assertions, methods for tissue and cell culture have been realized, leading to significant discoveries in biology and medicine. He presented his original idea oftotipotentialityin 1902, stating that "Theoretically all plant cells are able to give rise to a complete plant."[12][13][14]The termtissue culturewas coined by American pathologistMontrose Thomas Burrows.[15]

Cell culture techniques were advanced significantly in the 1940s and 1950s to support research invirology.Growing viruses in cell cultures allowed preparation of purified viruses for the manufacture ofvaccines.The injectablepolio vaccinedeveloped byJonas Salkwas one of the first products mass-produced using cell culture techniques. This vaccine was made possible by the cell culture research ofJohn Franklin Enders,Thomas Huckle Weller,andFrederick Chapman Robbins,who were awarded aNobel Prizefor their discovery of a method of growing the virus in monkeykidneycell cultures. Cell culture has contributed to the development of vaccines for many diseases.[1]

Modern usage

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Cultured cells growing ingrowth medium

In modern usage, "tissue culture" generally refers to the growth of cells from a tissue from amulticellularorganismin vitro.These cells may be cells isolated from a donor organism (primary cells) or animmortalised cell line.The cells are bathed in a culture medium, which contains essential nutrients and energy sources necessary for the cells' survival.[16]Thus, in its broader sense, "tissue culture" is often used interchangeably with "cell culture". On the other hand, the strict meaning of "tissue culture" refers to the culturing of tissue pieces, i.e.explant culture.

Tissue culture is an important tool for the study of the biology of cells from multicellular organisms. It provides anin vitromodel of the tissue in a well defined environment which can be easily manipulated and analysed. In animal tissue culture, cells may be grown as two-dimensional monolayers (conventional culture) or within fibrous scaffolds or gels to attain more naturalistic three-dimensional tissue-like structures (3D culture). A 1988 NIH SBIR grant report showed that electrospinning could be used to produce nano- and submicron-scale polymeric fibrous scaffolds specifically intended for use asin vitrocell and tissue substrates. This early use of electrospun fibrous lattices for cell culture and tissue engineering showed that various cell types would adhere to and proliferate upon polycarbonate fibers. It was noted that as opposed to the flattened morphology typically seen in 2D culture, cells grown on the electrospun fibers exhibited a more rounded 3-dimensional morphology generally observed of tissuesin vivo.[17]

Plant tissue culturein particular is concerned with the growing of entire plants from small pieces of plant tissue, cultured in medium.[18]

Concepts in mammalian cell culture

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Isolation of cells

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Main article:Cell isolation

Cells can beisolatedfrom tissues forex vivoculture in several ways. Cells can be easily purified from blood; however, only thewhite cellsare capable of growth in culture. Cells can be isolated from solid tissues by digesting the extracellular matrix usingenzymessuch ascollagenase,trypsin,orpronase,before agitating the tissue to release the cells into suspension.[19][20]Alternatively, pieces of tissue can be placed ingrowth media,and the cells that grow out are available for culture. This method is known asexplant culture.

Cells that are cultured directly from a subject are known as primary cells. With the exception of some derived from tumors, mostprimary cell cultureshave limited lifespan.

An established orimmortalized cell linehas acquired the ability to proliferate indefinitely either through random mutation or deliberate modification, such as artificialexpressionof thetelomerasegene. Numerous cell lines are well established as representative of particularcell types.

Maintaining cells in culture

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For the majority of isolated primary cells, they undergo the process ofsenescenceand stop dividing after a certain number of population doublings while generally retaining their viability (described as theHayflick limit).

A bottle of DMEM cell culture medium

Aside from temperature and gas mixture, the most commonly varied factor in culture systems is the cellgrowth medium.Recipes for growth media can vary inpH,glucose concentration,growth factors,and the presence of other nutrients. The growth factors used to supplement media are often derived from the serum of animal blood, such asfetal bovine serum(FBS), bovine calf serum, equine serum, and porcine serum. One complication of these blood-derived ingredients is the potential for contamination of the culture with viruses orprions,particularly in medicalbiotechnologyapplications. Current practice is to minimize or eliminate the use of these ingredients wherever possible and use humanplatelet lysate(hPL).[21]This eliminates the worry of cross-species contamination when using FBS with human cells. hPL has emerged as a safe and reliable alternative as a direct replacement for FBS or other animal serum. In addition,chemically defined mediacan be used to eliminate any serum trace (human or animal), but this cannot always be accomplished with different cell types. Alternative strategies involve sourcing the animal blood from countries with minimumBSE/TSErisk, such as The United States, Australia and New Zealand,[22]and using purified nutrient concentrates derived from serum in place of whole animal serum for cell culture.[23]

Plating density (number of cells per volume of culture medium) plays a critical role for some cell types. For example, a lower plating density makesgranulosa cellsexhibit estrogen production, while a higher plating density makes them appear asprogesterone-producingtheca lutein cells.[24]

Cells can be grown either insuspensionoradherent cultures.[25]Some cells naturally live in suspension, without being attached to a surface, such as cells that exist in the bloodstream. There are also cell lines that have been modified to be able to survive in suspension cultures so they can be grown to a higher density than adherent conditions would allow. Adherent cells require a surface, such as tissue culture plastic ormicrocarrier,which may be coated with extracellular matrix (such as collagen and laminin) components to increase adhesion properties and provide other signals needed for growth and differentiation. Most cells derived from solid tissues are adherent. Another type of adherent culture isorganotypic culture,which involves growing cells in a three-dimensional (3-D) environment as opposed to two-dimensional culture dishes. This 3D culture system is biochemically and physiologically more similar toin vivotissue, but is technically challenging to maintain because of many factors (e.g. diffusion).[26]

Cell culture basal media

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There are different kinds of cell culture media which being used routinely in life science including the following:

Components of cell culture media

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Component Function
Carbon source (glucose/glutamine) Source of energy
Amino acid Building blocks of protein
Vitamins Promote cell survival and growth
Balanced salt solution Anisotonicmixture of ions to maintain optimumosmotic pressurewithin the cells and provide essential metal ions to act ascofactorsfor enzymatic reactions, cell adhesion etc.
Phenol red dye pH indicator.The color of phenol red changes from orange/red at pH 7–7.4 to yellow at acidic (lower) pH and purple at basic (higher) pH.
Bicarbonate /HEPESbuffer It is used to maintain a balanced pH in the media

Typical Growth conditions

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Parameter
Temperature 37 °C
CO2 5%
Relative Humidity 95%

Cell line cross-contamination

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Main article:List of contaminated cell lines

Cell line cross-contamination can be a problem for scientists working with cultured cells.[27]Studies suggest anywhere from 15 to 20% of the time, cells used in experiments have been misidentified or contaminated with another cell line.[28][29][30]Problems with cell line cross-contamination have even been detected in lines from theNCI-60 panel,which are used routinely for drug-screening studies.[31][32]Major cell line repositories, including theAmerican Type Culture Collection(ATCC), the European Collection of Cell Cultures (ECACC) and the German Collection of Microorganisms and Cell Cultures (DSMZ), have received cell line submissions from researchers that were misidentified by them.[31][33]Such contamination poses a problem for the quality of research produced using cell culture lines, and the major repositories are now authenticating all cell line submissions.[34]ATCC usesshort tandem repeat(STR)DNA fingerprintingto authenticate its cell lines.[35]

To address this problem of cell line cross-contamination, researchers are encouraged to authenticate their cell lines at an early passage to establish the identity of the cell line. Authentication should be repeated before freezing cell line stocks, every two months during active culturing and before any publication of research data generated using the cell lines. Many methods are used to identify cell lines, includingisoenzymeanalysis,human lymphocyte antigen(HLA) typing, chromosomal analysis, karyotyping, morphology andSTR analysis.[35]

One significant cell-line cross contaminant is the immortalHeLacell line. HeLa contamination was first noted in the early 1960s in non-human culture in the USA. Intraspecies contamination was discovered in nineteen cell lines in the seventies. In 1974, five human cell lines from the Soviet Union were found to be HeLa. A follow-up study analysing 50-odd cell lines indicated that half had HeLa markers, but contaminant HeLa had hybridised with the original cell lines. HeLa cell contamination fromair dropletshas been reported. HeLa was even unknowingly injected into human subjects byJonas Salkin a 1978 vaccine trial.[36]

Other technical issues

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As cells generally continue to divide in culture, they generally grow to fill the available area or volume. This can generate several issues:

  • Nutrient depletion in the growth media
  • Changes in pH of the growth media
  • Accumulation ofapoptotic/necrotic(dead) cells
  • Cell-to-cell contact can stimulate cell cycle arrest, causing cells to stop dividing, known ascontact inhibition.
  • Cell-to-cell contact can stimulatecellular differentiation.
  • Geneticandepigeneticalterations, with anatural selectionof the altered cells potentially leading to overgrowth of abnormal, culture-adapted cells with decreased differentiation and increased proliferative capacity.[37]

The choice ofculture mediummight affect thephysiological relevanceof findings from cell culture experiments due to the differences in the nutrient composition and concentrations.[38]A systematic bias in generated datasets was recently shown forCRISPRandRNAigene silencingscreens,[39]and for metabolic profiling of cancercell lines.[38]Using agrowth mediumthat better represents the physiological levels of nutrients can improve the physiological relevance ofin vitrostudies and recently such media types, as Plasmax[40]and Human Plasma Like Medium (HPLM),[41]were developed.

Manipulation of cultured cells

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Among the common manipulations carried out on culture cells are media changes, passaging cells, and transfecting cells. These are generally performed using tissue culture methods that rely onaseptic technique.Aseptic technique aims to avoid contamination with bacteria, yeast, or other cell lines. Manipulations are typically carried out in abiosafety cabinetorlaminar flow cabinetto exclude contaminating micro-organisms.Antibiotics(e.g.penicillinandstreptomycin) and antifungals (e.g.amphotericin BandAntibiotic-Antimycoticsolution) can also be added to the growth media.

As cells undergo metabolic processes, acid is produced and the pH decreases. Often, apH indicatoris added to the medium to measure nutrient depletion.

Media changes

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In the case of adherent cultures, the media can be removed directly by aspiration, and then is replaced. Media changes in non-adherent cultures involve centrifuging the culture and resuspending the cells in fresh media.

Passaging cells

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Main article:Passaging

Passaging (also known as subculture or splitting cells) involves transferring a small number of cells into a new vessel. Cells can be cultured for a longer time if they are split regularly, as it avoids the senescence associated with prolonged high cell density. Suspension cultures are easily passaged with a small amount of culture containing a few cells diluted in a larger volume of fresh media. For adherent cultures, cells first need to be detached; this is commonly done with a mixture oftrypsin-EDTA;however, other enzyme mixes are now available for this purpose. A small number of detached cells can then be used to seed a new culture. Some cell cultures, such asRAW cellsare mechanically scraped from the surface of their vessel with rubber scrapers.

Transfection and transduction

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Main articles:TransfectionandTransformation (genetics)

Another common method for manipulating cells involves the introduction of foreign DNA bytransfection.This is often performed to cause cells toexpress a geneof interest. More recently, the transfection ofRNAiconstructs have been realized as a convenient mechanism for suppressing the expression of a particular gene/protein. DNA can also be inserted into cells using viruses, in methods referred to astransduction,infectionortransformation.Viruses, as parasitic agents, are well suited to introducing DNA into cells, as this is a part of their normal course of reproduction.

Established human cell lines

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CulturedHeLacells have been stained withHoechstturning theirnucleiblue, and are one of the earliest human cell lines descended fromHenrietta Lacks,who died of cervical cancer from which these cells originated.

Cell lines that originate with humans have been somewhat controversial inbioethics,as they may outlive their parent organism and later be used in the discovery of lucrative medical treatments. In the pioneering decision in this area, theSupreme Court of Californiaheld inMoore v. Regents of the University of Californiathat human patients have no property rights in cell lines derived from organs removed with their consent.[42]

Further information:Hybridoma

It is possible to fuse normal cells with animmortalised cell line.This method is used to producemonoclonal antibodies.In brief, lymphocytes isolated from thespleen(or possibly blood) of animmunisedanimal are combined with an immortal myeloma cell line (B cell lineage) to produce ahybridomawhich has the antibody specificity of the primary lymphocyte and the immortality of the myeloma.Selective growth medium(HA or HAT) is used to select against unfused myeloma cells; primary lymphoctyes die quickly in culture and only the fused cells survive. These are screened for production of the required antibody, generally in pools to start with and then after single cloning.

Cell strains

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A cell strain is derived either from a primary culture or a cell line by the selection or cloning of cells having specific properties or characteristics which must be defined. Cell strains are cells that have been adapted to culture but, unlike cell lines, have a finite division potential. Non-immortalized cells stop dividing after 40 to 60 population doublings[43]and, after this, they lose their ability to proliferate (a genetically determined event known as senescence).[44]

Applications of cell culture

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Mass culture of animal cell lines is fundamental to the manufacture of viralvaccinesand other products of biotechnology. Culture of humanstem cellsis used to expand the number of cells and differentiate the cells into various somatic cell types for transplantation.[45]Stem cell culture is also used to harvest the molecules and exosomes that the stem cells release for the purposes of therapeutic development.[46]

Biological products produced byrecombinant DNA(rDNA) technology in animal cell cultures includeenzymes,synthetichormones,immunobiologicals (monoclonal antibodies,interleukins,lymphokines), andanticancer agents.Although many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins that areglycosylated(carbohydrate-modified) currently must be made in animal cells. Mammalian cells ensure expressed proteins are folded correctly and possess human-like glycosylation and post-translational modifications.[47]An important example of such a complex protein is the hormoneerythropoietin.The cost of growing mammalian cell cultures is high, so research is underway to produce such complex proteins in insect cells or in higher plants, use of single embryonic cell andsomaticembryos as a source for direct gene transfer via particle bombardment, transitgene expressionandconfocal microscopyobservation is one of its applications. It also offers to confirm single cell origin of somatic embryos and the asymmetry of the first cell division, which starts the process.

Cell culture is also a key technique forcellular agriculture,which aims to provide both new products and new ways of producing existing agricultural products like milk,(cultured) meat,fragrances, and rhino horn from cells and microorganisms. It is therefore considered one means of achievinganimal-free agriculture.It is also a central tool for teaching cell biology.[48]

Cell culture in two dimensions

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Research intissue engineering,stem cellsandmolecular biologyprimarily involves cultures of cells on flat plastic dishes. This technique is known as two-dimensional (2D) cell culture, and was first developed byWilhelm Rouxwho, in 1885, removed a portion of the medullary plate of an embryonic chicken and maintained it in warm saline for several days on a flat glass plate. From the advance ofpolymertechnology arose today's standard plastic dish for 2D cell culture, commonly known as thePetri dish.Julius Richard Petri,a Germanbacteriologist,is generally credited with this invention while working as an assistant toRobert Koch.Various researchers today also utilize culturinglaboratory flasks,conicals, and even disposable bags like those used insingle-use bioreactors.

Aside from Petri dishes, scientists have long been growing cells within biologically derived matrices such as collagen or fibrin, and more recently, on synthetic hydrogels such as polyacrylamide or PEG. They do this in order to elicit phenotypes that are not expressed on conventionally rigid substrates. There is growing interest in controllingmatrix stiffness,[49]a concept that has led to discoveries in fields such as:

Cell culture in three dimensions

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Cell culture in three dimensionshas been touted as "Biology's New Dimension".[64]At present, the practice of cell culture remains based on varying combinations of single or multiple cell structures in 2D.[65]Currently, there is an increase in use of 3D cell cultures in research areas includingdrug discovery,cancer biology,regenerative medicine,nanomaterialsassessment and basiclife scienceresearch.[66][67][68]3D cell cultures can be grown using a scaffold or matrix, or in a scaffold-free manner. Scaffold based cultures utilize an acellular 3D matrix or a liquid matrix. Scaffold-free methods are normally generated in suspensions.[69]There are a variety of platforms used to facilitate the growth of three-dimensional cellular structures including scaffold systems such as hydrogel matrices[70]and solid scaffolds, and scaffold-free systems such as low-adhesion plates,nanoparticle facilitated magnetic levitation,[71]hanging drop plates,[72][73]androtary cell culture.Culturing cells in 3D leads to wide variation in gene expression signatures and partly mimics tissues in the physiological states.[74]A 3D cell culture model showed cell growth similar to that of in vivo than did a monolayer culture, and all three cultures were capable of sustaining cell growth.[75]As 3D culturing has been developed it turns out to have a great potential to design tumors models and investigate malignant transformation and metastasis, 3D cultures can provide aggerate tool for understanding changes, interactions, and cellular signaling.[76]

3D cell culture in scaffolds

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Eric Simon, in a 1988 NIH SBIR grant report, showed that electrospinning could be used to produce nano- and submicron-scale polystyrene and polycarbonate fibrous scaffolds specifically intended for use asin vitrocell substrates. This early use of electrospun fibrous lattices for cell culture and tissue engineering showed that various cell types including Human Foreskin Fibroblasts (HFF), transformed Human Carcinoma (HEp-2), and Mink Lung Epithelium (MLE) would adhere to and proliferate upon polycarbonate fibers. It was noted that, as opposed to the flattened morphology typically seen in 2D culture, cells grown on the electrospun fibers exhibited a more histotypic rounded 3-dimensional morphology generally observedin vivo.[17]

3D cell culture in hydrogels

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As the naturalextracellular matrix(ECM) is important in the survival, proliferation, differentiation and migration of cells, different hydrogel culture matrices mimicking natural ECM structure are seen as potential approaches to in vivo–like cell culturing.[77]Hydrogels are composed of interconnected pores with high water retention, which enables efficient transport of substances such as nutrients and gases. Several different types of hydrogels from natural and synthetic materials are available for 3D cell culture, including animal ECM extract hydrogels, protein hydrogels, peptide hydrogels, polymer hydrogels, andwood-based nanocellulose hydrogel.

3D Cell Culturing by Magnetic Levitation

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The3D Cell Culturing by Magnetic Levitationmethod (MLM) is the application of growing 3D tissue by inducing cells treated with magnetic nanoparticle assemblies in spatially varying magnetic fields using neodymium magnetic drivers and promoting cell to cell interactions by levitating the cells up to the air/liquid interface of a standard petri dish. The magnetic nanoparticle assemblies consist of magnetic iron oxide nanoparticles, gold nanoparticles, and the polymer polylysine.3D cell culturingis scalable, with the capability for culturing 500 cells to millions of cells or from single dish to high-throughput low volume systems.

Tissue culture and engineering

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Cell culture is a fundamental component oftissue cultureandtissue engineering,as it establishes the basics of growing and maintaining cellsin vitro. The major application of human cell culture is in stem cell industry, wheremesenchymal stem cellscan be cultured and cryopreserved for future use. Tissue engineering potentially offers dramatic improvements in low cost medical care for hundreds of thousands of patients annually.

Vaccines

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Vaccinesforpolio,measles,mumps,rubella,andchickenpoxare currently made in cell cultures. Due to theH5N1pandemicthreat, research into using cell culture forinfluenza vaccinesis being funded by theUnited Statesgovernment. Novel ideas in the field includerecombinant DNA-based vaccines, such as one made using humanadenovirus(a common cold virus) as a vector,[78][79] and novel adjuvants.[80]

Cell co-culture

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The technique of co-culturing is used to study cell crosstalk between two or more types of cells on a plate or in a 3D matrix. The cultivation of different stem cells and the interaction of immune cells can be investigated in an in vitro model similar to biological tissue. Since most tissues contain more than one type of cell, it is important to evaluate their interaction in a 3D culture environment to gain a better understanding of their interaction and to introduce mimetic tissues. There are two types of co-culturing: direct and indirect. While direct interaction involves cells being in direct contact with each other in the same culture media or matrix, indirect interaction involves different environments, allowing signaling and soluble factors to participate.[15][81]

Cell differentiation in tissue models during interaction between cells can be studied using the Co-Cultured System to simulate cancer tumors, to assess the effect of drugs on therapeutic trials, and to study the effect of drugs on therapeutic trials. The co-culture system in 3D models can predict the response to chemotherapy and endocrine therapy if the microenvironment defines biological tissue for the cells.

A co-culture method is used in tissue engineering to generate tissue formation with multiple cells interacting directly.[82]

Schematic representation of 2D culture, 3D culture, organ-on-a-chip and in vivo study

Cell culture in microfluidic device

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Main article:Microfluidic cell culture

Microfluidics technique is developed systems that can perform a process in a flow which are usually in a scale of micron. Microfluidics chip are also known as Lab-on-a-chip and they are able to have continuous procedure and reaction steps with spare amount of reactants and space. Such systems enable the identification and isolation of individual cells and molecules when combined with appropriate biological assays and high-sensitivity detection techniques.[83][84]

Organ-on-a-chip

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Main article:Organ-on-a-chip

OoC systems mimic and control the microenvironment of the cells by growing tissues in microfluidics. Combining tissue engineering, biomaterials fabrication, and cell biology, it offers the possibility of establishing a biomimetic model for studying human diseases in the laboratory. In recent years, 3D cell culture science has made significant progress, leading to the development of OoC. OoC is considered as a preclinical step that benefits pharmaceutical studies,drug developmentand disease modeling.[85][86]OoC is an important technology that can bridge the gap between animal testing and clinical studies and also by the advances that the science has achieved could be a replace for in vivo studies for drug delivery and pathophysiological studies.[87]

Culture of non-mammalian cells

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Besides the culture of well-established immortalised cell lines, cells from primary explants of a plethora of organisms can be cultured for a limited period of time before senescence occurs (see Hayflick's limit). Cultured primary cells have been extensively used in research, as is the case of fish keratocytes in cell migration studies.[88][48][89]

Plant cell culture methods

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Main article:Plant tissue culture
See also:Tobacco BY-2 cells

Plant cell cultures are typically grown as cell suspension cultures in a liquid medium or ascallus cultureson a solid medium. The culturing of undifferentiated plant cells and calli requires the proper balance of the plant growth hormonesauxinandcytokinin.[citation needed]

Insect cell culture

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Main article:Insect cell culture

Cells derived fromDrosophila melanogaster(most prominently,Schneider 2 cells) can be used for experiments which may be hard to do on live flies or larvae, such asbiochemical studiesor studies usingsiRNA.Cell lines derived from the army wormSpodoptera frugiperda,includingSf9andSf21,and from the cabbage looperTrichoplusia ni,High Five cells,are commonly used for expression of recombinant proteins usingbaculovirus.[90]

Bacterial and yeast culture methods

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Main article:Microbiological culture

For bacteria and yeasts, small quantities of cells are usually grown on a solid support that contains nutrients embedded in it, usually a gel such as agar, while large-scale cultures are grown with the cells suspended in a nutrient broth.[citation needed]

Viral culture methods

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Main article:Viral culture

The culture of viruses requires the culture of cells of mammalian, plant, fungal or bacterial origin as hosts for the growth and replication of the virus. Wholewild typeviruses,recombinantviruses or viral products may be generated in cell types other than their natural hosts under the right conditions. Depending on the species of the virus, infection andviral replicationmay result in host cell lysis and formation of aviral plaque.[citation needed]

Common cell lines

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Human cell lines
Animalcell lines
  • Vero(African green monkeyChlorocebuskidneyepithelialcell line)
  • BHK21 cell (Baby Hambster Kidney)
  • MDBK cell (Madin-Darby Bovine Kidney)
  • DF-1cell (chicken fibroblast)
Mousecell lines
Rattumor cell lines
Plant cell lines
Other species cell lines

List of cell lines

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Cell line Meaning Organism Origin tissue Morphology Links
3T3-L1 "3-day transfer, inoculum 3 x 10^5 cells" Mouse Embryo Fibroblast ECACCCellosaurus
4T1 Mouse Mammary gland ATCCCellosaurus
1321N1 Human Brain Astrocytoma ECACCCellosaurus
9L Rat Brain Glioblastoma ECACCCellosaurus
A172 Human Brain Glioblastoma ECACCCellosaurus
A20 Mouse Blymphoma Blymphocyte Cellosaurus
A253 Human Submandibular duct Head and neck carcinoma ATCCCellosaurus
A2780 Human Ovary Ovarian carcinoma ECACCCellosaurus
A2780ADR Human Ovary Adriamycin-resistant derivative of A2780 ECACCCellosaurus
A2780cis Human Ovary Cisplatin-resistant derivative of A2780 ECACCCellosaurus
A431 Human Skin epithelium Squamous cell carcinoma ECACCCellosaurus
A549 Human Lung Lung carcinoma ECACCCellosaurus
AB9 Zebrafish Fin Fibroblast ATCCCellosaurus
AHL-1 Armenian Hamster Lung-1 Hamster Lung ECACCArchived24 November 2021 at theWayback MachineCellosaurus
ALC Mouse Bone marrow Stroma PMID2435412[91]Cellosaurus
B16 Mouse Melanoma ECACCArchived24 November 2021 at theWayback MachineCellosaurus
B35 Rat Neuroblastoma ATCCCellosaurus
BCP-1 Human PBMC HIV+ primary effusion lymphoma ATCCCellosaurus
BEAS-2B Bronchial epithelium + Adenovirus 12-SV40 virus hybrid (Ad12SV40) Human Lung Epithelial ECACCCellosaurus
bEnd.3 Brain Endothelial 3 Mouse Brain/cerebral cortex Endothelium Cellosaurus
BHK-21 Baby Hamster Kidney-21 Hamster Kidney Fibroblast ECACCArchived24 November 2021 at theWayback MachineCellosaurus
BOSC23 Packaging cell line derived fromHEK 293 Human Kidney (embryonic) Epithelium Cellosaurus
BT-20 Breast Tumor-20 Human Breast epithelium Breast carcinoma ATCCCellosaurus
BxPC-3 Biopsy xenograft of Pancreatic Carcinoma line 3 Human Pancreatic adenocarcinoma Epithelial ECACCCellosaurus
C2C12 Mouse Myoblast ECACCCellosaurus
C3H-10T1/2 Mouse Embryonic mesenchymal cell line ECACCCellosaurus
C6 Rat Brainastrocyte Glioma ECACCCellosaurus
C6/36 Insect -Asian tiger mosquito Larval tissue ECACCCellosaurus
Caco-2 Human Colon Colorectal carcinoma ECACCCellosaurus
Cal-27 Human Tongue Squamous cell carcinoma ATCCCellosaurus
Calu-3 Human Lung Adenocarcinoma ATCCCellosaurus
CGR8 Mouse Embryonic stem cells ECACCCellosaurus
CHO Chinese Hamster Ovary Hamster Ovary Epithelium ECACCArchived29 October 2021 at theWayback MachineCellosaurus
CML T1 Chronic myeloid leukemia T lymphocyte 1 Human CML acute phase T cell leukemia DSMZCellosaurus
CMT12 Canine Mammary Tumor 12 Dog Mammary gland Epithelium Cellosaurus
COR-L23 Human Lung Lung carcinoma ECACCCellosaurus
COR-L23/5010 Human Lung Lung carcinoma ECACCCellosaurus
COR-L23/CPR Human Lung Lung carcinoma ECACCCellosaurus
COR-L23/R23- Human Lung Lung carcinoma ECACCCellosaurus
COS-7 Cercopithecus aethiops,origin-defective SV-40 Old World monkey -Cercopithecus aethiops(Chlorocebus) Kidney Fibroblast ECACCCellosaurus
COV-434 Human Ovary Ovarian granulosa cell carcinoma PMID8436435[92]ECACCCellosaurus
CT26 Mouse Colon Colorectal carcinoma Cellosaurus
D17 Dog Lung metastasis Osteosarcoma ATCCCellosaurus
DAOY Human Brain Medulloblastoma ATCCCellosaurus
DH82 Dog Histiocytosis Monocyte/macrophage ECACCCellosaurus
DU145 Human Androgeninsensitive prostate carcinoma ATCCCellosaurus
DuCaP Dura matercancer of the Prostate Human Metastatic prostate carcinoma Epithelial PMID11317521[93]Cellosaurus
E14Tg2a Mouse Embryonic stem cells ECACCCellosaurus
EL4 Mouse T cell leukemia ECACCCellosaurus
EM-2 Human CML blast crisis Ph+ CML line DSMZCellosaurus
EM-3 Human CML blast crisis Ph+ CML line DSMZCellosaurus
EMT6/AR1 Mouse Mammary gland Epithelial-like ECACCCellosaurus
EMT6/AR10.0 Mouse Mammary gland Epithelial-like ECACCCellosaurus
FM3 Human Lymph node metastasis Melanoma ECACCCellosaurus
GL261 Glioma 261 Mouse Brain Glioma Cellosaurus
H1299 Human Lung Lung carcinoma ATCCCellosaurus
HaCaT Human Skin Keratinocyte CLSCellosaurus
HCA2 Human Colon Adenocarcinoma ECACCCellosaurus
HEK 293 Human Embryonic Kidney 293 Human Kidney (embryonic) Epithelium ECACCCellosaurus
HEK 293T HEK 293derivative Human Kidney (embryonic) Epithelium ECACCCellosaurus
HeLa "Henrietta Lacks" Human Cervix epithelium Cervical carcinoma ECACCCellosaurus
Hepa1c1c7 Clone 7 of clone 1 hepatoma line 1 Mouse Hepatoma Epithelial ECACCCellosaurus
Hep G2 Human Liver Hepatoblastoma ECACCCellosaurus
High Five Insect (moth) -Trichoplusia ni Ovary Cellosaurus
HL-60 Human Leukemia-60 Human Blood Myeloblast ECACCCellosaurus
HT-1080 Human Fibrosarcoma ECACCCellosaurus
HT-29 Human Colon epithelium Adenocarcinoma ECACCCellosaurus
J558L Mouse Myeloma B lymphocyte cell ECACCCellosaurus
Jurkat Human White blood cells T cellleukemia ECACCCellosaurus
JY Human Lymphoblastoid EBV-transformed B cell ECACCCellosaurus
K562 Human Lymphoblastoid CML blast crisis ECACCCellosaurus
KBM-7 Human Lymphoblastoid CML blast crisis Cellosaurus
KCL-22 Human Lymphoblastoid CML DSMZCellosaurus
KG1 Human Lymphoblastoid AML ECACCCellosaurus
Ku812 Human Lymphoblastoid Erythroleukemia ECACCCellosaurus
KYO-1 Kyoto-1 Human Lymphoblastoid CML DSMZCellosaurus
L1210 Mouse Lymphocytic leukemia Ascitic fluid ECACCCellosaurus
L243 Mouse Hybridoma Secretes L243 mAb (against HLA-DR) ATCCCellosaurus
LNCaP Lymph Node Cancer of the Prostate Human Prostatic adenocarcinoma Epithelial ECACCCellosaurus
MA-104 Microbiological Associates-104 African Green Monkey Kidney Epithelial Cellosaurus
MA2.1 Mouse Hybridoma Secretes MA2.1 mAb (against HLA-A2 and HLA-B17) ATCCCellosaurus
Ma-Mel 1, 2, 3....48 Human Skin A range ofmelanomacell lines ECACCArchived24 November 2021 at theWayback MachineCellosaurus
MC-38 Mouse Colon-38 Mouse Colon Adenocarcinoma Cellosaurus
MCF-7 Michigan Cancer Foundation-7 Human Breast Invasive breast ductal carcinoma ER+, PR+ ECACCCellosaurus
MCF-10A Michigan Cancer Foundation-10A Human Breast epithelium ATCCCellosaurus
MDA-MB-157 M.D. Anderson - Metastatic Breast-157 Human Pleural effusion metastasis Breast carcinoma ECACCCellosaurus
MDA-MB-231 M.D. Anderson - Metastatic Breast-231 Human Pleural effusion metastasis Breast carcinoma ECACCCellosaurus
MDA-MB-361 M.D. Anderson - Metastatic Breast-361 Human Melanoma(contaminated by M14) ECACCCellosaurus
MDA-MB-468 M.D. Anderson - Metastatic Breast-468 Human Pleural effusion metastasis Breast carcinoma ATCCCellosaurus
MDCK II Madin Darby Canine Kidney II Dog Kidney Epithelium ECACCCellosaurus
MG63 Human Bone Osteosarcoma ECACCCellosaurus
MIA PaCa-2 Human Prostate Pancreatic Carcinoma ATCCCellosaurus
MOR/0.2R Human Lung Lung carcinoma ECACCCellosaurus
Mono-Mac-6 Human White blood cells Myeloid metaplasicAML DSMZCellosaurus
MRC-5 Medical Research Council cell strain 5 Human Lung (fetal) Fibroblast ECACCArchived24 November 2021 at theWayback MachineCellosaurus
MTD-1A Mouse Epithelium Cellosaurus
MyEnd Myocardial Endothelial Mouse Endothelium Cellosaurus
NCI-H69 Human Lung Lung carcinoma ECACCCellosaurus
NCI-H69/CPR Human Lung Lung carcinoma ECACCCellosaurus
NCI-H69/LX10 Human Lung Lung carcinoma ECACCCellosaurus
NCI-H69/LX20 Human Lung Lung carcinoma ECACCCellosaurus
NCI-H69/LX4 Human Lung Lung carcinoma ECACCCellosaurus
Neuro-2a Mouse Nerve/neuroblastoma Neuronal stem cells ECACCCellosaurus
NIH-3T3 NIH,3-day transfer, inoculum 3 x 105cells Mouse Embryo Fibroblast ECACCCellosaurus
NALM-1 Human Peripheral blood Blast-crisis CML ATCCCellosaurus
NK-92 Human Leukemia/lymphoma ATCCCellosaurus
NTERA-2 Human Lung metastasis Embryonal carcinoma ECACCCellosaurus
NW-145 Human Skin Melanoma ESTDABArchived2011-11-16 at theWayback MachineCellosaurus
OK Opossum Kidney Virginia opossum-Didelphis virginiana Kidney ECACCCellosaurus
OPCN / OPCT cell lines Human Prostate Range of prostate tumour lines Cellosaurus
P3X63Ag8 Mouse Myeloma ECACCCellosaurus
PANC-1 Human Duct Epithelioid Carcinoma ATCCCellosaurus
PC12 Rat Adrenal medulla Pheochromocytoma ECACCCellosaurus
PC-3 Prostate Cancer-3 Human Bone metastasis Prostate carcinoma ECACCCellosaurus
Peer Human T cell leukemia DSMZCellosaurus
PNT1A Human Prostate SV40-transformed tumour line ECACCCellosaurus
PNT2 Human Prostate SV40-transformed tumour line ECACCCellosaurus
Pt K2 The second cell line derived fromPotorous tridactylis Long-nosed potoroo-Potorous tridactylus Kidney Epithelial ECACCCellosaurus
Raji Human Blymphoma Lymphoblast-like ECACCCellosaurus
RBL-1 Rat Basophilic Leukemia-1 Rat Leukemia Basophil cell ECACCCellosaurus
RenCa Renal Carcinoma Mouse Kidney Renal carcinoma ATCCCellosaurus
RIN-5F Mouse Pancreas ECACCCellosaurus
RMA-S Mouse T cell tumour Cellosaurus
S2 Schneider 2 Insect -Drosophila melanogaster Late stage (20–24 hours old) embryos ATCCCellosaurus
SaOS-2 Sarcoma OSteogenic-2 Human Bone Osteosarcoma ECACCCellosaurus
Sf21 Spodoptera frugiperda21 Insect (moth) -Spodoptera frugiperda Ovary ECACCCellosaurus
Sf9 Spodoptera frugiperda9 Insect (moth) -Spodoptera frugiperda Ovary ECACCCellosaurus
SH-SY5Y Human Bone marrow metastasis Neuroblastoma ECACCCellosaurus
SiHa Human Cervix epithelium Cervical carcinoma ATCCCellosaurus
SK-BR-3 Sloan-KetteringBreast cancer 3 Human Breast Breast carcinoma DSMZCellosaurus
SK-OV-3 Sloan-KetteringOvarian cancer 3 Human Ovary Ovarian carcinoma ECACCCellosaurus
SK-N-SH Human Brain Epithelial ATCCCellosaurus
T2 Human T cell leukemia/B cell linehybridoma ATCCCellosaurus
T-47D Human Breast Breast ductal carcinoma ECACCCellosaurus
T84 Human Lung metastasis Colorectal carcinoma ECACCCellosaurus
T98G Human Glioblastoma-astrocytoma Epithelium ECACCCellosaurus
THP-1 Human Monocyte Acute monocytic leukemia ECACCCellosaurus
U2OS Human Osteosarcoma Epithelial ECACCCellosaurus
U373 Human Glioblastoma-astrocytoma Epithelium ECACCArchived24 November 2021 at theWayback MachineCellosaurus
U87 Human Glioblastoma-astrocytoma Epithelial-like ECACCCellosaurus
U937 Human Leukemic monocytic lymphoma ECACCCellosaurus
VCaP Vertebral Cancer of the Prostate Human Vertebra metastasis Prostate carcinoma ECACCCellosaurus
Vero From Esperanto:verda(green, for green monkey)reno(kidney) African green monkey -Chlorocebus sabaeus Kidney epithelium ECACCCellosaurus
VG-1 Human Primary effusion lymphoma Cellosaurus
WM39 Human Skin Melanoma ESTDABCellosaurus
WT-49 Human Lymphoblastoid ECACCCellosaurus
YAC-1 Mouse Lymphoma ECACCCellosaurus
YAR Human Lymphoblastoid EBV-transformed B cell Human Immunology[94]ECACCCellosaurus

See also

edit

References and notes

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