Cell culture






Cell culture in a special tissue culture dish





Epithelial cells in culture, stained for keratin (red) and DNA (green)


Cell culture is the process by which cells are grown under controlled conditions, generally outside their natural environment. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or medium that 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 (adherent or monolayer culture) whereas others can be grown free floating in culture medium (suspension culture). 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.


In practice, the term "cell culture" now refers to the culturing of cells derived from multicellular eukaryotes, especially animal cells, in contrast with other types of culture that also grow cells, such as plant tissue culture, fungal culture, and microbiological culture (of microbes). The historical development and methods of cell culture are closely interrelated to those of tissue culture and organ culture. Viral culture is also related, with cells as hosts for the viruses.


The laboratory technique of maintaining live cell 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.[1][2]




Contents






  • 1 History


  • 2 Concepts in mammalian cell culture


    • 2.1 Isolation of cells


    • 2.2 Maintaining cells in culture


    • 2.3 Components of cell culture media


      • 2.3.1 Typical Growth conditions




    • 2.4 Cell line cross-contamination


    • 2.5 Other technical issues


    • 2.6 Manipulation of cultured cells


      • 2.6.1 Media changes


      • 2.6.2 Passaging cells


      • 2.6.3 Transfection and transduction




    • 2.7 Established human cell lines


    • 2.8 Cell strains




  • 3 Applications of cell culture


    • 3.1 Cell culture in two dimensions


    • 3.2 Cell culture in three dimensions


      • 3.2.1 3D cell culture in hydrogels


      • 3.2.2 3D Cell Culturing by Magnetic Levitation




    • 3.3 Tissue culture and engineering


    • 3.4 Vaccines




  • 4 Culture of non-mammalian cells


    • 4.1 Plant cell culture methods


    • 4.2 Insect cell culture


    • 4.3 Bacterial and yeast culture methods


    • 4.4 Viral culture methods




  • 5 Common cell lines


  • 6 List of cell lines


  • 7 See also


  • 8 References and notes


  • 9 Further reading


  • 10 External links





History


The 19th-century English physiologist Sydney Ringer developed salt solutions containing the chlorides of sodium, potassium, calcium and magnesium suitable for maintaining the beating of an isolated animal heart outside the body.[3] In 1885, Wilhelm Roux removed a portion of the medullary plate of an embryonic chicken and maintained it in a warm saline solution for several days, establishing the principle of tissue culture.[4]Ross Granville Harrison, working at Johns Hopkins Medical School and then at Yale University, published results of his experiments from 1907 to 1910, establishing the methodology of tissue culture.[5]


Cell culture techniques were advanced significantly in the 1940s and 1950s to support research in virology. Growing viruses in cell cultures allowed preparation of purified viruses for the manufacture of vaccines. The injectable polio vaccine developed by Jonas Salk was one of the first products mass-produced using cell culture techniques. This vaccine was made possible by the cell culture research of John Franklin Enders, Thomas Huckle Weller, and Frederick Chapman Robbins, who were awarded a Nobel Prize for their discovery of a method of growing the virus in monkey kidney cell cultures.



Concepts in mammalian cell culture



Isolation of cells


Main article: Cell isolation

Cells can be isolated from tissues for ex vivo culture in several ways. Cells can be easily purified from blood; however, only the white cells are capable of growth in culture. Cells can be isolated from solid tissues by digesting the extracellular matrix using enzymes such as collagenase, trypsin, or pronase, before agitating the tissue to release the cells into suspension.[6][7] Alternatively, pieces of tissue can be placed in growth media, and the cells that grow out are available for culture. This method is known as explant culture.


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


An established or immortalized cell line has acquired the ability to proliferate indefinitely either through random mutation or deliberate modification, such as artificial expression of the telomerase gene.
Numerous cell lines are well established as representative of particular cell types.



Maintaining cells in culture


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



Cells are grown and maintained at an appropriate temperature and gas mixture (typically, 37 °C, 5% CO2 for mammalian cells) in a cell incubator. Culture conditions vary widely for each cell type, and variation of conditions for a particular cell type can result in different phenotypes.




A bottle of DMEM cell culture medium


Aside from temperature and gas mixture, the most commonly varied factor in culture systems is the cell growth medium. Recipes for growth media can vary in pH, 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 as fetal 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 or prions, particularly in medical biotechnology applications. Current practice is to minimize or eliminate the use of these ingredients wherever possible and use human platelet lysate (hPL).[8] 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 media can 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 minimum BSE/TSE risk, such as The United States, Australia and New Zealand,[9] and using purified nutrient concentrates derived from serum in place of whole animal serum for cell culture.[10]


Plating density (number of cells per volume of culture medium) plays a critical role for some cell types. For example, a lower plating density makes granulosa cells exhibit estrogen production, while a higher plating density makes them appear as progesterone-producing theca lutein cells.[11]


Cells can be grown either in suspension or adherent cultures. 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 or microcarrier, 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 is organotypic 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 to in vivo tissue, but is technically challenging to maintain because of many factors (e.g. diffusion).



Components of cell culture media































Component
 Function
Carbon source (glucose/glutamine)
 Source of energy

Amino acid
 Building blocks of protein

Vitamins
 Promote cell survival and growth
Balanced salt solution
 An isotonic mixture of ions to maintain optimum osmotic pressure within the cells and provide essential metal ions to act as cofactors for 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 /HEPES buffer
 It is used to maintain a balanced pH in the media


Typical Growth conditions



















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


Cell line cross-contamination


Main article: List of contaminated cell lines

Cell line cross-contamination can be a problem for scientists working with cultured cells.[12] Studies suggest anywhere from 15–20% of the time, cells used in experiments have been misidentified or contaminated with another cell line.[13][14][15] Problems with cell line cross-contamination have even been detected in lines from the NCI-60 panel, which are used routinely for drug-screening studies.[16][17] Major cell line repositories, including the American 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.[16][18] 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.[19] ATCC uses short tandem repeat (STR) DNA fingerprinting to authenticate its cell lines.[20]


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, including isoenzyme analysis, human lymphocyte antigen (HLA) typing, chromosomal analysis, karyotyping, morphology and STR analysis.[20]


One significant cell-line cross contaminant is the immortal HeLa cell line.



Other technical issues


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 of apoptotic/necrotic (dead) cells

  • Cell-to-cell contact can stimulate cell cycle arrest, causing cells to stop dividing, known as contact inhibition.

  • Cell-to-cell contact can stimulate cellular differentiation.


  • Genetic and epigenetic alterations, with a natural selection of the altered cells potentially leading to overgrowth of abnormal, culture-adapted cells with decreased differentiation and increased proliferative capacity.[21]



Manipulation of cultured cells


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 on aseptic technique. Aseptic technique aims to avoid contamination with bacteria, yeast, or other cell lines. Manipulations are typically carried out in a biosafety hood or laminar flow cabinet to exclude contaminating micro-organisms. Antibiotics (e.g. penicillin and streptomycin) and antifungals (e.g.amphotericin B) can also be added to the growth media.


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



Media changes


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


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 of trypsin-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 as RAW cells are mechanically scraped from the surface of their vessel with rubber scrapers.



Transfection and transduction


Main articles: Transfection and Transformation (genetics)

Another common method for manipulating cells involves the introduction of foreign DNA by transfection. This is often performed to cause cells to express a gene of interest. More recently, the transfection of RNAi constructs 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 as transduction, infection or transformation. 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




Cultured HeLa cells have been stained with Hoechst turning their nuclei blue, and are one of the earliest human cell lines descended from Henrietta Lacks, who died of cervical cancer from which these cells originated.


Cell lines that originate with humans have been somewhat controversial in bioethics, 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, the Supreme Court of California held in Moore v. Regents of the University of California that human patients have no property rights in cell lines derived from organs removed with their consent.[22]


Further information: Hybridoma

It is possible to fuse normal cells with an immortalised cell line. This method is used to produce monoclonal antibodies. In brief, lymphocytes isolated from the spleen (or possibly blood) of an immunised animal are combined with an immortal myeloma cell line (B cell lineage) to produce a hybridoma which 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


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[23] and, after this, they lose their ability to proliferate (a genetically determined event known as senescence).[24]



Applications of cell culture


Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and other products of biotechnology. Culture of human stem cells is used to expand the number of cells and differentiate the cells into various somatic cell types for transplantation.[25] Stem cell culture is also used to harvest the molecules and exosomes that the stem cells release for the purposes of therapeutic development.[26]


Biological products produced by recombinant DNA (rDNA) technology in animal cell cultures include enzymes, synthetic hormones, immunobiologicals (monoclonal antibodies, interleukins, lymphokines), and anticancer agents. Although many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins that are glycosylated (carbohydrate-modified) currently must be made in animal cells. An important example of such a complex protein is the hormone erythropoietin. 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 and somatic embryos as a source for direct gene transfer via particle bombardment, transit gene expression and confocal microscopy observation 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 for cellular 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 achieving animal-free agriculture. It is also a central tool for teaching cell biology.[27]



Cell culture in two dimensions


Research in tissue engineering, stem cells and molecular biology primarily involves cultures of cells on flat plastic dishes. This technique is known as two-dimensional (2D) cell culture, and was first developed by Wilhelm Roux who, 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 of polymer technology arose today's standard plastic dish for 2D cell culture, commonly known as the Petri dish. Julius Richard Petri, a German bacteriologist, is generally credited with this invention while working as an assistant to Robert Koch. Various researchers today also utilize culturing laboratory flasks, conicals, and even disposable bags like those used in single-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 controlling matrix stiffness,[28] a concept that has led to discoveries in fields such as:



  • Stem cell self-renewal[29][30]

  • Lineage specification[31]

  • Cancer cell phenotype[32][33][34]

  • Fibrosis[35][36]

  • Hepatocyte function[37][38][39]

  • Mechanosensing[40][41][42]



Cell culture in three dimensions


Cell culture in three dimensions has been touted as "Biology's New Dimension".[43] At present, the practice of cell culture remains based on varying combinations of single or multiple cell structures in 2D.[44] Currently, there is an increase in use of 3D cell cultures in research areas including drug discovery, cancer biology, regenerative medicine and basic life science research.[45] 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.[46] There are a variety of platforms used to facilitate the growth of three-dimensional cellular structures including scaffold systems such as hydrogel matrices[47] and solid scaffolds, and scaffold-free systems such as low-adhesion plates, nanoparticle facilitated magnetic levitation,[48] and hanging drop plates.[49]


3D cell culture in scaffolds


Eric Simon, in a 1988 NIH SBIR grant report, showed that electrospinning could be used to produced nano- and submicron-scale polystyrene and polycarbonate fibrous scaffolds specifically intended for use as in vitro cell 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 observed in vivo.[50]



3D cell culture in hydrogels


As the natural extracellular 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.[51] 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, and wood-based nanocellulose hydrogel.



3D Cell Culturing by Magnetic Levitation


The 3D Cell Culturing by Magnetic Levitation method (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 culturing is 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


Cell culture is a fundamental component of tissue culture and tissue engineering, as it establishes the basics of growing and maintaining cells in vitro.
The major application of human cell culture is in stem cell industry, where mesenchymal stem cells can 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


Vaccines for polio, measles, mumps, rubella, and chickenpox are currently made in cell cultures. Due to the H5N1 pandemic threat, research into using cell culture for influenza vaccines is being funded by the United States government. Novel ideas in the field include recombinant DNA-based vaccines, such as one made using human adenovirus (a common cold virus) as a vector,[52][53]
and novel adjuvants.[54]



Culture of non-mammalian cells


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 sensecence 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.[55][27][56]



Plant cell culture methods


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 as callus cultures on a solid medium. The culturing of undifferentiated plant cells and calli requires the proper balance of the plant growth hormones auxin and cytokinin.



Insect cell culture


Cells derived from Drosophila melanogaster (most prominently, Schneider 2 cells) can be used for experiments which may be hard to do on live flies or larvae, such as biochemical studies or studies using siRNA. Cell lines derived from the army worm Spodoptera frugiperda, including Sf9 and Sf21, and from the cabbage looper Trichoplusia ni, High Five cells, are commonly used for expression of recombinant proteins using baculovirus.



Bacterial and yeast culture methods


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.



Viral culture methods


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. Whole wild type viruses, recombinant viruses 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 and viral replication may result in host cell lysis and formation of a viral plaque.



Common cell lines


Human cell lines



  • DU145 (prostate cancer)


  • H295R (adrenocortical cancer)


  • HeLa (cervical cancer)


  • KBM-7 (chronic myelogenous leukemia)


  • LNCaP (prostate cancer)


  • MCF-7 (breast cancer)


  • MDA-MB-468 (breast cancer)


  • PC3 (prostate cancer)


  • SaOS-2 (bone cancer)


  • SH-SY5Y (neuroblastoma, cloned from a myeloma)


  • T-47D (breast cancer)


  • THP-1 (acute myeloid leukemia)


  • U87 (glioblastoma)


  • National Cancer Institute's 60 cancer cell line panel (NCI60)



Primate cell lines


  • Vero (African green monkey Chlorocebus kidney epithelial cell line)


Mouse cell lines


  • MC3T3 (embryonic calvarium)


Rat tumor cell lines


  • GH3 (pituitary tumor)


  • PC12 (pheochromocytoma)


Plant cell lines


  • Tobacco BY-2 cells (kept as cell suspension culture, they are model system of plant cell)

Other species cell lines



  • Dog MDCK kidney epithelial


  • Xenopus A6 kidney epithelial


  • Zebrafish AB9



List of cell lines


This list is incomplete; you can help by expanding it.










































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































Cell line Meaning Organism Origin tissue Morphology Links
3T3-L1 "3-day transfer, inoculum 3 x 10^5 cells" Mouse Embryo Fibroblast
ECACC Cellosaurus
4T1 Mouse Mammary gland
ATCC Cellosaurus
9L Rat Brain Glioblastoma
ECACC Cellosaurus
A172 Human Brain Glioblastoma
ECACC Cellosaurus
A20 Mouse B lymphoma
 B lymphocyte
Cellosaurus
A253 Human Submandibular duct Head and neck carcinoma
ATCC Cellosaurus
A2780 Human Ovary Ovarian carcinoma
ECACC Cellosaurus
A2780ADR Human Ovary Adriamycin-resistant derivative of A2780
ECACC Cellosaurus
A2780cis Human Ovary Cisplatin-resistant derivative of A2780
ECACC Cellosaurus
A431 Human Skin epithelium Squamous cell carcinoma
ECACC Cellosaurus
A549 Human Lung Lung carcinoma
ECACC Cellosaurus
AB9 Zebrafish Fin Fibroblast
ATCC Cellosaurus
AHL-1 Armenian Hamster Lung-1 Hamster Lung
ECACC Cellosaurus
ALC Mouse Bone marrow Stroma
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PMID 2435412[57]Cellosaurus
B16 Mouse Melanoma
ECACC Cellosaurus
B35 Rat Neuroblastoma
ATCC Cellosaurus
BCP-1 Human PBMC HIV+ primary effusion lymphoma
ATCC Cellosaurus
BEAS-2B Bronchial epithelium + Adenovirus 12-SV40 virus hybrid (Ad12SV40) Human Lung Epithelial
ECACC Cellosaurus
bEnd.3 Brain Endothelial 3 Mouse Brain/cerebral cortex
 Endothelium
Cellosaurus
BHK-21 Baby Hamster Kidney-21 Hamster Kidney Fibroblast
ECACC Cellosaurus
BOSC23 Packaging cell line derived from HEK 293
 Human Kidney (embryonic) Epithelium
Cellosaurus
BT-20 Breast Tumor-20 Human Breast epithelium Breast carcinoma
ATCC Cellosaurus
BxPC-3 Biopsy xenograft of Pancreatic Carcinoma line 3 Human Pancreatic adenocarcinoma Epithelial
ECACC Cellosaurus
C2C12 Mouse Myoblast
ECACC Cellosaurus
C3H-10T1/2 Mouse Embryonic mesenchymal cell line
ECACC Cellosaurus
C6 Rat Brain astrocyte
 Glioma
ECACC Cellosaurus
C6/36 Insect - Asian tiger mosquito
 Larval tissue
ECACC Cellosaurus
Caco-2 Human Colon Colorectal carcinoma
ECACC Cellosaurus
Cal-27 Human Tongue Squamous cell carcinoma
ATCC Cellosaurus
Calu-3 Human Lung Adenocarcinoma
ATCC Cellosaurus
CGR8 Mouse Embryonic stem cells
ECACC Cellosaurus
CHO Chinese Hamster Ovary Hamster Ovary Epithelium
ECACC Cellosaurus
CML T1 Chronic myeloid leukemia T lymphocyte 1 Human CML acute phase T cell leukemia
DSMZ Cellosaurus
CMT12 Canine Mammary Tumor 12 Dog Mammary gland Epithelium
Cellosaurus
COR-L23 Human Lung Lung carcinoma
ECACC Cellosaurus
COR-L23/5010 Human Lung Lung carcinoma
ECACC Cellosaurus
COR-L23/CPR Human Lung Lung carcinoma
ECACC Cellosaurus
COR-L23/R23- Human Lung Lung carcinoma
ECACC Cellosaurus
COS-7
Cercopithecus aethiops, origin-defective SV-40		 Old World monkey - Cercopithecus aethiops (Chlorocebus) Kidney Fibroblast
ECACC Cellosaurus
COV-434 Human Ovary Ovarian granulosa cell carcinoma

PMID 8436435[58]ECACC Cellosaurus
CT26 Mouse Colon Colorectal carcinoma
Cellosaurus
D17 Dog Lung metastasis Osteosarcoma
ATCC Cellosaurus
DAOY Human Brain Medulloblastoma
ATCC Cellosaurus
DH82 Dog Histiocytosis
Monocyte/macrophage
ECACC Cellosaurus
DU145 Human
Androgen insensitive prostate carcinoma
ATCC Cellosaurus
DuCaP
Dura mater cancer of the Prostate		 Human Metastatic prostate carcinoma Epithelial

PMID 11317521[59]Cellosaurus
E14Tg2a Mouse Embryonic stem cells
ECACC Cellosaurus
EL4 Mouse T cell leukemia
ECACC Cellosaurus
EM-2 Human CML blast crisis Ph+ CML line
DSMZ Cellosaurus
EM-3 Human CML blast crisis Ph+ CML line
DSMZ Cellosaurus
EMT6/AR1 Mouse Mammary gland Epithelial-like
ECACC Cellosaurus
EMT6/AR10.0 Mouse Mammary gland Epithelial-like
ECACC Cellosaurus
FM3 Human Lymph node metastasis Melanoma
ECACC Cellosaurus
GL261 Glioma 261 Mouse Brain Glioma
Cellosaurus
H1299 Human Lung Lung carcinoma
ATCC Cellosaurus
HaCaT Human Skin Keratinocyte
CLS Cellosaurus
HCA2 Human Colon Adenocarcinoma
ECACC Cellosaurus
HEK 293 Human Embryonic Kidney 293 Human Kidney (embryonic) Epithelium
ECACC Cellosaurus
HEK 293T
HEK 293 derivative		 Human Kidney (embryonic) Epithelium
ECACC Cellosaurus
HeLa "Henrietta Lacks" Human Cervix epithelium Cervical carcinoma
ECACC Cellosaurus
Hepa1c1c7 Clone 7 of clone 1 hepatoma line 1 Mouse Hepatoma Epithelial
ECACC Cellosaurus
Hep G2 Human Liver Hepatoblastoma
ECACC Cellosaurus
High Five Insect (moth) - Trichoplusia ni
 Ovary
Cellosaurus
HL-60 Human Leukemia-60 Human Blood Myeloblast
ECACC Cellosaurus
HT-1080 Human Fibrosarcoma
ECACC Cellosaurus
HT-29 Human Colon epithelium Adenocarcinoma
ECACC Cellosaurus
J558L Mouse Myeloma B lymphocyte cell
ECACC Cellosaurus
Jurkat Human White blood cells T cell leukemia
ECACC Cellosaurus
JY Human Lymphoblastoid EBV-transformed B cell
ECACC Cellosaurus
K562 Human Lymphoblastoid CML blast crisis
ECACC Cellosaurus
KBM-7 Human Lymphoblastoid CML blast crisis
Cellosaurus
KCL-22 Human Lymphoblastoid CML
DSMZ Cellosaurus
KG1 Human Lymphoblastoid AML
ECACC Cellosaurus
Ku812 Human Lymphoblastoid Erythroleukemia
ECACC Cellosaurus
KYO-1 Kyoto-1 Human Lymphoblastoid CML
DSMZ Cellosaurus
L1210 Mouse Lymphocytic leukemia Ascitic fluid
ECACC Cellosaurus
L243 Mouse Hybridoma Secretes L243 mAb (against HLA-DR)
ATCC Cellosaurus
LNCaP Lymph Node Cancer of the Prostate Human Prostatic adenocarcinoma Epithelial
ECACC Cellosaurus
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)
ATCC Cellosaurus
Ma-Mel 1, 2, 3....48 Human Skin A range of melanoma cell lines
ECACC Cellosaurus
MC-38 Mouse Colon-38 Mouse Colon Adenocarcinoma
Cellosaurus
MCF-7 Michigan Cancer Foundation-7 Human Breast Invasive breast ductal carcinoma ER+, PR+
ECACC Cellosaurus
MCF-10A Michigan Cancer Foundation-10A Human Breast epithelium
ATCC Cellosaurus
MDA-MB-157 M.D. Anderson - Metastatic Breast-157 Human Pleural effusion metastasis Breast carcinoma
ECACC Cellosaurus
MDA-MB-231 M.D. Anderson - Metastatic Breast-231 Human Pleural effusion metastasis Breast carcinoma
ECACC Cellosaurus
MDA-MB-361 M.D. Anderson - Metastatic Breast-361 Human
Melanoma (contaminated by M14)
ECACC Cellosaurus
MDA-MB-468 M.D. Anderson - Metastatic Breast-468 Human Pleural effusion metastasis Breast carcinoma
ATCC Cellosaurus
MDCK II Madin Darby Canine Kidney II Dog Kidney Epithelium
ECACC Cellosaurus
MG63 Human Bone Osteosarcoma
ECACC Cellosaurus
MIA PaCa-2 Human Prostate Pancreatic Carcinoma
ATCC Cellosaurus
MOR/0.2R Human Lung Lung carcinoma
ECACC Cellosaurus
Mono-Mac-6 Human White blood cells Myeloid metaplasic AML
DSMZ Cellosaurus
MRC-5 Medical Research Council cell strain 5 Human Lung (fetal) Fibroblast
ECACC Cellosaurus
MTD-1A Mouse Epithelium
Cellosaurus
MyEnd Myocardial Endothelial Mouse Endothelium
Cellosaurus
NCI-H69 Human Lung Lung carcinoma
ECACC Cellosaurus
NCI-H69/CPR Human Lung Lung carcinoma
ECACC Cellosaurus
NCI-H69/LX10 Human Lung Lung carcinoma
ECACC Cellosaurus
NCI-H69/LX20 Human Lung Lung carcinoma
ECACC Cellosaurus
NCI-H69/LX4 Human Lung Lung carcinoma
ECACC Cellosaurus
Neuro-2a Mouse Nerve/neuroblastoma
 Neuronal stem cells
ECACC Cellosaurus
NIH-3T3
NIH, 3-day transfer, inoculum 3 x 105 cells		 Mouse Embryo Fibroblast
ECACC Cellosaurus
NALM-1 Human Peripheral blood Blast-crisis CML
ATCC Cellosaurus
Neuro2a Mouse Nerve/neuroblastoma
 Neuronal stem cells
Cellosaurus
NK-92 Human Leukemia/lymphoma
ATCC Cellosaurus
NTERA-2 Human Lung metastasis Embryonal carcinoma
ECACC Cellosaurus
NW-145 Human Skin Melanoma
ESTDAB Cellosaurus
OK Opossum Kidney
Virginia opossum - Didelphis virginiana
 Kidney
ECACC Cellosaurus
OPCN / OPCT cell lines Human Prostate Range of prostate tumour lines
Cellosaurus
P3X63Ag8 Mouse Myeloma
ECACC Cellosaurus
PANC-1 Human Duct Epithelioid Carcinoma
ATCC Cellosaurus
PC12 Rat Adrenal medulla Pheochromocytoma
ECACC Cellosaurus
PC-3 Prostate Cancer-3 Human Bone metastasis Prostate carcinoma
ECACC Cellosaurus
Peer Human T cell leukemia
DSMZ Cellosaurus
PNT1A Human Prostate SV40-transformed tumour line
ECACC Cellosaurus
PNT2 Human Prostate SV40-transformed tumour line
ECACC Cellosaurus
Pt K2 The second cell line derived from Potorous tridactylis
Long-nosed potoroo - Potorous tridactylus
 Kidney Epithelial
ECACC Cellosaurus
Raji Human B lymphoma
 Lymphoblast-like
ECACC Cellosaurus
RBL-1 Rat Basophilic Leukemia-1 Rat Leukemia Basophil cell
ECACC Cellosaurus
RenCa Renal Carcinoma Mouse Kidney Renal carcinoma
ATCC Cellosaurus
RIN-5F Mouse Pancreas
ECACC Cellosaurus
RMA-S Mouse T cell tumour
Cellosaurus
S2 Schneider 2 Insect - Drosophila melanogaster
 Late stage (20–24 hours old) embryos
ATCC Cellosaurus
SaOS-2 Sarcoma OSteogenic-2 Human Bone Osteosarcoma
ECACC Cellosaurus
Sf21
Spodoptera frugiperda 21		 Insect (moth) - Spodoptera frugiperda
 Ovary
ECACC Cellosaurus
Sf9
Spodoptera frugiperda 9		 Insect (moth) - Spodoptera frugiperda
 Ovary
ECACC Cellosaurus
SH-SY5Y Human Bone marrow metastasis Neuroblastoma
ECACC Cellosaurus
SiHa Human Cervix epithelium Cervical carcinoma
ATCC Cellosaurus
SK-BR-3
Sloan-Kettering Breast cancer 3		 Human Breast Breast carcinoma
DSMZ Cellosaurus
SK-OV-3
Sloan-Kettering Ovarian cancer 3		 Human Ovary Ovarian carcinoma
ECACC Cellosaurus
SK-N-SH Human Brain Epithelial
ATCC Cellosaurus
T2 Human T cell leukemia/B cell line hybridoma
ATCC Cellosaurus
T-47D Human Breast Breast ductal carcinoma
ECACC Cellosaurus
T84 Human Lung metastasis Colorectal carcinoma
ECACC Cellosaurus
T98G Human Glioblastoma-astrocytoma Epithelium
ECACC Cellosaurus
THP-1 Human Monocyte Acute monocytic leukemia
ECACC Cellosaurus
U2OS Human Osteosarcoma Epithelial
ECACC Cellosaurus
U373 Human Glioblastoma-astrocytoma Epithelium
ECACC Cellosaurus
U87 Human Glioblastoma-astrocytoma Epithelial-like
ECACC Cellosaurus
U937 Human Leukemic monocytic lymphoma
ECACC Cellosaurus
VCaP Vertebral Cancer of the Prostate Human Vertebra metastasis Prostate carcinoma
ECACC Cellosaurus
Vero
Vero (truth)		 African green monkey - Chlorocebus sabaeus
 Kidney epithelium
ECACC Cellosaurus
VG-1 Human Primary effusion lymphoma
Cellosaurus
WM39 Human Skin Melanoma
ESTDAB Cellosaurus
WT-49 Human Lymphoblastoid
ECACC Cellosaurus
YAC-1 Mouse Lymphoma
ECACC Cellosaurus
YAR Human Lymphoblastoid EBV-transformed B cell
Human Immunology[60]ECACC Cellosaurus


See also



  • Biological immortality

  • Cell culture assays

  • Electric cell-substrate impedance sensing

  • List of contaminated cell lines

  • List of NCI-60 Cell Lines

  • List of breast cancer cell lines



References and notes





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Further reading


.mw-parser-output .refbegin{font-size:90%;margin-bottom:0.5em}.mw-parser-output .refbegin-hanging-indents>ul{list-style-type:none;margin-left:0}.mw-parser-output .refbegin-hanging-indents>ul>li,.mw-parser-output .refbegin-hanging-indents>dl>dd{margin-left:0;padding-left:3.2em;text-indent:-3.2em;list-style:none}.mw-parser-output .refbegin-100{font-size:100%}



  • Pacey L, Stead S, Gleave J, Tomczyk K, Doering L (2006). "Neural Stem Cell Culture: Neurosphere generation, microscopical analysis and cryopreservation". Protocol Exchange. doi:10.1038/nprot.2006.215.


  • Gilabert JA, Montalvo GB, Artalejo AR (2006). "Rat Chromaffin cells primary cultures: Standardization and quality assessment for single-cell assays". Protocol Exchange. doi:10.1038/nprot.2006.294.


  • Losardo RJ, Cruz-Gutiérrez R, Prates JC, Moscovici M, Rodríguez-Torres A, Arteaga-Martinez M (2015). "Sergey Fedoroff: A Pioneer of the Neuronal Regeneration. Tribute from the Pan American Association of Anatomy". International Journal of Morphology. 33 (2): 794. doi:10.4067/S0717-95022015000200059.


  • MacLeod RA, Dirks WG, Matsuo Y, Kaufmann M, Milch H, Drexler HG (November 1999). "Widespread intraspecies cross-contamination of human tumor cell lines arising at source". International Journal of Cancer. 83 (4): 555–63. doi:10.1002/(SICI)1097-0215(19991112)83:4<555::AID-IJC19>3.0.CO;2-2. PMID 10508494.


  • Masters JR (April 2002). "HeLa cells 50 years on: the good, the bad and the ugly". Nature Reviews. Cancer. 2 (4): 315–9. doi:10.1038/nrc775. PMID 12001993.


  • Witkowski JA (July 1983). "Experimental pathology and the origins of tissue culture: Leo Loeb's contribution". Medical History. 27 (3): 269–88. doi:10.1017/S0025727300042964. PMC 1139336. PMID 6353093.




External links








Library resources about
Cell culture



  • Resources in your library

  • Resources in other libraries


  • Table of common cell lines from Alberts 4th ed.

  • Cancer Cells in Culture

  • Evolution of Cell Culture Surfaces

  • Hypertext version of the Cell Line Data Base

  • Microcarrier Cell Culture Handbook by GE Healthcare Life Sciences


  • Cell Culture Applications - Resources including application notes and protocols to create an ideal environment for growing cells, right from the start.


  • Cell Culture Basics - Introduction to cell culture, covering topics such as laboratory set-up, safety and aseptic technique including basic cell culture protocols and video training

  • Database of Who's Who in Cell Culture and Related Research

  • Coriell Cell Repositories

  • Strategies for Protein Purification Handbook


  • An Introduction To Cell Culture. This webinar introduces the history, theory, basic techniques, and potential pit-falls of mammalian cell culture.


  • The National Centre for Cell Science (NCCS), Pune, India; national repository for cell lines/hybridomas etc.


  • Public Health England, Public Health England Culture Collections (ECACC)











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