Cell culture
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, growth factors, hormones, and gases, and regulates the physio-chemical environment. Most cells require a surface or an artificial substrate whereas others can be grown free floating in culture medium. 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. 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 separated from their original tissue source became more robust in the middle 20th century.
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. 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. 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.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
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. 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.Cells are grown and maintained at an appropriate temperature and gas mixture 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.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, 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. 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, 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, and using purified nutrient concentrates derived from serum in place of whole animal serum for cell culture.
Plating density 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.
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 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 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.
Components of cell culture media
Typical Growth conditions
Cell line cross-contamination
Cell line cross-contamination can be a problem for scientists working with cultured cells. Studies suggest anywhere from 15–20% of the time, cells used in experiments have been misidentified or contaminated with another cell line. 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. Major cell line repositories, including the American Type Culture Collection, the European Collection of Cell Cultures and the German Collection of Microorganisms and Cell Cultures, have received cell line submissions from researchers that were misidentified by them. 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. ATCC uses short tandem repeat DNA fingerprinting to authenticate its cell lines.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 typing, chromosomal analysis, karyotyping, morphology and STR analysis.
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 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.
Manipulation of cultured 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 cabinet or laminar flow cabinet to exclude contaminating micro-organisms. Antibiotics and antifungals 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
Passaging 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
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
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.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 of an immunised animal are combined with an immortal myeloma cell line to produce a hybridoma which has the antibody specificity of the primary lymphocyte and the immortality of the myeloma. Selective growth medium 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 and, after this, they lose their ability to proliferate.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. Stem cell culture is also used to harvest the molecules and exosomes that the stem cells release for the purposes of therapeutic development.Biological products produced by recombinant DNA technology in animal cell cultures include enzymes, synthetic hormones, immunobiologicals, and anticancer agents. Although many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins that are glycosylated 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, 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.
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 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, a concept that has led to discoveries in fields such as:
- Stem cell self-renewal
- Lineage specification
- Cancer cell phenotype
- Fibrosis
- Hepatocyte function
- Mechanosensing
Cell culture in three dimensions
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, transformed Human Carcinoma, and Mink Lung Epithelium 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.
3D cell culture in hydrogels
As the natural extracellular matrix 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. 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 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
s 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 as a vector,and novel adjuvants.
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 senescence occurs. Cultured primary cells have been extensively used in research, as is the case of fish keratocytes in cell migration studies.Plant cell culture methods
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 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
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
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
- H295R
- HeLa
- KBM-7
- LNCaP
- MCF-7
- MDA-MB-468
- PC3
- SaOS-2
- SH-SY5Y
- T-47D
- THP-1
- U87
- National Cancer Institute's 60 cancer cell line panel
- Vero
- MC3T3
- GH3
- PC12
- Tobacco BY-2 cells
- Dog MDCK kidney epithelial
- Xenopus A6 kidney epithelial
- Zebrafish AB9
List of cell lines
| Cell line | Meaning | Organism | Origin tissue | Morphology | Links |
| 3T3-L1 | "3-day transfer, inoculum 3 x 10^5 cells" | Mouse | Embryo | Fibroblast | |
| 4T1 | Mouse | Mammary gland | |||
| 9L | Rat | Brain | Glioblastoma | ||
| A172 | Human | Brain | Glioblastoma | ||
| A20 | Mouse | B lymphoma | B lymphocyte | ||
| A253 | Human | Submandibular duct | Head and neck carcinoma | ||
| A2780 | Human | Ovary | Ovarian carcinoma | ||
| A2780ADR | Human | Ovary | Adriamycin-resistant derivative of A2780 | ||
| A2780cis | Human | Ovary | Cisplatin-resistant derivative of A2780 | ||
| A431 | Human | Skin epithelium | Squamous cell carcinoma | ||
| A549 | Human | Lung | Lung carcinoma | ||
| AB9 | Zebrafish | Fin | Fibroblast | ||
| AHL-1 | Armenian Hamster Lung-1 | Hamster | Lung | ||
| ALC | Mouse | Bone marrow | Stroma | ||
| B16 | Mouse | Melanoma | |||
| B35 | Rat | Neuroblastoma | |||
| BCP-1 | Human | PBMC | HIV+ primary effusion lymphoma | ||
| BEAS-2B | Bronchial epithelium + Adenovirus 12-SV40 virus hybrid | Human | Lung | Epithelial | |
| bEnd.3 | Brain Endothelial 3 | Mouse | Brain/cerebral cortex | Endothelium | |
| BHK-21 | Baby Hamster Kidney-21 | Hamster | Kidney | Fibroblast | |
| BOSC23 | Packaging cell line derived from HEK 293 | Human | Kidney | Epithelium | |
| BT-20 | Breast Tumor-20 | Human | Breast epithelium | Breast carcinoma | |
| BxPC-3 | Biopsy xenograft of Pancreatic Carcinoma line 3 | Human | Pancreatic adenocarcinoma | Epithelial | |
| C2C12 | Mouse | Myoblast | |||
| C3H-10T1/2 | Mouse | Embryonic mesenchymal cell line | |||
| C6 | Rat | Brain astrocyte | Glioma | ||
| C6/36 | Insect - Asian tiger mosquito | Larval tissue | |||
| Caco-2 | Human | Colon | Colorectal carcinoma | ||
| Cal-27 | Human | Tongue | Squamous cell carcinoma | ||
| Calu-3 | Human | Lung | Adenocarcinoma | ||
| CGR8 | Mouse | Embryonic stem cells | |||
| CHO | Chinese Hamster Ovary | Hamster | Ovary | Epithelium | |
| CML T1 | Chronic myeloid leukemia T lymphocyte 1 | Human | CML acute phase | T cell leukemia | |
| CMT12 | Canine Mammary Tumor 12 | Dog | Mammary gland | Epithelium | |
| COR-L23 | Human | Lung | Lung carcinoma | ||
| COR-L23/5010 | Human | Lung | Lung carcinoma | ||
| COR-L23/CPR | Human | Lung | Lung carcinoma | ||
| COR-L23/R23- | Human | Lung | Lung carcinoma | ||
| COS-7 | Cercopithecus aethiops, origin-defective SV-40 | Old World monkey - Cercopithecus aethiops | Kidney | Fibroblast | |
| COV-434 | Human | Ovary | Ovarian granulosa cell carcinoma | ||
| CT26 | Mouse | Colon | Colorectal carcinoma | ||
| D17 | Dog | Lung metastasis | Osteosarcoma | ||
| DAOY | Human | Brain | Medulloblastoma | ||
| DH82 | Dog | Histiocytosis | Monocyte/macrophage | ||
| DU145 | Human | Androgen insensitive prostate carcinoma | |||
| DuCaP | Dura mater cancer of the Prostate | Human | Metastatic prostate carcinoma | Epithelial | |
| E14Tg2a | Mouse | Embryonic stem cells | |||
| EL4 | Mouse | T cell leukemia | |||
| EM-2 | Human | CML blast crisis | Ph+ CML line | ||
| EM-3 | Human | CML blast crisis | Ph+ CML line | ||
| EMT6/AR1 | Mouse | Mammary gland | Epithelial-like | ||
| EMT6/AR10.0 | Mouse | Mammary gland | Epithelial-like | ||
| FM3 | Human | Lymph node metastasis | Melanoma | ||
| GL261 | Glioma 261 | Mouse | Brain | Glioma | |
| H1299 | Human | Lung | Lung carcinoma | ||
| HaCaT | Human | Skin | Keratinocyte | ||
| HCA2 | Human | Colon | Adenocarcinoma | ||
| HEK 293 | Human Embryonic Kidney 293 | Human | Kidney | Epithelium | |
| HEK 293T | HEK 293 derivative | Human | Kidney | Epithelium | |
| HeLa | "Henrietta Lacks" | Human | Cervix epithelium | Cervical carcinoma | |
| Hepa1c1c7 | Clone 7 of clone 1 hepatoma line 1 | Mouse | Hepatoma | Epithelial | |
| Hep G2 | Human | Liver | Hepatoblastoma | ||
| High Five | Insect - Trichoplusia ni | Ovary | |||
| HL-60 | Human Leukemia-60 | Human | Blood | Myeloblast | |
| HT-1080 | Human | Fibrosarcoma | |||
| HT-29 | Human | Colon epithelium | Adenocarcinoma | ||
| J558L | Mouse | Myeloma | B lymphocyte cell | ||
| Jurkat | Human | White blood cells | T cell leukemia | ||
| JY | Human | Lymphoblastoid | EBV-transformed B cell | ||
| K562 | Human | Lymphoblastoid | CML blast crisis | ||
| KBM-7 | Human | Lymphoblastoid | CML blast crisis | ||
| KCL-22 | Human | Lymphoblastoid | CML | ||
| KG1 | Human | Lymphoblastoid | AML | ||
| Ku812 | Human | Lymphoblastoid | Erythroleukemia | ||
| KYO-1 | Kyoto-1 | Human | Lymphoblastoid | CML | |
| L1210 | Mouse | Lymphocytic leukemia | Ascitic fluid | ||
| L243 | Mouse | Hybridoma | Secretes L243 mAb | ||
| LNCaP | Lymph Node Cancer of the Prostate | Human | Prostatic adenocarcinoma | Epithelial | |
| MA-104 | Microbiological Associates-104 | African Green Monkey | Kidney | Epithelial | |
| MA2.1 | Mouse | Hybridoma | Secretes MA2.1 mAb | ||
| Ma-Mel 1, 2, 3....48 | Human | Skin | A range of melanoma cell lines | ||
| MC-38 | Mouse Colon-38 | Mouse | Colon | Adenocarcinoma | |
| MCF-7 | Michigan Cancer Foundation-7 | Human | Breast | Invasive breast ductal carcinoma ER+, PR+ | |
| MCF-10A | Michigan Cancer Foundation-10A | Human | Breast epithelium | ||
| MDA-MB-157 | M.D. Anderson - Metastatic Breast-157 | Human | Pleural effusion metastasis | Breast carcinoma | |
| MDA-MB-231 | M.D. Anderson - Metastatic Breast-231 | Human | Pleural effusion metastasis | Breast carcinoma | |
| MDA-MB-361 | M.D. Anderson - Metastatic Breast-361 | Human | Melanoma | ||
| MDA-MB-468 | M.D. Anderson - Metastatic Breast-468 | Human | Pleural effusion metastasis | Breast carcinoma | |
| MDCK II | Madin Darby Canine Kidney II | Dog | Kidney | Epithelium | |
| MG63 | Human | Bone | Osteosarcoma | ||
| MIA PaCa-2 | Human | Prostate | Pancreatic Carcinoma | ||
| MOR/0.2R | Human | Lung | Lung carcinoma | ||
| Mono-Mac-6 | Human | White blood cells | Myeloid metaplasic AML | ||
| MRC-5 | Medical Research Council cell strain 5 | Human | Lung | Fibroblast | |
| MTD-1A | Mouse | Epithelium | |||
| MyEnd | Myocardial Endothelial | Mouse | Endothelium | ||
| NCI-H69 | Human | Lung | Lung carcinoma | ||
| NCI-H69/CPR | Human | Lung | Lung carcinoma | ||
| NCI-H69/LX10 | Human | Lung | Lung carcinoma | ||
| NCI-H69/LX20 | Human | Lung | Lung carcinoma | ||
| NCI-H69/LX4 | Human | Lung | Lung carcinoma | ||
| Neuro-2a | Mouse | Nerve/neuroblastoma | Neuronal stem cells | ||
| NIH-3T3 | NIH, 3-day transfer, inoculum 3 x 105 cells | Mouse | Embryo | Fibroblast | |
| NALM-1 | Human | Peripheral blood | Blast-crisis CML | ||
| NK-92 | Human | Leukemia/lymphoma | |||
| NTERA-2 | Human | Lung metastasis | Embryonal carcinoma | ||
| NW-145 | Human | Skin | Melanoma | ||
| OK | Opossum Kidney | Virginia opossum - Didelphis virginiana | Kidney | ||
| OPCN / OPCT cell lines | Human | Prostate | Range of prostate tumour lines | ||
| P3X63Ag8 | Mouse | Myeloma | |||
| PANC-1 | Human | Duct | Epithelioid Carcinoma | ||
| PC12 | Rat | Adrenal medulla | Pheochromocytoma | ||
| PC-3 | Prostate Cancer-3 | Human | Bone metastasis | Prostate carcinoma | |
| Peer | Human | T cell leukemia | |||
| PNT1A | Human | Prostate | SV40-transformed tumour line | ||
| PNT2 | Human | Prostate | SV40-transformed tumour line | ||
| Pt K2 | The second cell line derived from Potorous tridactylis | Long-nosed potoroo - Potorous tridactylus | Kidney | Epithelial | |
| Raji | Human | B lymphoma | Lymphoblast-like | ||
| RBL-1 | Rat Basophilic Leukemia-1 | Rat | Leukemia | Basophil cell | |
| RenCa | Renal Carcinoma | Mouse | Kidney | Renal carcinoma | |
| RIN-5F | Mouse | Pancreas | |||
| RMA-S | Mouse | T cell tumour | |||
| S2 | Schneider 2 | Insect - Drosophila melanogaster | Late stage embryos | ||
| SaOS-2 | Sarcoma OSteogenic-2 | Human | Bone | Osteosarcoma | |
| Sf21 | Spodoptera frugiperda 21 | Insect - Spodoptera frugiperda | Ovary | ||
| Sf9 | Spodoptera frugiperda 9 | Insect - Spodoptera frugiperda | Ovary | ||
| SH-SY5Y | Human | Bone marrow metastasis | Neuroblastoma | ||
| SiHa | Human | Cervix epithelium | Cervical carcinoma | ||
| SK-BR-3 | Sloan-Kettering Breast cancer 3 | Human | Breast | Breast carcinoma | |
| SK-OV-3 | Sloan-Kettering Ovarian cancer 3 | Human | Ovary | Ovarian carcinoma | |
| SK-N-SH | Human | Brain | Epithelial | ||
| T2 | Human | T cell leukemia/B cell line hybridoma | |||
| T-47D | Human | Breast | Breast ductal carcinoma | ||
| T84 | Human | Lung metastasis | Colorectal carcinoma | ||
| T98G | Human | Glioblastoma-astrocytoma | Epithelium | ||
| THP-1 | Human | Monocyte | Acute monocytic leukemia | ||
| U2OS | Human | Osteosarcoma | Epithelial | ||
| U373 | Human | Glioblastoma-astrocytoma | Epithelium | ||
| U87 | Human | Glioblastoma-astrocytoma | Epithelial-like | ||
| U937 | Human | Leukemic monocytic lymphoma | |||
| VCaP | Vertebral Cancer of the Prostate | Human | Vertebra metastasis | Prostate carcinoma | |
| Vero | From Esperanto: verda reno | African green monkey - Chlorocebus sabaeus | Kidney epithelium | ||
| VG-1 | Human | Primary effusion lymphoma | |||
| WM39 | Human | Skin | Melanoma | ||
| WT-49 | Human | Lymphoblastoid | |||
| YAC-1 | Mouse | Lymphoma | |||
| YAR | Human | Lymphoblastoid | EBV-transformed B cell |