Primary Cell Cultures and Immortal Cell Lines


Primary cell cultures can be established from tissue explants or, more usually, from disaggregated tissue samples. Cells that can be subcultured many times are called cell lines and examples have been isolated from diverse tissues and cell types. Continuous cell lines of unlimited lifespan can be generated by means of a number of immortalization techniques, although in most cases such cell lines have arisen spontaneously.

Keywords: primary cells; cell lines; immortalization; in vitro cell culture; standardization

Figure 1.

Typical appearance of the graph of additional cumulative population doublings of the finite cell line MRC‐5 from a starting culture at population doubling number 40 compared with a continuous cell line (e.g. HeLa). S indicates the onset of senescence.

Figure 2.

A typical master cell bank and working cell bank system (circles represent ampoules of preserved cells). Preparation of cell banks in this way ensures that all cultures provided from the working banks are identical in terms of subculture history (i.e. ‘age’). This type of system used in bona fide culture collections is a key component in promoting standardization in biological research and biotechnology. Reproduced with permission from Cryo Letters.



Andrews P (1998) Teratocarcinomas and human embryology: pluripotent human EC cell lines. APMIS 106: 158–167.

Coecke S, Balls M, Bowe G et al. (2005) Guidance on Good Cell Culture Practice. Alternatives to Laboratory Animals 33: 261–287.

Department of Health, UK (2002) A Code of Practice for the Production of Human‐derived Therapeutic Products. London: Medical Devices Agency (now MHRA). (ISBN 1 84182 582 4).

Evans M (1996) Embryonic stem cells: the route to experimental mammalian genetics. International Journal of Developmental Biology 1: 55S.

Freshney IR (1994) Culture of Animal Cells: A Manual of Basic Technique., 3rd edn. New York: Wiley‐Liss.

Hartung T, Balls M, Bardouille C et al. (2002) Good Cell Culture Practice Task Force Report. ATLA 30: 407–414. (see also A Report of the Second ECVAM Task Force on Good Cell Culture Practice. Draft report available at

Heins N, Englund MCO, Sjoblom C et al. (2004) Derivation, characterisation and differentiation of human embryonic stem cells. Stem Cells 22: 367–376.

Labosky PA, Barlow DP and Hogan BL (1994) Embryonic germ cell lines and their derivation from mouse primordial germ cells. Ciba Foundation Symposia 182: 157–168.

Patzke S, Lindeskog M, Munthe E and Aasheim HC (2002) Characterization of a novel human endogenous retrovirus, HERV‐H/F, expressed in human leukemia cell lines. Virology 303: 164–173.

Pera MF, Reubinoff B and Trounson A (2000) Human embryonic stem cells. Journal of Cell Science 113: 5–10.

Stacey GN (2000) Cell lines used in the manufacture of biologicals. In: Spier R (editor in chief) Encyclopedia of Cell Technology. New York: Wiley Interscience.

Stacey G, Masters JRW, Hay RJ et al. (2000) Cell contamination leads to inaccurate data: we must take action now. Nature 403: 356.

Stacey GN and Stacey AR (1999) Routine quality control testing for cell cultures. In: Kinchington D and Schinazi RF (eds) Methods in Molecular Medicine: Antiviral Chemotherapy Protocols, chap. 4. Totowa NJ: Humana Press.

Swim HE and Parker RF (1957) Culture characteristics of human fibroblasts propagated serially. American Journal of Hygiene 66: 235–243.

UKCCCR (1999) UKCCCR Guidelines for the Use of Cell Lines in Cancer Research. London: UKCCCR.

WHO Expert Committee on Biological Standardization and Executive Board (1998) Requirements for the Use of Animal Cells as In Vitro Substrates for the Production of Biologicals (Requirements for Biological Substances No. 50). WHO Technical Report Series No. 878. Geneva: World Health Organization.

Further Reading

Schaeffer WI (1990) Nomenclature of cells in culture. In vitro 26: 39–63.

Stacey GN, Doyle A and Hambleton PH (1998) Safety in Cell and Tissue Culture. Dordrecht, The Netherlands: Kluwer Academic.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Stacey, Glyn(Jan 2006) Primary Cell Cultures and Immortal Cell Lines. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0003960]