ECACC Handbook

Fundamental Techniques in Cell Culture . . . a Laboratory Handbook

1.0   The ECACC and its Collections 2.0   Design and Equipment for the Cell Culture Laboratory 3.0   Safety Aspects of Cell Culture 4.0   Sourcing of Cell Lines 5.0   Main Types of Cell Culture     5.1   Primary Cultures     5.2   Continuous Cultures     5.3   Culture Morphology 6.0   The Cell Environment 7.0   Cryopreservation and Storage of Cell Lines 8.0   Good Cell Banking Practices 9.0   Quality Control Considerations 10.0   Authentication of Cell Lines 11.0   Alternative Cell Culture Systems 12.0   Cell Culture Protocols Glossary of Terms Who to Contact

5.0   Main Types of Cell Culture

5.1   Primary Cultures

Primary cultures are derived directly from excised, normal animal tissue and cultured either as an explant culture or following dissociation into a single cell suspension by enzyme digestion. Such cultures are initially heterogeneous but later become dominated by fibroblasts. The preparation of primary cultures is labor intensive and they can be maintained in vitro only for a limited period of time. During their relatively limited life span primary cells usually retain many of the differentiated characteristics of the cell in vivo.

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5.2   Continuous Cultures

Continuous cultures are comprised of a single cell type that can be serially propagated in culture either for a limited number of cell divisions (approximately thirty) or otherwise indefinitely. Cell lines of a finite life are usually diploid and maintain some degree of differentiation. The fact that such cell lines senesce after approximately thirty cycles of division means it is essential to establish a system of Master and Working banks in order to maintain such lines for long periods.

Continuous cell lines that can be propagated indefinitely generally have this ability because they have been transformed into tumor cells. Tumor cell lines are often derived from actual clinical tumors, but transformation may also be induced using viral oncogenes or by chemical treatments. Transformed cell lines present the advantage of almost limitless availability, but the disadvantage of having retained very little of the original in vivo characteristics.

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5.3   Culture Morphology

Morphologically cell cultures take one of two forms, growing either in suspension (as single cells or small free-floating clumps) or as a monolayer that is attached to the tissue culture flask. The form taken by a cell line reflects the tissue from which it was derived e.g. cell lines derived from blood (leukaemia, lymphoma) tend to grow in suspension whereas cells derived from solid tissue (lungs, kidney) tend to grow as monolayers. Attached cell lines can be classified as endothelial such as BAE-1 (Prod. No. 88031149-1v1), epithelial such as HeLa (Prod. No. 93021013-1v1), neuronal such as SH-SY5Y (Prod. No. 94030304-1v1) or fibroblasts such as MRC-5 (Prod. No. 84101801-1v1) and their morphology reflect the area within the tissue of origin.

Click here for Figure 2. Examples of Cell Morphology

The cell lines most commonly ordered from ECACC are listed in the table below (Table 1). The lines are classified by cell type.

Table 1. Commonly used cell lines of each culture type

There are some instances when cell cultures may grow as semi-adherent cells e.g. B95-8 where there appears to be a mixed population of attached and suspension cells. For these cell lines it is essential that both cell types are subcultured to maintain the heterogeneous nature of the culture.

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