Characterization of cells and their applications in cell culture
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Primary cultures are derived directly from excised, normal animal tissue and cultures 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 fi broblasts. The preparation of primary cultures is labour intensive and they can be maintained in vitro only for a limited period of time. During their relatively limited lifespan primary cells usually retain many of the differentiated characteristics of the cell in vivo.
Important Note: Primary cultures by defi nition have not been passaged, as soon as they are passaged they become a cell line and are no longer primary. ‘Primary’ cells sourced from most suppliers are in fact low-passage cell lines.
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 indefi nitely. Cell lines of a fi nite 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 indefi nitely generally have this ability because they have been transformed into tumour cells. Tumour cell lines are often derived from actual clinical tumours, 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.
Culture Morphology
In terms of growth mode cell cultures take one of two forms, growing either in suspension (as single cells or small free fl oating clumps) or as a monolayer that is attached to the tissue culture fl ask. The form taken by a cell line refl ects 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 classifi ed as endothelial such as BAE-1, epithelial such as HeLa, neuronal such as SH-SY5Y, or fi broblasts such as MRC-5 and their morphology refl ects the area within the tissue of origin..
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Primary cultures are derived directly from excised, normal animal tissue and cultures 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 fi broblasts. The preparation of primary cultures is labour intensive and they can be maintained in vitro only for a limited period of time. During their relatively limited lifespan primary cells usually retain many of the differentiated characteristics of the cell in vivo.
Important Note: Primary cultures by defi nition have not been passaged, as soon as they are passaged they become a cell line and are no longer primary. ‘Primary’ cells sourced from most suppliers are in fact low-passage cell lines.
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 indefi nitely. Cell lines of a fi nite 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 indefi nitely generally have this ability because they have been transformed into tumour cells. Tumour cell lines are often derived from actual clinical tumours, 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.
Culture Morphology
In terms of growth mode cell cultures take one of two forms, growing either in suspension (as single cells or small free fl oating clumps) or as a monolayer that is attached to the tissue culture fl ask. The form taken by a cell line refl ects 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 classifi ed as endothelial such as BAE-1, epithelial such as HeLa, neuronal such as SH-SY5Y, or fi broblasts such as MRC-5 and their morphology refl ects the area within the tissue of origin..
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