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[10] Monolayer culture techniques

1979; Academic Press; Linguagem: Inglês

10.1016/s0076-6879(79)58131-2

1557-7988

James A. McAteer, W.H. Douglas,

Tissue Engineering and Regenerative Medicine

This chapter discusses the monolayer cell culture techniques, which are frequently established from single cell suspensions prepared by the enzymic dissociation of organ fragments. The cell preparation is inoculated into a culture vessel containing fluid medium and incubated in a controlled atmosphere. The resultant primary culture is a mixed cell population, which contains many of the cell types present in the tissue of origin. Cell separation and isolation methods such as density gradient centrifugation, electrophoresis, or affinity column separation can be applied to the initial cell suspension prior to culture. Fibroblasts replicate faster than most cell types. Dilution plating at clonal density five is used to separate cell types so that colonies which develop may be physically isolated and then subcultured as pure populations. Basic monolayer cell culture techniques are well established and accessible.