Types of cells

Ultimately all animal cells are derived from living tissue; however, some — such as HeLa cells — have been in culture for so long (about 50 years) that they have lost all resemblance to the

Mince tissue

Fig. 10.1 Generating a primary cell culture. Tissue is surgically removed from an anesthetized animal, and then minced and homogenized. Addition of collagenase breaks down extracellular collagen, but the enzyme does not attack intact cells. The cells are purified by filtration through a coarse mesh to remove large fragments, and then concentrated by deposition under a mild centrifugal field in a low-speed centrifuge. The pelleted cells are washed in various buffered media containing serum, and then can be subjected to differential low-speed centrifugation to partially separate cell types based on sedimentation rates (a function of cell size and density). Various fractions are plated onto culture dishes in the presence of a culture medium containing essential amino acids, vitamins, antibiotics, and serum. Cells grow as loose clumps that can be dispersed with mild trypsin treatment, and individual cell types then can be cultured.

Mince tissue

Culture enriched for slowly sedimenting single cells

Digest adhesions with collagenase

Digest adhesions with collagenase

Culture enriched for slowly sedimenting single cells

Filter and pellet

Filter and pellet

Culture enriched for dense cell clumps

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