Culture conditions induce the appearance of immortalized C3H mouse cell lines.
Mouse fibroblasts were cultured by three different procedures: (a) changing the 0.2 ml/cm2 of growth medium every 2nd d and seeding 1 X 10(5) cells/cm2 after confluency; (b) changing the 0.4 ml/cm2 of growth medium only at subculture performed at confluency by a 1:2 split and keeping the bottles incubated on a rocking platform; (c) the same as Method b but keeping the bottles stationary throughout culture. By Method a no lines were immortalized over 36 experiments whereas Method b gave 1/4 immortalized lines and Method c gave 10:12 immortalized lines. Cells always went into crisis at the 9th to 11th doubling. Immortalized lines had a tetraploid DNA content.
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D003018:Clot Retraction; D003470:Culture Media; D005347:Fibroblasts / Q000166:cytology; D051379:Mice; D008809:Mice, Inbred C3H
D003470:Culture Media