Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer.

摘要: Two methods for simple and rapid plating of single HeLa cells, human, carcinomatous are described. These result in growth formation colonies from each cell. One these procedures uses irradiated, non-multiplying "feeder" cells to condition the medium. The second requires more gentle handling but otherwise is virtually same as that used bacteria on semisolid, nutrient media. By extension methods, it possible isolate mutant grow pure clonal stocks animal cells. genetically uniform strains much homogeneous their behavior than parental cell population. Growth curves obtained developing highly reproducible. most active so far isolated display a generation time 18 20 hours. In pooled human serum assume stretched, ameboid form, with marked motility; whereas variety non-human sera results tightly packed, columnar, epithelial-like morphology. two types possess volumes, nuclear cross-sections, efficiencies, times which identical within experimental error, widely different cross-sectional areas, suggesting basic change occurs surface. It conceivable this may be related enables compact tumor become invasive. Animal subjected standard trypsinization involve mechanical trauma repeated washings incomplete media leak large amounts P suffer impaired ability reproduce Application described paper tool quantitative study normal mammalian growth, physiology, genetics, biochemistry, response drugs, viruses, high energy radiation, other agents have been indicated.