Kubbies M, Friedl R, Bettecken T, Hoehn H
Cytometry. 1986 Nov;7(6):551-7. doi: 10.1002/cyto.990070609.
Primary cultures derived from lung and renal tissue of the newborn harvest mouse (Micromys minutus) were serially passaged in media supplemented with epidermal growth factor, hydrocortisone, transferrin, insulin, and triiodothyronine. Although these growth factor supplements eliminated the growth crisis commonly encountered during the initial stages of murine primary cultures, the original diploid cell fraction clearly underwent such a "crisis"; the truly diploid cells invariably disappeared as these cultures reached 20 to 40 population doublings. They were replaced, either gradually or precipitously, by various heteroploid cell fractions. In three of four independent cultures, these "established" cells were hypotetraploid and appeared to be derived from a small number of progenitors already present during the very early (precrisis) culture stages. In contrast to rather frequent DNA changes displayed by clones and subclones derived from the various heteroploid cell lineages, the predominant components of the established mass cultures displayed a highly constant DNA fluorescence pattern. Our results suggest that primary murine cell cultures develop heteroploid cell lineages even if the initial growth crisis is mitigated by growth factor supplements. These heteroploid cells appear to respond more efficiently to stimulation by various growth factors than the primary diploid cell population.
从新生收获小鼠（小家鼠）的肺和肾组织中获得的原代培养物，在补充了表皮生长因子、氢化可的松、转铁蛋白、胰岛素和三碘甲状腺原氨酸的培养基中进行连续传代培养。尽管这些生长因子补充剂消除了小鼠原代培养初始阶段常见的生长危机，但原始的二倍体细胞部分显然经历了这样的“危机”；随着这些培养物达到20至40次群体倍增，真正的二倍体细胞总是消失。它们逐渐或突然被各种异倍体细胞部分所取代。在四个独立培养物中的三个中，这些“建立的”细胞是亚四倍体，似乎源自极早期（危机前）培养阶段就已存在的少量祖细胞。与源自各种异倍体细胞谱系的克隆和亚克隆所显示的相当频繁的DNA变化形成对比的是，建立的大量培养物的主要成分显示出高度恒定的DNA荧光模式。我们的结果表明，即使初始生长危机通过生长因子补充剂得到缓解，原代小鼠细胞培养物仍会发展出异倍体细胞谱系。这些异倍体细胞似乎比原代二倍体细胞群体对各种生长因子的刺激反应更有效。
