Schaap G H, Verkerk A, Vijg J, Jongkind J F
Cytometry. 1983 May;3(6):408-13. doi: 10.1002/cyto.990030604.
Undifferentiated mouse teratocarcinoma cells were cocultivated with differentiated mouse endoderm cells in order to study the possible induction of teratocarcinoma cell differentiation. A difference in DNA content between the two cell types was experimentally introduced to enable the reisolation of the teratocarcinoma cells after cocultivation. Pseudotetraploid (2s) endoderm cell lines were produced from pseudodiploid (1s) cells by treatment of these cells with cytochalasin B and flow sorting of tetraploid cells, using Hoechst 33342 as a viable DNA stain, with subsequent cloning of sorted single cells. In model experiments, where mixtures of 1s teratocarcinoma and 2s endoderm cells were stained with Hoechst 33342, the teratocarcinoma cells could be reisolated with a purity of about 97%. After a cocultivation period of 24 days viable teratocarcinoma cells could be isolated from the cocultivation mixture with a purity of 95%. Two dimensional analysis of the protein pattern of these cells indicated that cocultivation did not induce a differentiated (endoderm) pattern. Therefore according to this analysis the teratocarcinoma cells were not induced to differentiate during a 24 day cocultivation period. The method described offers excellent possibilities for studying cell-cell interaction in vitro.
为了研究畸胎癌细胞分化的可能诱导情况，将未分化的小鼠畸胎癌细胞与已分化的小鼠内胚层细胞进行共培养。通过实验引入两种细胞类型之间的DNA含量差异，以便在共培养后重新分离出畸胎癌细胞。通过用细胞松弛素B处理假二倍体（1s）细胞并对四倍体细胞进行流式分选，使用Hoechst 33342作为活细胞DNA染色剂，随后对分选的单细胞进行克隆，从假二倍体细胞产生假四倍体（2s）内胚层细胞系。在模型实验中，用Hoechst 33342对1s畸胎癌和2s内胚层细胞的混合物进行染色，畸胎癌细胞可以以约97%的纯度重新分离出来。共培养24天后，可从共培养混合物中分离出活力良好的畸胎癌细胞，纯度为95%。对这些细胞蛋白质模式的二维分析表明，共培养并未诱导出分化（内胚层）模式。因此，根据该分析，在24天的共培养期内，畸胎癌细胞未被诱导分化。所描述的方法为体外研究细胞间相互作用提供了极好的可能性。
