Eistetter Harald R
Friedrich Miescher Laboratorium der Max Planck Gesellschaft, Spemannstrasse 37/39, D-7400 Tübingen, FRG.
Dev Growth Differ. 1989 Jun;31(3):275-282. doi: 10.1111/j.1440-169X.1989.00275.x.
Mouse pluripotent embryonal stem (ES) cell lines hitherto have been conventionally isolated from the 'inner cell mass' of mouse blastocysts. In this report, I describe a new and simplified method for establishing pluripotent cell lines from mouse morulae of the 16- to 20-cell stage, which were disaggregated by the use of EDTA. From 17 cell lines established in such a way, 7 were characterized with respect to their differentiation potential: (i) When injected into syngeneic mice, the cells gave rise to solid, fully differentiated teratomas representing derivatives of all three germ layers. (ii) When cultured in suspension in vitro, the cells were able to differentiate into complex organized 'embryoid bodies' analogous to mouse early postimplantation embryos. These results strongly imply that embryonal stem cell lines isolated from mouse morulae are highly homologous to conventionally isolated ES cells. In addition, my results indicate that murine pluripotent embryonal stem (ES) cell lines can be derived with more ease and higher efficiency from disaggregated morulae than from the 'inner cell mass' of blastocysts.
迄今为止，小鼠多能胚胎干细胞系通常是从囊胚的“内细胞团”中分离得到的。在本报告中，我描述了一种新的、简化的方法，用于从16至20细胞阶段的小鼠桑椹胚建立多能细胞系，这些桑椹胚通过使用乙二胺四乙酸（EDTA）进行解离。在以这种方式建立的17个细胞系中，有7个细胞系的分化潜能得到了表征：（i）当将这些细胞注射到同基因小鼠体内时，它们会形成实体的、完全分化的畸胎瘤，代表所有三个胚层的衍生物。（ii）当在体外悬浮培养时，这些细胞能够分化成类似于小鼠植入后早期胚胎的复杂有组织的“胚状体”。这些结果强烈表明，从小鼠桑椹胚分离得到的胚胎干细胞系与传统分离的胚胎干细胞高度同源。此外，我的结果表明，与从囊胚的“内细胞团”中获得相比，从小鼠桑椹胚解离物中更容易且更高效地获得小鼠多能胚胎干细胞系。
