Kyuwa S, Xiao Y, Toyoda Y, Sato E
Department of Reproductive and Developmental Biology, University of Tokyo, Japan.
Exp Anim. 1997 Jan;46(1):11-6. doi: 10.1538/expanim.46.11.
Embryoid bodies (EB) were formed by TT2 embryonic stem (ES) cells in vitro. ES-like cell lines (ESLC) were established by culturing cells obtained by disaggregation of EB at 4, 8 and 20 days after culture, and designated ESLC4, ESLC8 and ESLC20, respectively. Flow cytometric analysis indicated that the cell surface expression of Le(a) on ESLC was less than that of original TT2 ES cells, but the expression of L-CAM was comparable. After suspension culture, all of the ESLC cells formed cystic EB in vitro. In addition, some ESLC4- and ESLC8-derived EB showed signs of beating. Although coat color chimeras were able to be produced with ESCL4 at a lower rate than parental ES cells, the cells did not contribute to germ line cells in chimeras. These results suggest that the ESLC had less pluripotent than parental ES cells and also that EB formation is not useful in obtaining pluripotent cells.
类胚体（EB）由TT2胚胎干细胞（ES）在体外形成。通过培养培养4、8和20天后从EB解离获得的细胞建立了类ES细胞系（ESLC），分别命名为ESLC4、ESLC8和ESLC20。流式细胞术分析表明，ESLC上Le(a)的细胞表面表达低于原始TT2 ES细胞，但L-CAM的表达相当。悬浮培养后，所有ESLC细胞在体外形成囊性EB。此外，一些源自ESLC4和ESLC8的EB显示出跳动迹象。虽然与亲代ES细胞相比，用ESCL4产生毛色嵌合体的效率较低，但这些细胞在嵌合体中并未对生殖系细胞有贡献。这些结果表明，ESLC的多能性低于亲代ES细胞，并且EB形成对于获得多能细胞并无用处。
