Buc-Caron M H, Launay J M, Marie P J, Kellermann O
Institut Pasteur, Paris, France.
Reprod Nutr Dev. 1990;30(3):309-16.
With the aim of immortalizing embryonic cells fixed at early embryonic stages, various plasmids carrying the SV40 early region were introduced into the mouse embryonal carcinomas (EC) F9 and 1003. Only the construction PK4, in which the SV40 oncogenes are placed under the control of the adenovirus E1A promoter, led to the immortalization of the cells at the onset of differentiation. Clones corresponding to committed precursors of each embryonic lineage (neuroectoderm, mesoderm and endoderm) were then selected with high efficiency according to the following strategy: selection of immature cells which: have lost EC cell markers, keep a stable phenotype, are immortalized by the expression of the SV40 oncogenes and are still able to differentiate along a restricted lineage in vitro or in vivo. Examples of an endodermal precursor (H7) which differentiates into extraembryonic and embryonic endoderm, of a neuroectodermic clone (ICII) committed to a serotoninergic differentiation, and of a mesodermal osteogenic clone (CI) which gives rise to bone in vivo and in vitro, are given.
为了使固定在胚胎早期阶段的胚胎细胞永生化,将携带SV40早期区域的各种质粒导入小鼠胚胎癌(EC)F9和1003细胞系。只有构建体PK4(其中SV40癌基因置于腺病毒E1A启动子的控制之下)能在细胞开始分化时使其永生化。然后根据以下策略高效筛选出对应于每个胚胎谱系(神经外胚层、中胚层和内胚层)定向前体细胞的克隆:选择具有以下特征的未成熟细胞:已丧失EC细胞标志物、保持稳定表型、通过SV40癌基因的表达而永生化且仍能够在体外或体内沿着特定谱系分化。文中给出了如下示例:一个内胚层前体细胞(H7)可分化为胚外和胚胎内胚层;一个神经外胚层克隆(ICII)定向分化为5-羟色胺能细胞;一个中胚层成骨克隆(CI)在体内和体外均可形成骨组织。