Coulombel L, Huyhn A, Izac B
Unité INSERM 362, Institut Gustave-Roussy, Villejuif.
C R Seances Soc Biol Fil. 1995;189(4):611-5.
In a search for assays that might facilitate identification of pluripotent stem cells with extended potentialities, we analysed the properties of hematopoietic progenitor cells detected in the extraembryonic yolk sac and in the intraembryonic part of human embryos between approximately 28 and 45 days of development. Cells from the yolk sac, the liver rudiment and the remainder of the embryo were plated in semi solid methylcellulose colony-assays supplemented with combinations of cytokines. Large BFU-E-derived colonies as well as granulocytic colonies were detected in every yolk sac sample. Interestingly, progenitor cells were also detected in the intraembryonic part, outside the liver and a subclass of these progenitors were detected that generated large granulomacrophagic colonies capable of generating secondary colonies when replated. These were preferentially located in the embryo. Colony-assays initiated with CD34+ cells sorted from the different tissues confirmed these data. These results first indicate that embryonic progenitors exhibit unique phenotypic features, and second, analysis of the distribution of progenitors between the different tissues may suggest the existence of other sites of hematopoietic production. More detailed analysis of the potentialities of these progenitors should now be assessed in vitro in cocultures assays and in vivo by reconstituting immunodeficient mice.
为了寻找可能有助于鉴定具有扩展潜能的多能干细胞的检测方法,我们分析了在人类胚胎发育约28至45天期间,在胚外卵黄囊和胚内部分检测到的造血祖细胞的特性。将来自卵黄囊、肝原基和胚胎其余部分的细胞接种到添加了细胞因子组合的半固体甲基纤维素集落检测中。在每个卵黄囊样本中都检测到了大量源自爆式红系集落形成单位的集落以及粒细胞集落。有趣的是,在胚内部分、肝脏以外也检测到了祖细胞,并且检测到了这些祖细胞的一个亚类,它们能形成大型粒巨噬细胞集落,重新接种时能够产生次级集落。这些集落优先位于胚胎中。用从不同组织中分选的CD34 +细胞启动的集落检测证实了这些数据。这些结果首先表明胚胎祖细胞表现出独特的表型特征,其次,对不同组织之间祖细胞分布的分析可能提示存在其他造血产生部位。现在应该通过共培养检测在体外以及通过重建免疫缺陷小鼠在体内对这些祖细胞的潜能进行更详细的分析。