用胚胎干细胞诱导的造血干/祖细胞重建小鼠造血功能研究

Reconstitute of the Hematopoietic Function of Mice Using Hematopoietic Stem/Progenitor Cells Derived from Embryonic Stem Cells

目的胚胎干细胞体外诱导分化出CD34+造血干/祖细胞,然后将其注射入致死量照射小鼠体内,以研究其重建造血功能。方法胚胎干细胞自由分化形成胚胎体后,加入造血刺激因子混合液以诱导产生CD34+造血干/祖细胞,将这些造血干/祖细胞注射入经致死剂量照射的小鼠,观察小鼠存活率的改变,并取存活2个月的小鼠骨髓和脾脏,PCR检测嵌合体形成。结果体外分化第13日CD34+造血干/祖细胞比例可高达17.36％,将这些细胞注射入致死量照射小鼠后,可提高存活率达86.67％,存活小鼠的骨髓和脾脏均检测到供体来源的细胞。结论胚胎干细胞体外分化获得CD34+造血干/祖细胞,且后者具有其相应的正常生物学功能。

Objective To induce Embryonic stem cells to differentiate into CD34+ hematopoietic stem/progenitor cells in vitro. To reconstitute the hematopoietic function of lethally irradiated mice. Methods Embryonic stem cells, formed embryoid bodies, were induced to differentiate into CD34+ hematopoietic stem/progenitor cells when cultured with hematopoietic growth factors. These CD34+ hematopoietic stem/progenitor cells were injected into lethally irradiated mice. PCR was performed to detect the sex-determining region of the Y-chromsome derived from the male donor cells in the bone marrow and the spleen cells of the transplanted female mice. Results The percentage of CD34+ cells could reach 17.36% on the thirteenth day. The survival rate of the group transplanted with CD34+ hematopoietic stem/progenitor cells was 86.67% . The donor cells were both found in the bone marrow and the spleen cells of the survived transplanted mice. Conclusion Embryonic stem cell could be induced to differentiate into CD34+ hematopoietic stem/progenitor cells in vitro, and these cells could reconstitute the hematopoietic function of the lethally irradiated mice.