小鼠孤雌胚胎干细胞系的建立及生物学特性鉴定

Derivation and Characterization of Mouse Parthenogenetic Embryonic Stem Cell Lines

目的:探讨建立合适的小鼠孤雌胚胎干细胞建系方法。方法:采用氯化锶联合细胞松弛素B激活B6D2F1杂交小鼠卵母细胞,所获得的囊胚与桑椹胚分别用于孤雌胚胎干细胞的建系,观察两者的建系成功率。结果:共建立了12株小鼠孤雌胚胎干细胞系,这些细胞SSEA-1抗原阳性,SSEA-4,TRA-1-81,TRA-1-60表面抗原阴性,具有AKP活性,保持正常染色体核型,体内外分化分别形成畸胎瘤和拟胚体。结论:采用囊胚和去透明带的桑葚胚建立孤雌胚胎干细胞系获得成功。该方法为人类纯合子的胚胎干细胞建系提供基础,在自体细胞治疗领域中具有潜在的应用价值。

Objective： To establish the optimum method of deriving mouse parthenogenetic embryonic stem cells, Methods： The B6D2F1 hybrid mouse oocytes were parthenogenetic activated with strontium chloride （SrCI2） and cytochalasin B（CB）. The parthenogenetic blastocysts and morulas were seeded on MEF feeder respectively to examine the primary ICM appearance and compare the derivation rate, Results： Twelve batches of mouse parthenogenetic embryonic stem cells were established. The established cells were positive for SSEA-1 and negative for SSEA-4, TRA-1-81, TRA-1-60 markers, They showed a high level of AKP activity, kept the normal Karyotypes,formed both embryoid bodies in vitro and teratomas in vivo, Conclusions： Mouse parthenogenetic embryonic stem cell lines were successfially derived from blastocysts and morulas without zona pellucida. This technology should provide a plateform for deriving human homozygous stem cells that will have a potential value in autologous cell therapy,