小鼠孤雌生殖胚胎干细胞的分离及分化潜能研究

用免疫外科法从小鼠孤雌生殖胚胎分离胚胎干细胞，并研究了其在体内、体外的分化潜能．结果发现:从小鼠孤雌生殖的胚胎中可分离出胚胎干细胞(pES)，可以传代培养25代，能表达很强的碱性磷酸酶，核型稳定呈40XX．培养至第18代的pES在体内可诱导肿瘤形成，并可分化为三个胚层的组织细胞．免疫组化结果显示:神经细胞特异性烯醇化酶（NSE）、肌肉特异性肌动蛋白?-actin均呈阳性，表明分离培养的pES在体内可至少分化为来自外胚层和中胚层的组织细胞．传至第20～24代的pES细胞，经体外定向诱导分化，可定向分化为节律性收缩的心肌细胞及神经细胞．免疫组化检测显示节律性收缩的心肌细胞表达?-actin，而神经细胞表达NSE．结果表明:利用免疫外科法可从孤雌生殖的小鼠胚胎建立pES，这些pES在体内、体外都具有分化为多种类型细胞的潜能．

Isolation and Differentiation Potential of Mouse Parthenogenetic Embryonic Stem Cells

Mouse pES isolated from mouse parthenogenetic blastocysts have the ability to proliferate at least 25 generations in vitro and show very strong activity of alkaline phosphate enzyme.The karyotype of pES was showed as 40XX.To investigate the diffenentiation potential of mouse pES in vivo,pES at the 18 generation were trypsinized and transferred into the testicle envelope of Kunming mouse,histomorphological analysis showed that pES maintained the developmental potential to differentiate into advanced derivatives of all three primary germ layers-ectoderm,endoderm and mesoderm-in vivo.Immuno histochemical results showed that mouse testis had Neuron specific enolase(NSE) and muscle specific α-actin positive cells which derived from pES.Mouse pES at 20 to 24 passages were directionally induced to differentiate in vitro.The results showed that they can be induced into nerve cells and myocardial cells with the rhythm of contraction.Immunocytochemistry showed that myocardial cells were α-actin positive and nerve cells were NSE positive.