人孤雌胚胎干细胞与正常胚胎干细胞分化能力的比较  

Comparison of the differentiation capability between human parthenogenetic embryonic stem cells and normal embryonic stem cells

作　　者：欧阳琦[1] 林戈[1,2] 周晓樱[2] 卢光琇[1,2] 

机构地区：[1]中南大学生殖与干细胞工程研究所,长沙410078 [2]人类于细胞国家工程研究中心,长沙410078

出　　处：《解剖学报》2010年第6期785-789,共5页Acta Anatomica Sinica

基　　金：国家863计划基金资助项目(2006AA02A102);国家973计划基金资助项目(2005CB522705)

摘　　要：目的比较人类孤雌胚胎干细胞(pESCs)系与正常胚胎干细胞(nESCs)系的分化能力,探讨pESCs是否和nESCs一样具有多向分化潜能。方法将pESCs和nESCs分别注射到重症联合免疫缺陷(SCID)小鼠体内形成畸胎瘤,瘤体组织切片和HE染色后进行组织学分析;将pESCs和nESCs体外悬浮培养形成拟胚体(EB),利用RT-PCR检测3个胚层主要器官以及滋养层细胞发育关键基因的表达;将pESCs和nESCs定向诱导分化为滋养层细胞,通过流式细胞仪测定人绒毛膜促性腺激素-β(hCG-β)阳性细胞比例以及通过酶联免疫吸附测定(ELISA)进行hCG-β的定量分析。结果在体内生长和体外培养过程中,pESCs和nESCs均能够向3个胚层的细胞类型分化。在SCID小鼠体内可形成畸胎瘤,有神经上皮、软骨、腺上皮等3个胚层的衍生物产生;pESCs和nESCs来源的EB在体外自发分化5～21d后,均检测到3个胚层主要器官以及滋养层细胞发育关键基因的表达;pESCs定向分化为滋养细胞后,可以检测到hCG-β的表达,但其阳性细胞比例和分泌量均低于nESCs。结论 pESCs具有向3个胚层以及滋养层细胞分化的能力,但是向滋养层细胞分化的能力仍低于nESCs。Objective The differentiation capability of human parthenogenetic embryonic stem cells （pESCs） and normal embryonic stem cells （nESCs） was compared to estimate whether the pluripotent potential of pESCs was equal to that of nESCs. Methods pESCs and nESCs were injected into the rear leg of mice with severe combined immunodeficieney disease （SCID） to form teratoma, which was then processed for histological analysis by HE staining; pESCs and nESCs were detached to grow as aggregates in suspension to form embryoid body （EB） , which were collected for the analysis of the key genes expression related with the development of main organs from all three germ layers and trophoblast using RT-PCR. Further, pESCs and nESCs were induced to differentiate into trophoblasts and detected the human chorionic gonadotrophin-β（hCG-β） content in the culture medium by ELISA as well as the percentage of hCG-β positive cells by flow cytometry. Results pESCs and nESCs can differentiate into the cell types from three germ layers both in vivo and in vitro differentiation assay, pESCs and nESCs were able to form teratomas with a complex pattern of differentiation to three germ layers, such as neural epithelia, cartilage and glandular epithelium. The key genes related with the development of main organs from all three germ layers and tropoblast were expressed in day-5 and day-21 EB from pESCs and nESCs, pESCs can be induced to differentiate into trophoblasts and expressed hCG-β, but the content of hCG-β and the percentage of hCG-β positive cells were lower in pESCs than that in nESCs. Conclusion pESCs can differentiate into three germ layers and tropoblast although the differentiation potential to trophoblasts decreased in pESCs while compared with nESCs.

关 键 词：胚胎干细胞 孤雌胚胎干细胞 分化 细胞培养 人 

分 类 号：Q2[生物学—细胞生物学]