人胚胎干细胞建系的研究现状与存在的问题

人胚胎干细胞系的建立，对人类胚胎发生和人类发育生物学研究、人类新基因的发现和功能研究以及基因治疗、细胞和组织的移植治疗等领域的突破性进展具有重大意义；回顾了人胚胎干细胞建系研究的历程，就建系的几种方案、路线、意义和可行性进行了探讨；详细系统地说明了迄今为止建立人胚胎干细胞系所需要的饲养层类型、培养基组成、添加细胞因子种类及其作用；分析了建立和维持人胚胎干细胞系所需消化酶的种类及其作用以及目前常用的几种传代方法；从若干方面总结了人胚胎干细胞系的鉴定方法，并对建立和维持人胚胎干细胞系中存在的若干问题进行了剖析，提出了目前急待解决的问题。

Current State of Establishing and Maintaining Human Embryonic Stem Cell Lines and Key Problems in These Studies

Derivation of human embryonic stem cell and embryonic germ cell lines has widespread and far reaching significance on human basic research and transplantation therapies.Human pluripotential stem cells provide an exciting new model for studying early human embryogenesis, understanding normal human development and abnormal development,provide a powerful system for discovering human novel genes and testing their function, offer new strategies for discovering of novel growth factors and medicines and promise a renewable source of cells for tissue transplantation, cell replacement and gene therapies. Research history of establishment of human ES and EG cell lines is reviewed. Several methods of establishment of these cell lines involving in the protocal, route, significance and possibility are disscussed. Selection of the feeder layer, medium, and supplemental cytokines and their roles in establishing and maintaining human ES and EG cell lines at present are illustrated in detail systematically.Effects and used methods of several kinds of digestive en zyme in propagations are prepared.Several methods for identifying human ES and EG cells are summarized.At the end, some key problems which are urgent to resolve in these studies at present are put forward and analyzed.