Fraga Ana Maria, de Araújo Erica Sara Souza, Stabellini Raquel, Vergani Naja, Pereira Lygia V
National Laboratory for Embryonic Stem Cells, Dept. Genetics and Evolutionary Biology, University of São Paulo, São Paulo, SP, Brazil.
Methods Mol Biol. 2012;873:1-12. doi: 10.1007/978-1-61779-794-1_1.
Although since 1998 more than 1,200 different hESC lines have been established worldwide, there is still a recognized interest in the establishment of new lines of hESC, particularly from HLA types and ethnic groups underrepresented among the currently available lines. The methodology of hESC derivation has evolved significantly since the initial derivations using human LIF (hLIF) for maintenance of pluripotency. However, there are still a number of alternative strategies for the different steps involved in establishing a new line of hESC. We have analyzed the different strategies/parameters used between 1998 and 2010 for the derivation of the 375 hESC lines able to form teratomas in immunocompromised mice deposited in two international stem cell registries. Here we describe some trends in the methodology for establishing hESC lines, discussing the developments in the field. Nevertheless, we describe a much greater heterogeneity of strategies for hESCs derivation than what is used for murine ESC lines, indicating that optimum conditions have not been identified yet, and thus, hESC establishment is still an evolving field of research.
尽管自1998年以来全球已建立了1200多种不同的人胚胎干细胞（hESC）系，但人们仍对建立新的hESC系感兴趣，特别是那些在现有细胞系中代表性不足的HLA类型和种族群体的细胞系。自最初使用人白血病抑制因子（hLIF）维持多能性进行hESC诱导以来，hESC诱导方法已发生了显著演变。然而，在建立新的hESC系的不同步骤中仍有许多替代策略。我们分析了1998年至2010年间用于诱导375个hESC系的不同策略/参数，这些细胞系能够在免疫缺陷小鼠中形成畸胎瘤，并已存入两个国际干细胞登记处。在此，我们描述了建立hESC系方法的一些趋势，并讨论了该领域的发展。尽管如此，我们发现hESC诱导策略的异质性比用于小鼠胚胎干细胞（mESC）系的策略要大得多，这表明尚未确定最佳条件，因此，hESC的建立仍是一个不断发展的研究领域。
