Bucay Nathan, Yebra Mayra, Cirulli Vincenzo, Afrikanova Ivka, Kaido Thomas, Hayek Alberto, Montgomery Anthony M P
Whittier Institute, Department of Pediatrics, University of California, San Diego, La Jolla, 92037, USA.
Stem Cells. 2009 Jan;27(1):68-77. doi: 10.1634/stemcells.2007-1018.
Using human embryonic stem cells (hESCs), we describe a novel method for the rapid derivation and enrichment of cells that are comparable to primordial germ cells (PGCs) and Sertoli cells. The methodology described is based on modest changes to the growth conditions commonly used to expand hESCs and does not require genetic manipulation or complex three-dimensional culture. Remarkably, we have determined that simply reducing the size of cultured ESC colonies and manipulating the number of feeding cycles, results in the rapid emergence of cells that are comparable to migratory PGCs. Importantly, these cells can be monitored and purified on the basis of the expression of the chemokine receptor CXCR4. Under more stringent differentiating conditions these cells mature and upregulate the expression of specific germ cell markers. Importantly, this process is accompanied by the development of Sertoli-like support cells. Such cells normally provide trophic support and immunoprotection to developing germ cells and may have significant clinical utility in the prevention of graft rejection. The putative Sertoli-germ cell cocultures generated in this study may ultimately be developed to study and manipulate interactions and processes involved in human gametogenesis.
利用人类胚胎干细胞（hESCs），我们描述了一种快速获得和富集与原始生殖细胞（PGCs）及支持细胞相当的细胞的新方法。所描述的方法基于对常用于扩增hESCs的生长条件进行适度改变，且不需要基因操作或复杂的三维培养。值得注意的是，我们已确定，简单地减小培养的ESC集落大小并控制传代次数，会导致快速出现与迁移的PGCs相当的细胞。重要的是，这些细胞可根据趋化因子受体CXCR4的表达进行监测和纯化。在更严格的分化条件下，这些细胞成熟并上调特定生殖细胞标志物的表达。重要的是，这一过程伴随着类支持细胞的发育。此类细胞通常为发育中的生殖细胞提供营养支持和免疫保护，在预防移植物排斥方面可能具有重要的临床应用价值。本研究中产生的假定支持细胞-生殖细胞共培养物最终可能被用于研究和操控人类配子发生过程中涉及的相互作用和过程。
