Pluripotency of Amniotic Fluid-Derived Stem Cells

人类多能干细胞（hPSC）对于再生疗法的发展至关重要，特别是当自体的特化细胞不能以足够的数量获得时。尽管前景广阔，但多能胚胎干细胞（ES）也有一些公认的缺点。这些包括致瘤性和组织相容性匹配的困难。其他类型的hPSC可以克服这些限制。在机构范围的计划公告中，特别指出羊水是hPSC的潜在非胚胎来源。我们最近描述了克隆的人AFS细胞系，并证明它们能够产生包括三个胚胎胚层中的每一个的代表的细胞谱系。其他人已经描述了使用定义的遗传因子将体细胞重编程为ES样状态，称为诱导多能干细胞（iPS）。我们假设AFS细胞代表了比ES和iPS细胞更高级的发育阶段，同时仍然保留了高度的多能性。特别是，从初步数据的支持下，我们预计，AFS细胞将不会产生畸胎瘤的条件下，有利于肿瘤形成的细胞和iPS细胞，因此，关键的比较基因表达的AFS和ES或iPS细胞应导致显着的见解功能的多能性和相关的畸胎瘤形成。这种比较中的一个混淆因素是来自不同人类的细胞系具有遗传多样性。为了精确地关注对应于发育阶段的差异，我们建议比较来自相同个体的AFS和iPS细胞的匹配对。这将通过使用定义的因子将AFS细胞重新编程为更原始的ES样状态来实现。(We指的是这样的重编程细胞iPS-AFS细胞）。在我们原来的R 01应用程序的修订版，我们打算聘请两名来自大学以外的新员工。一个新的初级博士后研究员和一个新的初级技术员。

Human pluripotent stem cells (hPSCs) win be crucial for the development of regenerative therapies, especially when autologous, specialized cells cannot be obtained in sufficient quantity. Despite great .promise, pluripotent embryonic stem (ES) cells have some recognized drawbacks. These include tumorigenicity and difficulty of histocompatibility matching. Other classes of hPSCs may overcome these limitations. In the agency-wide Program Announcement, amniotic fluid is noted specifically as a potential non-embryonic source for hPSCs. We recently described clonal human AFS cell lines and demonstrated that they are able to give rise to cell lineages that include representatives of each of the three embryonic germ layers. Others have described the use of defined genetic factors to reprogram somatic cells to an ES-like state, termed induced pluripotent stem (iPS) cells. We hypothesize that AFS cells represent a developmentally more advanced stage than ES and iPS cells, while nevertheless retaining a high degree of pluripotency. In particular, with support from preliminary data, we anticipate that AFS cells will not give rise to teratomas under conditions conducive to tumor formation by its and iPS cells, Therefore, critical comparison of genes expressed by AFS and ES or iPS cells should lead to significant insights into functions essential for pluripotency and also into those associated with teratoma formation. A confounding factor in such comparisons is that cell lines derived from different human beings are genetically diverse. In order to focus precisely on differences · corresponding to developmental stage, we propose to compare matched pairs of AFS and iPS cells from the same individuals. This will be achieved by using defined factors to reprogram AFS cells to the more primitive ES-like state. (We refer to such reprogrammed cells iPS-AFS cells). In this revised version of our original R01 application we intend to hire two new employees from outside the University. A new junior post-doctoral fellow and a new junior technician.