Section of Internal Medicine and Endocrine and Metabolic Sciences (Di.M.I.), Department of Internal Medicine, University of Perugia, Perugia, Italy.
Tissue Eng Part A. 2011 Nov;17(21-22):2651-61. doi: 10.1089/ten.TEA.2010.0587. Epub 2011 Sep 6.
We have developed a simple and rapid method for isolation of human umbilical cord matrix stem cells (hUCMS). The umbilical cord contains a virtual inexhaustible source of adult stem cells. We have substantially modified, simplified, and improved previously reported hUCMS isolation procedures in terms of either used enzyme type, or digestion time, and substantially enhanced the final product yield and purity. The isolated hUCMS were positive for CD90, CD117, and SCF, and negative for CD31 and CD45 surface markers. mRNA and related proteins (i.e., Sox2, Oct4a, Nanog, ABCG2, and c-Myc) that coincide with an uncommitted cell status also were detected. hUCMS express genes and proteins for CD90 and Nestin that are associated with mesenchymal stem cells, as well as other genes that specifically relate to different embryonic germ layers, namely, Vimentin, Sox7, Sox17, FoxA2, E-cadherin, and N-cadherin. hUCMS showed multilineage cell differentiation potential into adipogenic, osteogenic, and neural cell phenotypes, under the influence of lineage-specific, differentiation culture media. Moreover, the basal expression of endocrine cell markers makes these cells seemingly suitable for endocrine cell phenotype differentiation. Noteworthy, Activin A induced hUCMS to acquire definitive endoderm cell markers.
我们开发了一种简单而快速的方法来分离人脐带基质干细胞（hUCMS）。脐带包含了几乎用之不竭的成人干细胞来源。我们在使用的酶类型或消化时间方面对先前报道的 hUCMS 分离程序进行了实质性的修改、简化和改进，并且大大提高了最终产物的产量和纯度。分离的 hUCMS 对 CD90、CD117 和 SCF 呈阳性，对 CD31 和 CD45 表面标志物呈阴性。还检测到与未分化细胞状态一致的 mRNA 和相关蛋白（即 Sox2、Oct4a、Nanog、ABCG2 和 c-Myc）。hUCMS 表达与间充质干细胞相关的 CD90 和 Nestin 的基因和蛋白，以及与不同胚胎胚层特异性相关的其他基因，即 Vimentin、Sox7、Sox17、FoxA2、E-cadherin 和 N-cadherin。在特定谱系分化培养基的影响下，hUCMS 显示出向成脂、成骨和成神经细胞表型的多能性分化潜力。此外，内分泌细胞标志物的基础表达使得这些细胞似乎适合内分泌细胞表型分化。值得注意的是，激活素 A 诱导 hUCMS 获得确定的内胚层细胞标志物。
