Manipal Institute of Regenerative Medicine, Manipal University, No 10, Service Road, Domlur Layout, Bangalore 560071, India.
Placenta. 2012 Oct;33(10):850-8. doi: 10.1016/j.placenta.2012.07.001. Epub 2012 Jul 25.
One of the most pressing problems in injury is wound healing and blood vessel formation. The amniotic membrane is important in clinical applications as it is pro-angiogenic, anti-fibrotic and anti-scarring and has low immunogenicity. In this study, we characterized amniotic membrane mesenchymal stem cells (AMMSCs) by their trademark mesenchymal stem cell (MSC) signature and profiled for embryonic pluripotency markers namely alkaline phosphatase, Oct4, Sox2, Nanog, SSEA3 and 4, and Klf4 by RT-PCR and nuclear localization of Oct4 and Nanog by immunocytochemistry. The amnion, although avascular, contains pro-angiogenic factors such as type I, III, IV and V collagen, laminin, and fibronectin in the extra cellular matrix. We, therefore, hypothesized that AMMSCs is pro-angiogenic. Thus, we demonstrate that MSCs derived from the amnion have a natural ability to initiate endothelialization and angiogenesis in vitro. Our results using a wound scratch assay and angiogenesis on Matrigel suggest a pro-angiogenic property of AMMSCs. We also show that native, uninduced AMMSCs are able to form endothelial rings in Matrigel. Further evidence was provided by RT-PCR showing the expression of pro-angiogenic factors such as Tie2, Ang1, VEGF, VEGFR, vWF, KDR and Flt4 in native AMMSCs. Taken together, our results suggest that MSCs from an avascular amnion have an inherent propensity for promoting angiogenesis and could be an ideal choice in wound healing, stroke and ischemic diseases that require rapid vascularization and tissue restoration.
在损伤中，最紧迫的问题之一是伤口愈合和血管形成。羊膜在临床应用中很重要，因为它具有促血管生成、抗纤维化和抗瘢痕形成的作用，且免疫原性低。在这项研究中，我们通过其标志性的间充质干细胞 (MSC) 特征对羊膜间充质干细胞 (AMMSC) 进行了表征，并通过 RT-PCR 对胚胎多能性标志物碱性磷酸酶、Oct4、Sox2、Nanog、SSEA3 和 4 以及 Klf4 进行了分析，通过免疫细胞化学对核定位的 Oct4 和 Nanog 进行了分析。尽管羊膜没有血管，但在细胞外基质中含有促血管生成因子，如 I 型、III 型、IV 型和 V 型胶原、层粘连蛋白和纤维连接蛋白。因此，我们假设 AMMSC 具有促血管生成作用。因此，我们证明了源自羊膜的 MSC 具有在体外启动内皮化和血管生成的天然能力。我们使用划痕实验和 Matrigel 上的血管生成的结果表明 AMMSC 具有促血管生成特性。我们还表明，天然的、未诱导的 AMMSC 能够在 Matrigel 中形成内皮环。通过 RT-PCR 显示促血管生成因子如 Tie2、Ang1、VEGF、VEGFR、vWF、KDR 和 Flt4 在天然 AMMSC 中的表达，进一步提供了证据。综上所述，我们的结果表明，源自无血管羊膜的 MSC 具有促进血管生成的固有倾向，在需要快速血管化和组织修复的伤口愈合、中风和缺血性疾病中可能是理想的选择。
