Evaluation of Endothelial Cells Differentiated from Amniotic Fluid-Derived Stem Cells
Evaluation of Endothelial Cells Differentiated from Amniotic Fluid-Derived Stem Cells
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DOI：
10.1089/ten.tea.2011.0392
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发表时间：
2012-06-01
影响因子：
4.1
通讯作者：
Jacot, Jeffrey G.
中科院分区：
文献类型：
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作者：
Benavides, Omar M.;Petsche, Jennifer J.;Jacot, Jeffrey G.
羊水作为干细胞来源有很大的希望，特别是在新生儿应用中，可以分离和使用自体细胞。本研究探讨了化学介导的羊水干细胞（AFSC）向内皮细胞的分化，并验证了AFSC衍生的内皮细胞（AFSC-EC）的功能。AFSC是从妊娠中期羊水中分离出来的，这些羊水是作为双胞胎输血综合征孕妇治疗干预的一部分而从妊娠中期羊水中分离出来的。未分化的AFSC具有正常的核型，具有胚胎干细胞标志物SSEA 4、造血干细胞标志物c-kit和间充质干细胞标志物CD 29、CD 44、CD 73、CD 90和CD 105阳性的细胞亚群。此外，这些细胞对内皮标记物CD 31和造血分化标记物CD 45呈阴性。AFSC在血管内皮生长因子（VEGF）浓度范围为1至100 ng/mL的内皮生长培养基中培养。2周后，AFSC-EC表达血管性血友病因子、内皮型一氧化氮合酶、CD 31、VE-钙粘蛋白和VEGF受体2。此外，表达CD 31的细胞百分比与高达50 ng/mL的VEGF浓度呈正相关，在更高浓度下没有增加。AFSC-EC显示干细胞标志物c-kit和SSEA 4减少，并且在形态上与人脐静脉内皮细胞（HUVEC）相似。在功能测定中，AFSC-EC形成网络并代谢乙酰化低密度脂蛋白，这也是HUVEC的特征。AFSC-EC的硝酸盐水平，一氧化氮合成的间接测量，显着高于未分化的控制，显着低于HUVEC。这些结果表明，AFSC可以分化为功能性内皮样细胞，并可能有潜力为再生医学策略中使用的构建体提供血管化。
Amniotic fluid holds great promise as a stem cell source, especially in neonatal applications where autologous cells can be isolated and used. This study examined chemical-mediated differentiation of amniotic fluid-derived stem cells (AFSC) into endothelial cells and verified the function of AFSC-derived endothelial cells (AFSC-EC). AFSC were isolated from amniotic fluid obtained from second trimester amnioreduction as part of therapeutic intervention from pregnancies affected with twin-twin transfusion syndrome. Undifferentiated AFSC were of normal karyotype with a subpopulation of cells positive for the embryonic stem cell marker SSEA4, hematopoietic stem cell marker c-kit, and mesenchymal stem cell markers CD29, CD44, CD73, CD90, and CD105. Additionally, these cells were negative for the endothelial marker CD31 and hematopoietic differentiation marker CD45. AFSC were cultured in endothelial growth media with concentrations of vascular endothelial growth factor (VEGF) ranging from 1 to 100 ng/mL. After 2 weeks, AFSC-EC expressed von Willebrand factor, endothelial nitric oxide synthase, CD31, VE-cadherin, and VEGF receptor 2. Additionally, the percentage of cells expressing CD31 was positively correlated with VEGF concentration up to 50 ng/mL, with no increase at higher concentrations. AFSC-EC showed a decrease in stem cells markers c-kit and SSEA4 and were morphologically similar to human umbilical vein endothelial cells (HUVEC). In functional assays, AFSC-EC formed networks and metabolized acetylated low-density lipoprotein, also characteristic of HUVEC. Nitrate levels for AFSC-EC, an indirect measure of nitric oxide synthesis, were significantly higher than undifferentiated controls and significantly lower than HUVEC. These results indicate that AFSC can differentiate into functional endothelial-like cells and may have the potential to provide vascularization for constructs used in regenerative medicine strategies.