Umbilical cord-derived mesenchymal stem cells isolated by a novel explantation technique can differentiate into functional endothelial cells and promote revascularization.
Umbilical cord-derived mesenchymal stem cells isolated by a novel explantation technique can differentiate into functional endothelial cells and promote revascularization.
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DOI：
10.1089/scd.2009.0321
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发表时间：
2010-02
影响因子：
4
通讯作者：
Yan Xu;H. Meng;Chang-hong Li;M. Hao;Ya-fei Wang;Zhen Yu;Qian Li;Junling Han;Q. Zhai;L. Qiu
中科院分区：
文献类型：
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作者：
Yan Xu;H. Meng;Chang-hong Li;M. Hao;Ya-fei Wang;Zhen Yu;Qian Li;Junling Han;Q. Zhai;L. Qiu
干细胞移植通过功能血运重建治疗缺血性疾病具有很大的前景。脐带间充质干细胞（UC-MSCs）也是基于细胞的生物工程的理想候选者。在此，我们报告了一种简单有效的分离UC-MSCs的方案的发展，并证实了它们在体外和体内的内皮潜力。采用新颖的外植技术分离UC-MSCs，诱导其向内皮样细胞分化。然后将UC-MSCs移植到缺血小鼠模型中，并在3D凝胶/MMT-CS复合支架上培养。形态学和增殖评估表明，在相对较短的培养时间内，外植体技术可以产生足够的UC-MSCs。内皮特异性标记物（KDR和vWF）和功能标记物（ac-LDL摄取和UEA-1结合）的表达增加，表明体外培养9天后诱导了功能性内皮祖细胞。在缺血后肢小鼠模型中，MSCs移植后4周的缺血/非缺血肢体灌注比达到0.84 +/- 0.09。与假注射组相比，大鼠毛细血管密度增大2.57倍（P < 0.05）。免疫荧光和免疫组织学分析表明，MSCs可能通过融入局部脉管系统来挽救缺血组织。在体外，我们观察到UC-MSCs与3D凝胶/MMT-CS复合支架结合，分泌细胞外基质，保持活力，并保持其增殖能力。综上所述，通过新颖而简单的外植技术分离的UC-MSCs非常适合用于开发基于干细胞的新型血管重建疗法。
Stem cells transplantation holds great promise for the treatment of ischemic diseases through functional revascularization. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are also an ideal candidate for cell-based bioengineering. Herein, we report on the development of a simple and effective protocol to isolate UC-MSCs, and confirm their endothelial potential both in vitro and in vivo. UC-MSCs were isolated by a novel explantation technique and induced to differentiate into endothelial-like cells. Then UC-MSCs were transplanted into ischemic mouse model and cultured on 3D gel/MMT-CS composite scaffolds. Morphological and proliferation assessments show that sufficient UC-MSCs can be generated during a relatively short culture period with explantation technique. Increased expression of endothelial-specific markers (KDR and vWF), and functional markers (ac-LDL uptake and UEA-1 binding), indicate that functional endothelial progenitor cells are induced after 9 days of in vitro culture. In an ischemic hindlimb mouse model, the ratio of ischemic/nonischemic limb perfusion 4 weeks after MSCs transplantation reached 0.84 +/- 0.09. The capillary density of this group was 2.57-fold greater than that of sham-injected mice (P < 0.05). Immunofluorescence and immunohistological analyses indicate that MSCs may act to salvage the ischemic tissue by incorporating into the local vasculature. In vitro, UC-MSCs were observed to incorporate into 3D gel/MMT-CS composite scaffolds, to secrete extracellular matrix, to remain viable, and to retain their proliferation capacity. In conclusion, UC-MSCs isolated by novel yet simple explantation technique are well suited for application in the development of novel stem cell-based revascularization therapies.