Differentiation of human umbilical cord mesenchymal stem cells into dermal fibroblasts in vitro
Differentiation of human umbilical cord mesenchymal stem cells into dermal fibroblasts in vitro
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DOI：
10.1016/j.bbrc.2011.09.001
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发表时间：
2011-10-07
影响因子：
3.1
通讯作者：
Tao, Ran
中科院分区：
文献类型：
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作者：
Han, Yanfu;Chai, Jiake;Tao, Ran
组织源性脐带间充质干细胞（UCMSCs）可以轻易获得，避免伦理或道德约束，并显示出良好的多能性和增殖潜力。UCMSCs被认为是再生医学中有前景的干细胞来源。在本研究中，我们在无菌条件下收集新生儿脐带组织，并通过组织附着法分离UCMSCs。流式细胞术检测UCMSC细胞表面标记物。第三代，在条件诱导培养基中诱导UCMSCs向真皮成纤维细胞分化。采用成纤维细胞特异性单克隆抗体和I型和III型胶原的实时PCR免疫荧光检测诱导结果。UCMSCs表现出成纤维细胞样形态，在原代培养后14至18天达到90%的融合度。培养的UCMSCs显示CD73、CD29、CD44、CD105和hla - 1的阳性染色，但CD34、CD45、CD31和HLA-DR的阳性染色不明显。分化后，诱导细胞的I型胶原、III型成纤维细胞特异性蛋白、波形蛋白、去丝蛋白免疫染色均为强阳性，非诱导细胞的免疫染色为弱或阴性；诱导细胞中I型胶原和ii型胶原mRNA的总转录量高于非诱导细胞。这些结果表明，UCMSCs可以通过条件诱导培养基诱导分化为成纤维细胞，进而可以用作组织工程真皮的种子细胞。(C) 2011爱思唯尔公司版权所有。
Tissue-derived umbilical cord mesenchymal stem cells (UCMSCs) can be readily obtained, avoid ethical or moral constraints, and show excellent pluripotency and proliferation potential. UCMSCs are considered to be a promising source of stem cells in regenerative medicine. In this study, we collected newborn umbilical cord tissue under sterile conditions and isolated UCMSCs through a tissue attachment method. UCMSC cell surface markers were examined using flow cytometry. On the third passage, UCMSCs were induced to differentiate into dermal fibroblasts in conditioned induction media. The induction results were detected using immunofluorescence with a fibroblast-specific monoclonal antibody and real time PCR for type I and type III collagen. UCMSCs exhibited a fibroblast-like morphology and reached 90% confluency 14 to 18 days after primary culture. Cultured UCMSCs showed strong positive staining for CD73, CD29, CD44, CD105, and HLA-I, but not CD34, CD45, CD31, or HLA-DR. After differentiation, immunostaining for collagen type I, type III fibroblast-specific protein, vimentin, and desmin were all strongly positive in induced cells, and staining was weak or negative in non-induced cells; total transcript production of collagen type I and collagen type Ill mRNA was higher in induced cells than in non-induced cells. These results demonstrate that UCMSCs can be induced to differentiate into fibroblasts with conditioned induction media and, in turn, could be used as seed cells for tissue-engineered dermis. (C) 2011 Elsevier Inc. All rights reserved.