Placenta Mesenchymal Stem Cell Derived Exosomes Confer Plasticity on Fibroblasts
Placenta Mesenchymal Stem Cell Derived Exosomes Confer Plasticity on Fibroblasts
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DOI：
10.1002/jcb.25459
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发表时间：
2016-07-01
影响因子：
4
通讯作者：
Morita, Ikuo
中科院分区：
文献类型：
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作者：
Tooi, Masayuki;Komaki, Motohiro;Morita, Ikuo
间充质干细胞（MSC）条件培养基（MSC-CM）已被报道，以促进伤口愈合。外泌体含有核酸、蛋白质和脂质，并作为细胞间通讯媒介物用于介导一些旁分泌效应。然而，MSC衍生的外泌体（MSC-exo）的功能仍然难以捉摸。在这项研究中，我们分离人胎盘MSC（PlaMSC）来源的exosomes（PlaMSC-exo），并在体外检测其功能。使用酶消化从人足月胎盘分离PlaMSC。以PlaMSC条件培养液（PlaMSC-CM）为原料，经超离心法制备PlaMSC-exo。通过实时逆转录酶PCR分析（real-time PCR）测量与PlaMSC-exo孵育后正常成人皮肤成纤维细胞（NHDF）中干性相关基因（如OCT 4和NANOG）的表达。使用Oct 4-EGFP报告基因小鼠来源的真皮成纤维细胞评估PlaMSC-exo对OCT 4转录活性的影响。通过碱性磷酸酶（ALP）、茜素红S-和油红O-染色分别评价PlaMSC-exo对NHDF成骨细胞和脂肪细胞分化的促进作用。成骨细胞和脂肪细胞相关基因的表达也通过实时PCR进行了评估。用PlaMSC-exo处理NHDF显著上调OCT 4和NANOG mRNA表达。PlaMSC-exo还增强OCT 4的转录。用PlaMSC-exo处理的NHDF在成骨和成脂诱导培养基中表现出成骨细胞和脂肪细胞分化。PlaMSC-exo可增加成纤维细胞OCT 4和NANOG mRNA的表达。因此，PlaMSC-exo影响成纤维细胞向成骨细胞和脂肪细胞分化的能力。这一结果为MSCs的临床应用提供了新的可能性。(C)2015 Wiley Periodicals，Inc.
Mesenchymal stem cell (MSC)-conditioned medium (MSC-CM) has been reported to enhance wound healing. Exosomes contain nucleic acids, proteins, and lipids, and function as an intercellular communication vehicle for mediating some paracrine effects. However, the function of MSC-derived exosomes (MSC-exo) remains elusive. In this study, we isolated human placenta MSC (PlaMSC)-derived exosomes (PlaMSC-exo) and examined their function in vitro. PlaMSCs were isolated from human term placenta using enzymatic digestion. PlaMSC-exo were prepared from the conditioned medium of PlaMSC (PlaMSC-CM) by ultracentrifugation. The expression of stemness-related genes, such as OCT4 and NANOG, in normal adult human dermal fibroblasts (NHDF) after incubation with PlaMSC-exo was measured by real-time reverse transcriptase PCR analysis (real-time PCR). The effect of PlaMSC-exo on OCT4 transcription activity was assessed using Oct4-EGFP reporter mice-derived dermal fibroblasts. The stimulating effects of PlaMSC-exo on osteoblastic and adipocyte-differentiation of NHDF were evaluated by alkaline phosphatase (ALP), and Alizarin red S- and oil red O-staining, respectively. The expression of osteoblast- and adipocyte-related genes was also assessed by real-time PCR. The treatment of NHDF with PlaMSC-exo significantly upregulated OCT4 and NANOG mRNA expression. PlaMSC-exo also enhanced OCT4 transcription. The NHDF treated with PlaMSC-exo exhibited osteoblastic and adipocyte-differentiation in osteogenic and adipogenic induction media. PlaMSC-exo increase the expression of OCT4 and NANOG mRNA in fibroblasts. As a result, PlaMSC-exo influence the differentiation competence of fibroblasts to both osteoblastic and adipocyte-differentiation. It shows a new feature of MSCs and the possibility of clinical application of MSC-exo. (C) 2015 Wiley Periodicals, Inc.