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体外特异微环境诱导人胎盘间充质干细胞分化成子宫平滑肌细胞的研究 被引量:5

Simulated uterus microenvironment induced human placental mesenchymal stem cells differentiation to uterus smooth muscle cells in vitro
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摘要 目的体外建立人胎盘间充质干细胞(pMSC)的分离扩增方法,研究其在模拟的特异性微环境中向子宫平滑肌细胞(uSMC)的分化。方法取足月健康新生儿胎盘组织,纯化并扩增pMSC,检测表面标志和并鉴定多向分化潜能。同时原代培养uSMC,改良Trans well培养体系,建立模拟特异性微环境,共培养诱导pMSC向uSMC定向分化。并通过标志性蛋白及细胞功能进行鉴定。结果pMSC同骨髓间充质干细胞一样,具有活跃增殖的能力,表达于细胞标志,具有间充质细胞的特征及多向分化能力。pMSC与uSMC共培养后,其形态逐渐向uSMC形态过渡,并表达uSMC最具特异性的分化晚期标志蛋白肌球蛋白重链(MHC)。诱导获得的细胞表达雌激素受体,并对雌激素的刺激具有反应性的功能变化。结论利用胎盘组织可获取大量pMSC,为组织工程的种子细胞和基因工程的载体细胞提供新的、可行的来源。pMSC的分化具有环境依赖性,在体外模拟的特异性微环境中pMSC可分化为有功能的子宫平滑肌细胞。 Objective To develop a new method to promote the differentiation of mesenchymal stem cells derived from human placenta (pMSC) to uterus smooth muscle cells (uSMC) in simulated uterus microenvironment. Methods MSCs were isolated from human placenta, cultivated, and analyzed for their phenotype by flow cytometry. The muhipotential differentiation of the pMSC was examined by chondrogenic, adipogenic, and osteogenetic induction, uSMC were isolated from uteri resected during operation and cocultivated with the pMSC in a Transwell chamber simulating Two, 4, and 8 days later RT-PCR and Western blotting were used to detect the mRNA and protein expression of α-actin, calmodulin, and myosin heavy chains ( MHC), the markers of smooth muscle differentiation at the early, middle, and late stages. On day 8 RT-PCR was used to detect the expression of estrogen receptor in these 2 groups of cells, then estrogen was used to stimulate these cells and the protein kinase C (PKC) activity was examined. Results The pMSC could be induced into adipocytes, osteocytes, and chondrocytes respectively. After co-culture with uSMC, the morphology of the pMSC changed closely into that of the uSMC, and MHC was expressed in the pMSC. Estrogen receptor was positive in both groups of cells. The PKC activity increased, especially in the cell membrane, after stimulation of estrogen. Conclusion The postpartum human placenta can be used as an important and novel source of muhipotent stem cells for tissue engineering and genetic engineering. Placental MSC have the potential to differentiate into smooth muscle cells under the simulated uterus microenvironment in vitro.
出处 《中华医学杂志》 CAS CSCD 北大核心 2008年第45期3200-3204,共5页 National Medical Journal of China
基金 国家自然科学基金资助项目(30571949) 北京科技新星计划基金资助项目(2006A66)
关键词 间充质干细胞 胎盘 平滑肌 子宫 Mesenchymal stem cells Placenta Smooth muscle Uterus
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参考文献14

  • 1Li CD, Zhang WY, Jiang XX, et al. Human-placenta-derived mesenehymal stem cells inhibit proliferation and function of allogeneie immune cells. Cell Tissue Res, 2007,330:437-446.
  • 2Fukuhara S, Tomita S, Yamashiro S, et al. Direct cell-cell interaction ofcardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro. Thorac Cardiovasc Surg, 2003, 125:1470-1480.
  • 3李长东 ,张为远 ,李荷莲 ,江小霞 ,张毅 ,毛宁 .人胎盘间充质干细胞对脐血淋巴细胞转化的影响[J].中华医学杂志,2005,85(24):1704-1707. 被引量:6
  • 4Pittinger M, Mackay A, Beck S, et al. Muhilineage potential of adult human mesenchymal stem ceils. Science, 1999, 284:143- 147.
  • 5Kawada H, Ando K, Tsuji T, et al. Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system. Exp Hematol, 1999, 27:904-915.
  • 6Miano JM, Cserjesi P, Ligon KL, et al. Smooth muscle myosin heavy chain exelusively marks the smooth musele lineage during mouse embryo genesis. Cire Res, 1994,75:803-812.
  • 7Andrews WV, Hansen JA, Janovick , et al. Gonadotropin releasing hormone modulation of protein kinase C activity I perifused anterior pituitary all cultures. Endocrinology, 1990, 127:2393-2399.
  • 8Bhagavati S. Stem cell based therapy for skeletal muscle diseases. Curt Stern Cell Res Ther, 2008,3:219-228.
  • 9Luciano CA, Anastasios PS, Karl HS, et al . Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction. PNAS ,2005,102 : 11474-11479.
  • 10Stephen GB, Adrian CS, Cay MK. Direct cell contact influences bone marrow mesenchymal stem cell fate. Int J Bioehem Cell Biol, 2004, 36:714-727.

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