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人脂肪干细胞膜片的构建及其生物学特性初探 被引量:3

Preliminary study in construction and biological characteristics of human adipose-derived stem cells sheet
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摘要 目的利用分离纯化后的人脂肪干细胞(adipose-derived stem cells,ADSCs)构建ADSCs膜片,并探讨人ADSCs膜片的生物学特性。方法收集年轻女性腹部手术皮下脂肪组织,用酶消化法成功分离纯化人ADSCs,进行流式细胞学鉴定,使用含有活性因子的完全培养基诱导构建人ADSCs膜片后,扫描电镜观察细胞膜片组织形态,并采用q RT-PCR和ELISA检测人脂肪干细胞膜片相关促生长细胞因子及干性基因的表达水平。结果成功分离人ADSCs,流式细胞学鉴定发现其间充质干细胞表面标记分子CD29、CD44、CD90、CD105均为阳性,造血干细胞表面标记CD45分子为阴性,符合ADSCs表面分子表达特点;人ADSCs进行成脂和成骨诱导后,油红O与茜素红染色阳性,提示人ADSCs具有多向分化潜能;扫描电镜观察膜片中的细胞呈网格样叠加生长,细胞之间连接紧密;qRT-PCR检测结果显示人ADSCs膜片中TGF-β1、FGF2、HGF、VEGF、Col 1A1等促生长细胞因子和OCT4、NANOG等干性基因显著高表达。ELISA检测结果显示人ADSCs膜片上清液高表达促生长细胞因子TGF-β1、FGF、HGF、VEGF。结论我们成功利用人ADSCs诱导成人细胞膜片,该膜片可分泌更高含量的促生长细胞因子并表达更高水平的干性基因,为人ADSCs膜片应用于组织损伤修复提供了重要理论依据。 Objective To construct human adipose-derived stem cells (ADSCs) sheet by isolated and purified ADSCs and explore its biological characteristics. Methods Fatty samples were collected from young women's abdomen. Then human ADSCs were obtained by isolating and purifying the samples by collagenase digestion, and then identified using flow cytometry. Human ADSCs were incubated and cultured in complete medium and formed cell-sheet. The morphology and structure of cells sheets were observed by scanning electron microscope (SEM) and the expressions of growth factors and stem-related genes in cell-sheets were detected by qRT-PCR and ELISA. Results Human ADSCs were cultured successfully in vitro, and mesenchymal stem cells markers including CD29, CD44, CD90, CD105 were positively expressed on human ADSCs while CD45 (one of hematopoietic stem cells markers) was negative, which was consistent with ADSCs' characteristics. After osteogenic and adipogenic induction, the calcified nodule and fat drops could be seen using Alizarin red staining and Oil red staining respectively, which suggested that the human ADSCs had multiple differentiation potential. SEM analysis revealed that highly adherent human ADSCs grew in dense layers. The mRNA level of growth factors including TGF- [3 1, FGF, HGF and VEGF and the expression of stem-related genes like NANOG and OCT4 increased significantly in ADSCs sheets compared with the human ADSCs itself. In addition, the protein level of TGF-β1, FGF2, HGF, VEGF were significantly higher in the cell suspension of cell-sheet by ELISA. Conclusion Human ADSCs cell-sheet is constructed successfully in our study. It can express higher level of some growth factors and stem-related markers, which may provide important evidence for the tissue regeneration and repair using human ADSCs cell-sheet.
出处 《解放军医学院学报》 CAS 2018年第2期144-149,共6页 Academic Journal of Chinese PLA Medical School
基金 陕西省统筹创新项目(2013KTCL03-07)~~
关键词 人脂肪干细胞 细胞膜片 组织再生 human adipose-derived stem cells cell-sheet tissue regeneration
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  • 1管峰,杨利国,程瑞禾,曹少先.骨形态发生蛋白家族及其受体在生殖调控中的作用[J].生理科学进展,2004,35(3):262-264. 被引量:12
  • 2郭永红,罗金燕.TGF-β超家族与Smad信号转导研究进展[J].医学综述,2005,11(8):685-688. 被引量:24
  • 3Uglow EB,Slater S,Sala-Newby GB,et al.Dismantling of cadherin-mediated cell-cell contacts modulates smooth muscle cell proliferation[J].Circ Res,2003,92(12):1314-1321. 被引量:1
  • 4He W,Myers MR,Hanney B,et al.Potent quinoxaline-based inhibitors of PDGF receptor tyrosine kinase activity.Part 2: the synthesis and biological activities of RPR127963 an orally bioavailable inhibitor[J].Bioorg Med Chem Lett,2003,13(18):3097-3100. 被引量:1
  • 5Chung IM,Ueno H,Pak YK,et al.Catheter-based adenovirus-mediated local intravascular gene delivery of a soluble TGF-beta type II receptor using an infiltrator in porcine coronary arteries: efficacy and complications[J].Exp Mol Med,2002,34(4):299-307. 被引量:1
  • 6Schmidt A,Gopfert C,Vlodavsky I,et al.Induction of a hypertrophic growth status of coronary smooth muscle cells is associated with an overexpression of TGF-beta[J].Eur J Cell Biol,2002,81(3):138-144. 被引量:1
  • 7Francis DJ,Parish CR,McGarry M,et al.Blockade of vascular smooth muscle cell proliferation and intimal thickening after balloon injury by the sulfated oligosaccharide PI-88: phosphomannopentaose sulfate directly binds FGF-2,blocks cellular signaling,and i 被引量:1
  • 8Swanson N,Hogrefe K,Javed Q,et al.Vascular Endothelial Growth Factor (VEGF)-Eluting Stents: In Vivo Effects on Thrombosis,Endothelialization and Intimal Hyperplasia.J Invasive Cardiol[J],2003,15(12):688-692. 被引量:1
  • 9Bayes-Genis A,Schwartz RS,Bale LK,et al.Effects of insulin-like growth factor-I on cultured human coronary artery smooth muscle cells[J].Growth Horm IGF Res,2003,13(5):246-253. 被引量:1
  • 10Jeng AY.Utility of endothelin-converting enzyme inhibitors for the treatment of cardiovascular diseases[J].Curr Opin Investig Drugs,2003,4(9):1076-1081. 被引量:1

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