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诱导多能干细胞来源的条件培养基对人脂肪来源干细胞增殖和抗凋亡的影响。

Effects of induced pluripotent stem cells-derived conditioned medium on the proliferation and anti-apoptosis of human adipose-derived stem cells.

作者信息

Lian Rui-Ling, Guo Xiao-Ling, Chen Jian-Su, Guo Yong-Long, Zheng Jia-Fu, Chen Yuan-Wen

机构信息

Department of Ophthalmology, First Affiliated Hospital of Jinan University, Guangzhou, 510632, Guangdong, China.

Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, 510632, Guangdong, China.

出版信息

Mol Cell Biochem. 2016 Feb;413(1-2):69-85. doi: 10.1007/s11010-015-2640-7. Epub 2016 Jan 2.

Abstract

Human adipose-derived stem cells (hASCs) become an appealing source for regenerative medicine. However, with the multi-passage or cryopreservation for large-scale growth procedures in terms of preclinical and clinical purposes, hASCs often reveal defective cell viability, which is a major obstacle for cell therapy. In our study, the effects of induced pluripotent stem cells-derived conditioned medium (iPS-CM) on the proliferation and anti-apoptosis in hASCs were investigated. hASCs at passage 1 were identified by the analysis of typical surface antigens with flow cytometry assay and adipogenic and osteogenic differentiation. The effect of iPS-CM on the proliferation in hASCs was analyzed by cell cycle assay and Ki67/P27 quantitative polymerase chain reaction analysis. The effect of iPS-CM on the anti-apoptosis of hASCs irradiated by 468 J/m(2) of ultraviolet C was investigated by annexin v/propidium iodide analysis, mitochondrial membrane potential assay, intracellular reactive oxygen species assay, Western blotting and caspase activity assays. The effect of iPS-CM on the surface antigen expressions of hASCs was analyzed using flow cytometry assay. The levels of Activin A and bFGF in culture supernatant of hASCs with different treatments were also detected by enzyme-linked immunosorbent assay. iPS-CM promoted proliferation and inhibited apoptosis of hASCs. This discovery demonstrates that iPS-CM might be used as one of the available means to overcome the propagation obstacle for hASCs and make for large-scale growth procedures in terms of preclinical and clinical purposes.

摘要

人脂肪来源干细胞(hASCs)成为再生医学中一个有吸引力的细胞来源。然而,在临床前和临床应用的大规模培养过程中,经过多次传代或冷冻保存后,hASCs常常表现出细胞活力缺陷,这是细胞治疗的一个主要障碍。在我们的研究中,研究了诱导多能干细胞来源的条件培养基(iPS-CM)对hASCs增殖和抗凋亡的影响。通过流式细胞术分析典型表面抗原以及成脂和成骨分化,鉴定第1代hASCs。通过细胞周期分析和Ki67/P27定量聚合酶链反应分析,分析iPS-CM对hASCs增殖的影响。通过膜联蛋白V/碘化丙啶分析、线粒体膜电位检测、细胞内活性氧检测、蛋白质免疫印迹和半胱天冬酶活性检测,研究iPS-CM对经468 J/m(2) 紫外线C照射的hASCs抗凋亡的影响。使用流式细胞术分析iPS-CM对hASCs表面抗原表达的影响。还通过酶联免疫吸附测定法检测不同处理的hASCs培养上清液中激活素A和碱性成纤维细胞生长因子的水平。iPS-CM促进hASCs的增殖并抑制其凋亡。这一发现表明,iPS-CM可能是克服hASCs增殖障碍并有助于临床前和临床应用大规模培养的可用方法之一。

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