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In Vitro Expansion and Characterization of Mesenchymal Stromal Cells from Peritoneal Dialysis Effluent in a Human Protein Medium

机译:人蛋白质培养基中腹膜透析流出物的间充质基质细胞的体外扩增和鉴定

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摘要

The therapeutic potential of mesenchymal stromal cells (MSCs) from various tissue origins have extensively been explored in both experimental and clinical studies, and peritoneal dialysis effluent-derived MSC (pMSC) may be an easily obtainable MSC source for clinical applications. In this study, we expanded and characterized the pMSCs after expansion in a human protein culture medium. The pMSCs were expanded in plastic dishes with the human protein medium. MSC marker expression was examined by flow cytometry. Spherical formation was tested by hanging drop method, and osteogenic, adipogenic, and chondrogenic differentiation capacities were confirmed by positive staining with Alizarin red, Oil red O, and Alcian blue, respectively. Here, we showed that after four passages of culturing in plastic dishes, pMSCs in the human protein medium displayed a homogeneous pattern of classical MSC markers (positive: CD29, CD44, CD73, CD90, and CD166; negative: CD14, CD34, CD45, CD79a, CD105, CD146, CD271, HLA-DR, SSEA-4, and Stro-1), while in the standard medium, pMSCs from some donors were CD45 or HLA-DR positive. For nonclassical MSC markers, pMSCs were CD200 positive from all the donors, negative for CD163, CD271, CD36, and CD248, and either positive or negative for CD274 and CD140b. Further, pMSCs from the human protein medium had the spherical formation capacity and multipotent differentiation capacity in vitro. In conclusion, upon expansion in a human protein medium, pMSCs showed a differential MSC marker expression profile from those of bone marrow or adipose tissue-derived MSCs and could maintain the multipotency. The therapeutic potential of the pMSCs requires further investigation.
机译:在实验和临床研究中都广泛探索了来自各种组织起源的间充质基质细胞(MSC)的治疗潜力,并且腹膜透析流出物衍生的MSC(pMSC)可能是易于获得的MSC来源,用于临床应用。在这项研究中,我们在人蛋白质培养基中扩增后扩增并表征了pMSC。用人蛋白培养基在塑料皿中扩增pMSC。通过流式细胞术检查MSC标志物的表达。通过悬滴法测试球状形成,并通过分别用茜素红,油红O和阿尔辛蓝阳性染色证实成骨,成脂和成软骨分化能力。在这里,我们展示了在塑料培养皿中培养四次后,人蛋白质培养基中的pMSCs表现出均匀的经典MSC标记模式(阳性:CD29,CD44,CD73,CD90和CD166;阴性:CD14,CD34,CD45, CD79a,CD105,CD146,CD271,HLA-DR,SSEA-4和Stro-1),而在标准培养基中,来自某些供体的pMSCs为CD45或HLA-DR阳性。对于非经典MSC标记,所有供体的pMSC均为CD200阳性,CD163,CD271,CD36和CD248均为阴性,而CD274和CD140b均为阳性或阴性。此外,来自人蛋白培养基的pMSC在体外具有球形形成能力和多能分化能力。总之,在人蛋白培养基中扩增后,pMSCs与骨髓或脂肪组织来源的MSCs的MSC标记物表达谱存在差异,并且可以保持多能性。 pMSCs的治疗潜力需要进一步研究。

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