Comprehensive Phenotypic Characterization of Human Adipose-Derived Stromal/Stem Cells and Their Subsets by a High Throughput Technology
Comprehensive Phenotypic Characterization of Human Adipose-Derived Stromal/Stem Cells and Their Subsets by a High Throughput Technology
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DOI：
10.1089/scd.2012.0346
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发表时间：
2013-01-01
影响因子：
4
通讯作者：
Schubert, Ralf
中科院分区：
文献类型：
--
作者：
Baer, Patrick C.;Kuci, Selim;Schubert, Ralf
由于缺乏明确和独特的细胞标志物，脂肪源性基质/干细胞（ASC）的表征仍然很困难。因此，标记物的组合对于识别细胞是必要的。没有全面的分析免疫表型的扩大塑料贴壁ASCs已公布。因此，本研究的目的是表征培养的ASCs的一般表型，并进一步分析细胞亚群。从接受美容吸脂术的患者的脂肪抽吸物中分离ASC，并在标准细胞培养物中培养。用含有242种抗体和同种型对照的BD Lyoplate（TM）人细胞表面标记筛选板进行综合表型表征。培养的ASCs不仅表现出间充质干细胞（MSC）的特征性表达谱，而且还显示了49种其他标志物表达的供体特异性变异。我们进一步检测了荧光强度散射的标记物，表明亚群具有不同的表达谱。因此，在用抗CD 73、CD 90、CD 105和CD 34、CD 140 b、CD 200、CD 201或CD 36的直接标记抗体染色细胞后进行多色流式细胞术分析，以验证所选的ASC亚群。我们没有检测到CD 34-CD 36双阳性群体，但检测到CD 34（+）-CD 36（-）和CD 34（-）CD 36（+）亚群，这两种亚群均对3种主要MSC标志物CD 73、CD 90和CD 105呈阳性。所有其他检测到的亚群也共表达3种主要MSC标志物，因此满足定义培养的MSC的最低表型标准。我们的研究证明了ASC的第一个全面的表型特征，并清楚地强调了ASC制剂中供体特异性的变异性。
The characterization of adipose-derived stromal/stem cells (ASCs) remains difficult due to the lack of a definitive and unique cellular marker. Therefore, a combination of markers is necessary to identify the cells. No comprehensive analysis of the immunophenotype of expanded plastic adherent ASCs has been published. Therefore, the aim of this study was to characterize the general phenotype of cultured ASCs and to further analyze cellular subsets. ASCs were isolated from lipoaspirates from patients undergoing cosmetic liposuction and cultured in standard cell culture. A comprehensive phenotype characterization was done with the BD Lyoplate (TM) Human Cell Surface Marker Screening Panel containing 242 antibodies and isotype controls. Cultured ASCs not only showed the characteristic expression profile of mesenchymal stem cells (MSCs), but also revealed donor-specific variability in the expression of 49 other markers. We further detected markers with a scattering in the fluorescence intensity, indicating subpopulations with different expression profiles. Therefore, a multi-color flow cytometric analysis was done after staining the cells with direct-labeled antibodies against CD73, CD90, CD105, and either CD34, CD140b, CD200, CD201, or CD36 to verify the selected subpopulations of ASCs. We detected no CD34-CD36 double-positive population, but CD34(+)-CD36(-) and CD34(-)CD36(+) subpopulations, both of which are positive for the 3 main MSC markers, CD73, CD90, and CD105. All other detected subpopulations also co-expressed the 3 main MSC markers, and therefore fulfill the minimal phenotypic criteria for the definition of cultured MSCs. Our study demonstrates the first comprehensive phenotypic characterization of ASCs and clearly highlights donor-specific variability in ASC preparations.