Comparative characterization of CD271+ and CD271- subpopulations of CD34+ human adipose-derived stromal cells
Comparative characterization of CD271+ and CD271- subpopulations of CD34+ human adipose-derived stromal cells
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DOI：
10.1002/jcb.26496
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发表时间：
2018-05-01
影响因子：
4
通讯作者：
Covas, Dimas T.
中科院分区：
文献类型：
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作者：
Beckenkamp, Liziane R.;Souza, Lucas E. B.;Covas, Dimas T.
脂肪来源的基质/干细胞（ASC）是基于细胞的治疗的有希望的候选者。然而，缺乏能够明确识别这些细胞的标记物，来自不同组织的基质祖细胞之间细胞表面分子的差异表达以及由培养引起的细胞改变是需要全面解决的现象，以改善ASC纯化，从而完善我们对它们的功能和治疗效率的认识。在这项研究中，我们研究了用于纯化骨髓间充质干细胞的标记物CD 271在从人脂肪组织的CD 34（+）基质细胞富集ASC中的潜力。基于CD 271表达（CD 45（-）CD 31（-）CD 34（+）CD 271（+）和CD 45（-）CD 31（-）CD 34（+）CD 271（-）细胞）对推定的ASC群体进行分类，并比较其克隆形成效率、增殖、免疫表型特征和多谱系潜力。为了阐明它们的天然身份，我们还询问了新鲜分离的细胞中关键血管周围细胞标志物的表达。CD 271（-）细胞的克隆形成效率是CD 271（+）细胞的2倍。培养后，两个群体的后代显示出相似的免疫表型特征和体外脂肪形成和软骨形成潜力，而CD 271（+）细胞产生更多的钙化细胞外基质。有趣的是，未培养的新鲜分离的CD 271（+）细胞显示出比CD 271（-）细胞更高的周细胞相关标志物的表达，并定位于血管周壁的内部区域。我们的结果表明，具有体外ASC特征的细胞可以从人脂肪组织的CD 271（+）和CD 271（-）基质群体中获得。此外，基因表达谱和原位定位分析表明，CD 271（+）群体显示出周细胞表型。
Adipose-derived stromal/stem cells (ASCs) are promising candidates for cell-based therapies. However, the lack of markers able to unequivocally identify these cells, the differential expression of cell surface molecules among stromal progenitors from different tissues and cellular alterations caused by culture are phenomena that need to be comprehensively addressed in order to improve ASC purification and consequently refine our knowledge about their function and therapeutic efficiency. In this study, we investigated the potential of CD271, a marker used for purification of bone marrow-derived mesenchymal stem cells, on enriching ASCs from CD34(+) stromal cells of human adipose tissue. Putative ASC populations were sorted based on CD271 expression (CD45(-)CD31(-)CD34(+)CD271(+) and CD45(-)CD31(-)CD34(+)CD271(-) cells) and compared regarding their clonogenic efficiency, proliferation, immunophenotypic profile, and multilineage potential. To shed light on their native identity, we also interrogated the expression of key perivascular cell markers in freshly isolated cells. CD271(-) cells displayed twofold higher clonogenic efficiency than CD271(+) cells. Upon culture, the progeny of both populations displayed similar immunophenotypic profile and in vitro adipogenic and chondrogenic potentials, while CD271(+) cells produced more calcified extracellular matrix. Interestingly, uncultured freshly isolated CD271(+) cells displayed higher expression of pericyte-associated markers than CD271(-) cells and localized in the inner region of the perivascular wall. Our results demonstrate that cells with in vitro ASC traits can be obtained from both CD271(+) and CD271(-) stromal populations of human adipose tissue. In addition, gene expression profiling and in situ localization analyses indicate that the CD271(+) population displays a pericytic phenotype.