Direct selection of human bone marrow mesenchymal stem cells using an anti-CD49a antibody reveals their CD45med,low phenotype
Direct selection of human bone marrow mesenchymal stem cells using an anti-CD49a antibody reveals their CD45med,low phenotype
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DOI：
10.1046/j.1365-2141.2003.04469.x
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发表时间：
2003-08-01
影响因子：
6.5
通讯作者：
Herve, P
中科院分区：
文献类型：
--
作者：
Deschaseaux, F;Gindraux, F;Herve, P
人骨髓间充质干细胞（MSC）通过成纤维细胞集落形成单元（CFU-F）产生成骨软骨细胞以及脂肪细胞和间充质细胞。到目前为止，这些干细胞是通过细胞培养方法间接分离出来的，并且表型为CD45阴性，而体内的对应物尚不确定。我们的目的是开发一种直接选择方法，并确定以这种方式分离的MSC的表型。采用流式细胞术或磁珠法，用抗cd49a和/或抗cd45抗体选择间充质细胞。所有CFU-F总是在cd49a阳性细胞的小群体中检测到。这些CFU保留了它们的分化潜能，并产生成骨软骨细胞、脂肪细胞和间充质细胞。对未培养细胞的表型研究显示CD45（低），CD34（低），HLA-II细胞群。流式细胞术细胞分选显示，仅从CD49a(+) /CD45（med,low）群体中获得具有CFU-F电位的MSC。此外，在培养时，它们清晰地变成CD45(-), CD34(-), HLA-II-, CD49a（+）。这些结果证实了磁珠可以在不改变骨髓分化潜能的情况下，直接从人骨髓中选择间充质干细胞。这些细胞轻度表达造血标志物CD45，经体外培养后CD45显著下调。因此，CD45与CD49a偶联的表达能够直接选择MSC。
Human bone marrow mesenchymal stem cells (MSC) generate, via a fibroblast colony-forming unit (CFU-F), osteo-chondroblastic cells as well as adipocytes and stromacytes. To date, these stem cells are isolated indirectly using a cell culture method and phenotyped as CD45 negative while the in vivo counterparts are undetermined. Our aim was to develop a direct selection method and to determine the phenotype of the MSC isolated in this way. Mesenchymal cells were selected with anti-CD49a and/or anti-CD45 antibodies using either flow cytometry or a magnetic beads method. All CFU-F were always detected in the small population of CD49a-positive cells. These CFU retained their differentiation potential and gave rise to osteo-chondroblastic cells, adipocytes and stromacytes. Phenotypic studies on uncultured cells revealed a CD45(med,low) , CD34(low) , HLA-II- cell population. Flow cytometry cell sorting showed that MSC with CFU-F potential were obtained only from a CD49a(+) /CD45(med,low) population. In addition, when cultured, they clearly became CD45(-) , CD34(-) , HLA-II- , CD49a(+) . These results confirmed that MSC can be directly selected easily from human bone marrow using magnetic beads without altering their differentiation potential. These cells expressed mildly the haematopoietic marker CD45, which was dramatically downregulated by in vitro culture. The expression of CD45 coupled to CD49a thus enabled direct selection of the MSC.