Prospective purification of a subpopulation of human synovial mesenchymal stem cells with enhanced chondro-osteogenic potency
Prospective purification of a subpopulation of human synovial mesenchymal stem cells with enhanced chondro-osteogenic potency
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DOI：
10.1093/rheumatology/ket205
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发表时间：
2013-10-01
期刊：
影响因子：
5.5
通讯作者：
De Bari, Cosimo
中科院分区：
文献类型：
--
作者：
Gullo, Francesca;De Bari, Cosimo
客观的。我们之前报道过，在来自人类滑膜的培养间充质干细胞（MSC）中，具有不同分化能力的单细胞衍生克隆细胞群共存。本研究的目的是研究用于前瞻性纯化功能不同的 MSC 子集的标记集。方法。细胞从人滑膜中酶促释放并扩增培养物。使用 MSC 标记组合通过流式细胞术进行表型分析。使用FACS DiVA细胞分选仪进行分选。评估分选的细胞群的克隆形成性、生长动力学、细胞衰老和软骨成骨效力。结果。在培养扩增过程中，CD39 在 CD73(+) 细胞群中的共定位确定了一个小细胞亚群，该亚群在所有测试的供体中从第 1 代 (P1) 一直维持到至少 P12。 CD73(+)CD39(+)细胞亚群比CD73(+)CD39(-)细胞亚群表现出更高的Sox9和Runx2表达水平以及显着更强的软骨成骨效力。相反，它的克隆性和增殖性较低。分选的 MSC 亚群与亲代 MSC 之间的细胞衰老没有差异。值得注意的是，从新鲜滑膜细胞群中纯化的CD73(+)CD39(-)和CD73(+)CD39(+)细胞亚群之间的软骨成骨效力没有可检测到的差异。结论。我们的研究结果表明，CD73 和 CD39 的组合允许从培养扩增的异质滑膜 MSC 群体中前瞻性纯化具有更大软骨成骨效力的独特 MSC 子集。我们预计这种方法将增强基于细胞的骨软骨缺陷修复治疗方案的一致性。
Objective. We previously reported the coexistence, within cultured mesenchymal stem cells (MSCs) from human synovial membrane, of single-cell-derived clonal cell populations with distinct differentiation potency. The aim of this study was to investigate marker sets for prospective purification of functionally distinct MSC subsets.Methods. Cells were enzymatically released from human synovium and culture expanded. Phenotype analysis was performed by flow cytometry using combinations of MSC markers. Sorting was carried out using the FACS DiVA cell sorter. Sorted cell populations were assessed for clonogenicity, kinetics of growth, cell senescence and chondro-osteogenic potency.Results. During culture expansion, the co-localization of CD39 within the CD73(+) cell population identified a small cell subset that was maintained from passage 1 (P1) up to at least P12 in all donors tested. The CD73(+)CD39(+) cell subset displayed higher expression levels of Sox9 and Runx2 and a significantly greater chondro-osteogenic potency than the CD73(+)CD39(-) cell subset. In contrast, it was less clonogenic and proliferative. There was no difference in cell senescence between the sorted MSC subsets and the parental MSCs. Notably, there were no detectable differences in chondro-osteogenic potency between the CD73(+)CD39(-) and CD73(+)CD39(+) cell subsets purified from fresh synovial cell populations.Conclusion. Our findings indicate that the combination of CD73 and CD39 allows the prospective purification from culture-expanded heterogeneous synovial MSC populations of a distinct MSC subset with greater chondro-osteogenic potency. We anticipate that such an approach will enhance the consistency of cell-based therapeutic protocols for the repair of osteochondral defects.