Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK.
Rheumatology (Oxford). 2013 Oct;52(10):1758-68. doi: 10.1093/rheumatology/ket205. Epub 2013 Jun 26.
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.
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.
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.
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.
我们之前曾报道过，在培养的滑膜间充质干细胞（MSCs）中存在单细胞衍生的克隆细胞群，这些细胞具有不同的分化潜能。本研究旨在探讨用于分离具有不同功能的 MSC 亚群的标记物组合。
采用酶消化法从人滑膜中分离细胞并进行培养扩增。采用流式细胞术分析 MSC 标记物组合对细胞表型进行分析。采用 FACS DiVA 细胞分选仪进行分选。对分选的细胞群体进行克隆形成能力、生长动力学、细胞衰老和软骨-成骨潜能评估。
在培养扩增过程中，CD39 在 CD73(+)细胞群中的共定位鉴定出一个小细胞亚群，该亚群在所有测试供体中从传代 1（P1）至少维持到 P12。CD73(+)CD39(+)细胞亚群 Sox9 和 Runx2 的表达水平更高，软骨-成骨潜能显著大于 CD73(+)CD39(-)细胞亚群。相反，其克隆形成能力和增殖能力较低。分选的 MSC 亚群与原代 MSC 之间的细胞衰老没有差异。值得注意的是，从新鲜滑膜细胞群体中分离出的 CD73(+)CD39(-)和 CD73(+)CD39(+)细胞亚群之间，软骨-成骨潜能没有差异。
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性滑膜 MSC 群体中分离出具有更高软骨-成骨潜能的特定 MSC 亚群。我们预计，这种方法将增强基于细胞的治疗方案在修复骨软骨缺损方面的一致性。
