Identification of subpopulations with characteristics of mesenchymal progenitor cells from human osteoarthritic cartilage using triple staining for cell surface markers.
Identification of subpopulations with characteristics of mesenchymal progenitor cells from human osteoarthritic cartilage using triple staining for cell surface markers.
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DOI：
10.1186/ar1210
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发表时间：
2004
影响因子：
4.9
通讯作者：
Brenner RE
中科院分区：
文献类型：
--
作者：
Fickert S;Fiedler J;Brenner RE
我们首先使用荧光激活细胞分选（FACS）鉴定并分离了骨关节炎软骨中具有间充质祖细胞（MPCs）特征的细胞亚群。通过酶促分离骨关节炎软骨细胞，并直接或在经过几代培养扩增后，使用已在人MPCs （CD9, CD44, CD54, CD90, CD166）上鉴定的各种表面标记组合进行FACS分析。结合培养扩增细胞和最初分类的CD9+、CD90+、CD166+细胞衍生的亚群，使用既定的分化方案测试其成骨、成脂和成软骨潜能。通过免疫组织化学和RT-PCR分析分化情况，检测谱系相关标记基因的表达。通过FACS分析，我们发现骨关节炎软骨内CD9、CD44、CD54、CD90和CD166阳性细胞的各种三组组合占总数的2-12%。经黏附培养后，其相对含量明显增高，在24% ~ 48%之间。培养扩增细胞结合后，初步分类的CD9/CD90/CD166三阳性亚群具有成软骨、成骨和成脂肪分化的多能性。总之，人骨关节炎软骨含有具有MPCs特征的细胞。它们在体外培养过程中的相对富集以及细胞分选获得更均匀群体的能力为骨关节炎关节内再生过程激活的未来研究提供了有趣的视角。
We first identified and isolated cellular subpopulations with characteristics of mesenchymal progenitor cells (MPCs) in osteoarthritic cartilage using fluorescence-activated cell sorting (FACS). Cells from osteoarthritic cartilage were enzymatically isolated and analyzed directly or after culture expansion over several passages by FACS using various combinations of surface markers that have been identified on human MPCs (CD9, CD44, CD54, CD90, CD166). Culture expanded cells combined and the subpopulation derived from initially sorted CD9+, CD90+, CD166+ cells were tested for their osteogenic, adipogenic and chondrogenic potential using established differentiation protocols. The differentiation was analyzed by immunohistochemistry and by RT-PCR for the expression of lineage related marker genes. Using FACS analysis we found that various triple combinations of CD9, CD44, CD54, CD90 and CD166 positive cells within osteoarthritic cartilage account for 2–12% of the total population. After adhesion and cultivation their relative amount was markedly higher, with levels between 24% and 48%. Culture expanded cells combined and the initially sorted CD9/CD90/CD166 triple positive subpopulation had multipotency for chondrogenic, osteogenic and adipogenic differentiation. In conclusion, human osteoarthritic cartilage contains cells with characteristics of MPCs. Their relative enrichment during in vitro cultivation and the ability of cell sorting to obtain more homogeneous populations offer interesting perspectives for future studies on the activation of regenerative processes within osteoarthritic joints.
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影响因子：
--
作者：
Jones, EA;Kinsey, SE;McGonagle, D
通讯作者：
McGonagle, D
影响因子：
--
作者：
Barbero, A;Ploegert, S;Martin, I
通讯作者：
Martin, I
影响因子：
4
作者：
Gronthos, S;Zannettino, ACW;Simmons, PJ
通讯作者：
Simmons, PJ
影响因子：
20.3
作者：
Kuçi, S;Wessels, JT;Handgretinger, R
通讯作者：
Handgretinger, R
DOI：
10.1186/ar148
发表时间：
2001
期刊：
Arthritis research
影响因子：
--
作者：
Sandell LJ;Aigner T
通讯作者：
Aigner T