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人骨髓间充质干细胞在体外软骨形成分化及去分化过程中表面标志物的变化

Changes in surface markers of human mesenchymal stem cells during the chondrogenic differentiation and dedifferentiation processes in vitro.

作者信息

Lee Hyun Jung, Choi Byung Hyune, Min Byoung-Hyun, Park So Ra

机构信息

Department of Orthopaedic Surgery, School of Medicine, Ajou University, Suwon, South Korea.

出版信息

Arthritis Rheum. 2009 Aug;60(8):2325-32. doi: 10.1002/art.24786.

Abstract

OBJECTIVE

To investigate surface markers showing specific changes during the chondrogenic differentiation and dedifferentiation of human mesenchymal stem cells (MSCs).

METHODS

Human MSCs from adult bone marrow were subjected to chondrogenic differentiation in 3-dimensional (3-D) alginate culture with or without transforming growth factor beta3 (TGFbeta3) for 2 weeks, followed by dedifferentiation in monolayer for 1 week. Surface antigens were selected from those previously reported to show changes in expression during dedifferentiation of human articular chondrocytes (HACs).

RESULTS

Flow cytometry was used to identify 3 groups of surface antigens with differential expression patterns that were quite different from those previously reported on HACs. Two groups of antigens were expressed at high levels on human MSCs. The expression of the first group of antigens (CD44, CD58, CD81, CD90, CD105, and CD166) was decreased reversibly by the 3-D alginate culture and irreversibly in the presence of TGFbeta3, except for CD81, which showed reversible changes regardless of TGFbeta3. The expression of the second group of antigens (CD49c, CD49e, and CD151) was decreased during chondrogenic differentiation only in the presence of TGFbeta3. During all experimental stages, the expression of the third group of antigens (CD14, CD26, CD49f, CD54, CD106, CD119, and CD140a) was maintained at low levels (expressed on <30% of cells), although with some fluctuations.

CONCLUSION

We speculate that the second group of surface antigens could be negative markers for chondrogenic differentiation of human MSCs.

摘要

目的

研究在人骨髓间充质干细胞(MSCs)软骨形成分化和去分化过程中出现特定变化的表面标志物。

方法

将来自成人骨髓的人MSCs在添加或不添加转化生长因子β3(TGFβ3)的三维(3-D)藻酸盐培养体系中进行软骨形成分化2周,随后在单层培养体系中进行去分化1周。表面抗原选自先前报道的在人关节软骨细胞(HACs)去分化过程中表达发生变化的那些抗原。

结果

采用流式细胞术鉴定出3组具有不同表达模式的表面抗原,这些模式与先前报道的HACs上的模式有很大差异。两组抗原在人MSCs上高水平表达。第一组抗原(CD44、CD58、CD81、CD90、CD105和CD166)的表达在3-D藻酸盐培养体系中可逆性降低,在存在TGFβ3时不可逆性降低,但CD81除外,无论有无TGFβ3,其表达均呈现可逆性变化。第二组抗原(CD49c、CD49e和CD151)的表达仅在存在TGFβ3时在软骨形成分化过程中降低。在所有实验阶段,第三组抗原(CD14、CD26、CD49f、CD54, CD106、CD119和CD-140a)的表达维持在低水平(在<30%的细胞上表达),尽管有一些波动。

结论

我们推测第二组表面抗原可能是人MSCs软骨形成分化的阴性标志物。

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