Isolation of functionally distinct mesenchymal stem cell subsets using antibodies against CD56, CD271, and mesenchymal stem cell antigen-1
Isolation of functionally distinct mesenchymal stem cell subsets using antibodies against CD56, CD271, and mesenchymal stem cell antigen-1
复制标题
DOI：
10.3324/haematol.13740
复制
发表时间：
2009-02-01
期刊：
影响因子：
--
通讯作者：
Buehring, Hans-Joerg
中科院分区：
文献类型：
--
作者：
Battula, Venkata Lokesh;Treml, Sabrina;Buehring, Hans-Joerg
背景通常，间充质干细胞是根据其在塑料表面的黏附能力而从原代组织中分离出来的。这一分离过程受到共培养的造血细胞和/或其他无关细胞的不可预测的影响和/或在去除不需要的细胞过程中消除晚期黏附的间充质干细胞亚群的影响。为了绕过这些限制，已经开发了几种抗体来促进骨髓间充质干细胞的预期分离。最近，我们报道了一组对骨髓间充质干细胞具有良好选择性的单抗，包括针对人骨髓间充质干细胞抗原-1(MSCA-1)的单抗W8B2和针对自然杀伤细胞不表达的CD56表位的39D5。设计与方法用一大组抗CD271、MSCA-1和CD56表面抗原的抗体对健康供者的骨髓间充质干细胞进行分析和分离。集落形成单位成纤维细胞(CFU-F)检测间充质干细胞的生长情况。结果多色细胞分选和CFU-F分析显示，间充质干细胞在MSCA-1(+)CD56(-)组分中的分化倍数为90倍，在MSCA-1(+)CD56(+)组分中的分化倍数为180倍。表型分析显示CD10、CD26、CD106和CD146仅限于MSCA-1(+)CD56(-)间充质干细胞亚群，CD166仅限于MSCA-1(+)CD56(+/-)间充质干细胞亚群。进一步分化发现，软骨细胞和类胰岛主要来源于MSCA-1(+)CD56(+/-)细胞，而脂肪细胞仅来源于MSCA-1(+)CD56(-)细胞。单细胞分选MSCA-1(+)CD56(+)细胞培养可获得表型异质克隆，具有明显的增殖和分化能力。结论获得了具有不同表型和功能的间充质干细胞亚群。我们的数据表明MSCA-1(+)CD56(+)亚群是一个有吸引力的自体软骨细胞移植的起始群体。
BackgroundConventionally, mesenchymal stem cells are functionally isolated from primary tissue based on their capacity to adhere to a plastic surface. This isolation procedure is hampered by the unpredictable influence of co-cultured hematopoietic and/or other unrelated cells and/or by the elimination of a late adhering mesenchymal stem cells subset during removal of undesired cells. To circumvent these limitations, several antibodies have been developed to facilitate the prospective isolation of mesenchymal stem cells. Recently, we described a panel of monoclonal antibodies with superior selectivity for mesenchymal stem cells, including the monoclonal antibodies W8B2 against human mesenchymal stem cell antigen-1 (MSCA-1) and 39D5 against a CD56 epitope, which is not expressed on natural killer cells.Designand Methods Bone marrow derived mesenchymal stem cells from healthy donors were analyzed and isolated by Bow cytometry using a large panel of antibodies against surface antigens including CD271, MSCA-1, and CD56. The growth of mesenchymal stem cells was monitored by colony formation unit fibroblast (CFU-F) assays. The differentiation of mesenchymal stem cells into defined lineages was induced by culture in appropriate media and verified by immunostaining.ResultsMulticolor cell sorting and CFU-F assays showed that mesenchymal stem cells were similar to 90-fold enriched in the MSCA-1(+)CD56(-) fraction and similar to 180-fold in the MSCA-1(+)CD56(+) fraction. Phenotype analysis revealed that the expression of CD10, CD26, CD106, and CD146 was restricted to the MSCA-1(+)CD56(-) mesenchymal stem cells subset and CD166 to MSCA-1(+)CD56(+/-) mesenchymal stem cells. Further differentiation of these subsets showed that chondrocytes and pancreatic-like islets were predominandy derived from MSCA-1(+)CD56(+/-) cells whereas adipocytes emerged exclusively from MSCA-1(+)CD56(-) cells. The culture of single-sorted MSCA-1(+)CD56(+) cells resulted in the appearance of phenotypically heterogeneous clones with distinct proliferation and differentiation capacities.ConclusionsNovel mesenchymal stem cells subsets with distinct phenotypic and functional properties were identified. Our data suggest that the MSCA-1(+)CD56(+) subset is an attractive starting population for autologous chondrocyte transplantation.