Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Tissue Engineering, 639 Zhi Zao Ju Road, Shanghai 200011, PR China.
Biomaterials. 2010 May;31(13):3564-71. doi: 10.1016/j.biomaterials.2010.01.050. Epub 2010 Feb 12.
Adipose-derived stem cells (ASCs) are considered as a promising cell source for cartilage regeneration. However, the heterogeneity of this cell source may affect their ability in cartilage formation. It is therefore necessary to establish an efficient method for isolating the cells that have chondrogenic potential. To date, no specific markers have been reported to be able to isolate such a cell population from human adipose tissue. In recent studies, endoglin (CD105) has been known as a relatively specific marker for identifying mesenchymal stem cells, but no studies show it is related to chondrogenic potential of human ASCs. In this study, human cells from adipose tissue were isolated, cultured, and sorted according to CD105 expression. The sorted cells were then subjected to adipogenic, osteogenic, and chondrogenic induction to confirm their multi-potentiality. In adipogenic conditions, CD105- cells showed stronger Oil Red staining and higher expression of adipose-specific genes compared to CD105+ cells. By contrast, CD105+ cells exhibited better osteogenic potential with stronger Alizarin Red staining and higher expression of osteogenic specific genes than CD105- cells. Noticeably, CD105+ cells also exhibited a much stronger chondrogenic potential than CD105- cells, with stronger collagen II staining and higher gene expression of collagen II and aggrecan. Most importantly, CD105+ cells could form a homogeneous cartilage-like tissue when seeded into a biodegradable scaffold and cultured in chondrogenic media for 8 weeks. These results indicate that sorting of ASC subpopulation with CD105 as a marker may allow better in vitro chondrogenesis and thus provide an important implications for cartilage regeneration and reconstruction using autologous cells from adipose tissue.
脂肪来源的干细胞(ASCs)被认为是软骨再生有前途的细胞来源。然而,这种细胞来源的异质性可能会影响其在软骨形成中的能力。因此,有必要建立一种有效的方法来分离具有软骨形成潜力的细胞。迄今为止,尚未报道有特定的标记物能够从人脂肪组织中分离出这样的细胞群体。在最近的研究中,内皮糖蛋白(CD105)已被认为是鉴定间充质干细胞的相对特异性标记物,但尚无研究表明其与人类 ASC 的软骨形成潜力有关。在这项研究中,根据 CD105 表达从脂肪组织中分离、培养和分选人细胞。然后将分选的细胞进行成脂、成骨和成软骨诱导,以确认其多能性。在成脂条件下,与 CD105+细胞相比,CD105-细胞的油红染色更强,脂肪特异性基因的表达更高。相比之下,CD105+细胞表现出更好的成骨潜能,其茜素红染色更强,成骨特异性基因的表达高于 CD105-细胞。值得注意的是,与 CD105-细胞相比,CD105+细胞的软骨形成潜力也强得多,其胶原蛋白 II 染色更强,胶原蛋白 II 和聚集蛋白的基因表达更高。最重要的是,当将 CD105+细胞接种到可生物降解的支架中并在软骨形成培养基中培养 8 周时,它们可以形成均匀的软骨样组织。这些结果表明,以 CD105 作为标记物分选 ASC 亚群可能允许更好的体外软骨生成,从而为使用脂肪组织的自体细胞进行软骨再生和重建提供重要意义。