大鼠关节软骨干细胞的分离培养及其特性的鉴定

Isolation,culture and identification of rat articular cartilage stem cells

目的分离培养大鼠关节软骨干细胞(ACSCs)并鉴定其特性。方法运用纤连蛋白黏附法从大鼠关节软骨细胞中分选出ACSCs,流式细胞技术分别鉴定干细胞阳性表面抗原(阳性标志物CD90与CD44)和阴性表面抗原(阴性标志物CD45、CD31与CD34)在ACSCs中的表达水平,单克隆形成实验鉴定ACSCs的单克隆形成能力,成骨、成脂及成软骨诱导鉴定ACSCs的多向分化潜能。结果纤连蛋白可以特异性地分选出ACSCs。ACSCs高表达CD90与CD44,几乎不表达CD45、CD31与CD34。经过7d培养,单个ACSC可以形成大于32个细胞的单克隆细胞团。ACSCs具备很强的成骨、成脂和成软骨分化能力。结论大鼠关节软骨内存在ACSCs,ACSCs具有比较典型的干细胞特征。

Objective To isolate and culture rat articular cartilage stem cells （ACSCs）, and identify their characteristics. Methods ACSCs were isolated from rat articular cartilages by fibronectin-conglutination assay. The expression of positive stem cell surface markers （CD90 and CD44） and negative stem cell surface markers （CD45, CD31 and CD34） in ACSCs was detected by flow cytometry. The single cell colony-forming efficiency of ACSCs was determined by single cell colony- forming assay. The multidirectional differential potential of ACSCs was identified by osteogenesis, adipogenesis and chondrogensis experiment. Results ACSCs were successfully isolated. CD44 and CD90 were highly expressed in ACSCs, while CD31, CD34 and CD45 were not expressed in ACSCs. Seven days after culture, single ACSC formed single cell colony containing more than 32 cells. ACSCs demonstrated a high osteogenesis, adipogenesis and chondrogenesis potential.Conclusion ACSCs are present in rat articular cartilages, and ACSCs have typical characteristics of stem cells.