STRO-1阳性与阴性骨髓间充质干细胞成骨能力的对比  

Comparison of osteogenic potential in STRO-1 positive and negative bone marrow mesenchymal stem cells

作　　者：周万林 何斌 申雄成 黄坤 田仁元 袁野 黄文良 Zhou Wanlin;He Bin;Shen Xiongcheng;Huang Kun;Tian Renyuan;Yuan Ye;Huang Wenliang(Department of Orthopedics,Third Affiliated Hospital of Zunyi Medical University(First People’s Hospital of Zunyi City),Zunyi 563000,Guizhou Province,China;Department of Orthopedics,Honghuagang District People’s Hospital of Zunyi City,Zunyi 563000,Guizhou Province,China;Department of Orthopedics,Moutai Hospital of Guizhou Province,Renhuai 564500,Guizhou Province,China)

机构地区：[1]遵义医科大学第三附属医院(遵义市第一人民医院)骨科,贵州省遵义市563000 [2]遵义市红花岗区人民医院骨科,贵州省遵义市563000 [3]贵州省茅台医院骨科,贵州省仁怀市564500

出　　处：《中国组织工程研究》2025年第36期7719-7727,共9页Chinese Journal of Tissue Engineering Research

基　　金：贵州省科技计划项目(黔科合基础-ZK(2021)一般387),项目负责人:黄文良;贵州省科技计划项目(黔科合成果-LC[2024]019),项目负责人:黄文良;贵州省卫生健康委科学技术基金项目(gzwjk2024-410),项目负责人:申雄成;贵州省卫生健康委科学技术基金项目(gzwjk2020-1-131),项目负责人:何斌。

摘　　要：背景:随着年龄增长,骨髓间充质干细胞的再生能力和分化功能逐渐下降,导致骨组织修复效果减弱。因此,探索成骨潜能更高的骨髓间充质干细胞亚群对骨组织工程的发展具有重要意义。目的:评估STRO-1阳性和阴性骨髓间充质干细胞在成骨诱导培养条件下的成骨分化能力差异。方法:分离并培养SD大鼠骨髓间充质干细胞,采用流式细胞术和免疫荧光鉴定CD29、CD45、CD90和STRO-1的表达;利用免疫磁细胞分选技术分离STRO-1阳性和阴性骨髓间充质干细胞,将两组细胞分别进行成骨诱导7,14 d,采用qRT-PCR和Western blot分析两组细胞成骨相关基因(Ⅰ型胶原、Runt相关转录因子2、骨保护素、骨钙素)及蛋白表达水平差异,采用茜素红染色和碱性磷酸酶染色观察钙结节形成情况。结果与结论:流式细胞术显示CD29和CD90高表达,CD45低表达,STRO-1的阳性表达率为12.8%,免疫荧光结果与流式细胞术一致。磁细胞分选后发现STRO-1阳性细胞的集落形成率高于STRO-1阴性细胞;STRO-1阳性细胞在第14天的成骨分化能力显著高于第7天,且在第14天时成骨相关标志物的表达水平显著高于STRO-1阴性细胞,差异有显著性意义(P<0.01)。结果表明:STRO-1阳性骨髓间充质干细胞在成骨能力上具有显著优势,为骨组织工程中理想种子细胞的筛选提供了理论依据,未来在骨缺损修复中有望成为一种潜在的治疗手段。BACKGROUND:With aging,the regenerative capacity and differentiation function of bone marrow mesenchymal stem cells progressively decline,reducing bone tissue repair efficacy.Thus,identifying bone marrow mesenchymal stem cell subpopulations with enhanced osteogenic potential is of significant importance for advancing bone tissue engineering.OBJECTIVE:To evaluate the osteogenic differentiation potential differences between STRO-1 positive and negative bone marrow mesenchymal stem cells under osteogenic induction conditions.METHODS:SD rat bone marrow mesenchymal stem cells were isolated and cultured.The expression of CD29,CD45,CD90,and STRO-1 was identified via flow cytometry and immunofluorescence.Immunomagnetic cell sorting was used to separate STRO-1 positive and negative bone marrow mesenchymal stem cells.The cells of two groups were subjected to osteogenic induction for 7 and 14 days.qRT-PCR and western blotting were performed to analyze differences in osteogenesis-related gene expression(Collagen I,Runt-related transcription factor 2,osteoprotegerin,and osteocalcin)and protein levels.Alizarin red staining and alkaline phosphatase staining were used to observe calcium nodule formation.RESULTS AND CONCLUSION:Flow cytometry showed high expression levels of CD29 and CD90 and low expression of CD45,with a positive STRO-1 expression rate of 12.8%.Immunofluorescence results were consistent with those of flow cytometry.After magnetic cell sorting,STRO-1 positive cells demonstrated a higher colony formation rate than STRO-1 negative cells.On day 14,STRO-1 positive cells showed significantly higher osteogenic differentiation potential than on day 7,with significantly elevated osteogenesis-related marker levels compared to STRO-1 negative cells(P<0.01).The findings indicate that STRO-1 positive bone marrow mesenchymal stem cells exhibit significant advantages in osteogenic potential,providing a theoretical basis for their selection as ideal seed cells in bone tissue engineering.In future applications,they may represent a

关 键 词：STRO-1 骨髓间充质干细胞 成骨分化 免疫磁细胞分选 骨缺损修复 种子细胞 骨组织工程 工程化干细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R681.7