小鼠颌骨间充质干细胞优化获取及鉴定  

Optimal Isolation and Characterization of Murine Mandible-derived Mesenchymal Stem Cells

作　　者：赵航[1] 盛青[1] 苏俭生[1] 

机构地区：[1]同济大学附属口腔医院修复科,口腔生物医学及转化医学实验室,上海200072

出　　处：《口腔颌面外科杂志》2014年第4期256-260,共5页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81371949);上海市科委重点项目(11nm0504300);上海市科委医学重点项目(13411951201)

摘　　要：目的：摸索优化获取小鼠颌骨间充质干细胞（mesenchymal stem cells,MSCs）的方法。方法 ：通过调整小鼠年龄、胶原酶浓度及酶消化时间等,采用骨片培养法从C57BL/6小鼠下颌骨中分离培养MSCs,并进行生物学鉴定。结果：采用3～4周龄小鼠、0.3%胶原酶消化2 h的方法,分离培养出的细胞增殖能力最强,第3代MSCs即可高表达CD44、CD90及低表达CD34、CD45。各种条件所得MSCs均在成骨诱导后经茜素红染色、成软骨诱导后经甲苯胺蓝染色,结果呈阳性,其成骨、成软骨能力相同。结论：该改良后的骨片培养法可在较短时间内分离出较高纯度的小鼠颌骨间充质干细胞,适用于多种实验研究需要。Objective： To optimize the method of isolation of murine mandible-derived mesenchymal stem cells （MSCs）.Methods： MSCs were harvested and culture-expanded from C57BL/6 murine mandible by digestion of bone chips. The morphological characteristics and colony forming unit-fibroblastic （CFU-F） were observed respectively. MTT assay was used to analyze the proliferation of MSCs. Flow cytometry （FCM） was used for detecting the expression of surface antigens. The potential of MSCs differentiation into osteoblasts and chondrocytes was assessed by histochemical staining. Results： The cultured MSCs of the experiment group （3～4 weeks murines, 0.3% collagenase for 2 hours）, expressed a more rapid proliferation and a higher positive expressions of CD44、CD90 at the early passage 3, while the markers of which were low for CD34、CD45. The corresponding Alizarin red and Toluidine blue staining were positive after induction of osteogenesis and chondrogenesis in all groups. Conclusion： The selected bone chip method is optimal to obtain a high purity of murine mandible MSCs in a relatively short time. The cultured MSCs had a multi-lineage differentiation capacity.

关 键 词：小鼠 颌骨 间充质干细胞 增殖 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]