摘要
对不同代数去卵巢骨质疏松模型大鼠骨髓间充质干细胞(OVX-rBMSCs)生物学活性进行综合评价,为进一步选取药物筛选的细胞模型提供实验依据。采用全骨髓贴壁筛选法,体外分离、培养OVX-rBMSCs,并通过形态学观察、细胞表面分子标记物(CD29、CD45、CD90)检测、细胞增殖能力检测、多向诱导分化实验(成骨、成脂、成软骨、成神经、成内皮诱导)等技术对P1、P2、P3、P4代OVX-rBMSCs的细胞生物学活性进行综合评价。结果显示,全骨髓贴壁筛选法成功分离培养OVX-rBMSCs,其中P4代细胞呈典型的干细胞形态特征,表面标记物(CD29、CD90)阳性率、细胞增殖指数以及向成骨、成脂、成软骨、成神经、成内皮方向的分化能均优于P1、P2、P3细胞(P<0.05)。结果表明:随着传代次数增加,OVX-rBMSCs纯度及生物学活性指标逐渐优化,其中,P4代均优于其他各代细胞,可作为骨质疏松药物筛选的细胞模型。
This study aimed to comprehensively evaluate the biological activity in different passage populations of mesenchymal stem cells （BMSCs） derived from bone marrow in ovariectomy osteoporotic rats （named OVX-rBMSCs）, providing experimental basis for new osteoporotic drug development and research. OVX-rBMSCs were isolated and cultured in vitro by the whole bone marrow adherent screening method. The morphological observation, cell surface markers （CD29, CD45, CD90） detection, cell proliferation, induced differentiation experimental detection were performed to evaluate the biological activity of Passage 1, 2, 3, 4 populations （P1, P2, P3, P4） OVX-rBMSCs. The results showed that whole bone marrow adherent culture method isolated and differentially subcultured OVX-The morphology of P4 OVX-rBMSCs was identical fibroblast-like and had the characteristics of ultrastructure of stem cells. The CD29 positive cells rate, CD90 positive cells rate, cell proliferation index, and the osteogenic, adipogenic, chondrogenic differentiation capacities of P4 OVX-rBMSCs were significantly better than those of other populations （P 〈 0.05）. OVX-rBMSCs purity and biological activity were gradually optimized with the passaged, and among them P4 cells were superior to all the other populations. Based on these results, we report that the P4 OVX-rBMSCs model developed in this study can be used to develop a new and effective medical method for osteoporotic drug screening.
作者
王彦焱
凌彬
赵志杰
吕鑫
罗丽
龚忠诚
赵春梅
吴江
WANG Yanyan LING Bin ZHAO Zhijie LU Xing LUO Li GONG Zhongcheng ZHAO Chunmei WU Jiang(Department of Rheumatology, The First Affiliated Hospital of Xinjiang Medical University, Uriimqi 830054, China Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China Xuchang City People＇s Hospital, Xuchang 461000, China School of Basic and Forensic Medicine, Sichuan University, Chengdu 610041, China)
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2016年第5期916-922,共7页
Journal of Biomedical Engineering
基金
新疆维吾尔自治区自然科学基金青年基金资助项目(2014211C078)
关键词
骨质疏松
骨髓间充质干细胞
药物筛选
osteoporosis
bone marrow mesenchymal stem cells
drug screening
