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作 者:肖龙艳[1] 傅晋翔[1] 张学光[2] 陈秋[2] 张宏[1] 王盼君[1] 孙谕[1] 王明元[3] 古彦铮[2]
机构地区:[1]苏州大学附属第二医院,江苏省苏州市215004 [2]苏州大学生物技术研究所,江苏省苏州市215000 [3]苏州市中心血站,江苏省苏州市215000
出 处:《中国组织工程研究与临床康复》2012年第1期35-38,共4页Journal of Clinical Rehabilitative Tissue Engineering Research
基 金:国家自然科学基金资助项目(81071943)~~
摘 要:背景:骨髓中存在非黏附干细胞,该类细胞形态及表型特征与黏附骨髓间充质干细胞相似。目的:观察胎源性非黏附骨髓基质细胞的基本特性。方法:采用反复贴壁分离法培养胎源性骨髓非黏附基质细胞,每24h取悬浮细胞分瓶培养,共4次;采用MTT法测定第1,3,5代细胞生长曲线,流式细胞仪行细胞周期测定和表面标志分析;并取第1代细胞行造血集落培养,第3代细胞成骨和成脂诱导培养。结果与结论:反复贴壁分离培养4次后仍有细胞贴壁并有集落形成,剩余细胞经培养7d后即可得到原代骨髓非黏附基质细胞。骨髓非黏附基质细胞形态不均匀,高表达整合素及内皮细胞表面标记,中等量表达CD106,同时也表达造血细胞标记CD34;体外可分化为成骨细胞及造血细胞,但成脂分化能力较弱。BACKGROUND: There are non-adhesion stem cells in bone marrow stem cells, and the morphology and phenotype characteristics of the cells are similar to adhesion bone marrow mesenchymal stem cells. OBJECTIVE: To explore the characteristics of fetal non-adherent bone marrow stromal cells (NA-BMSCs). METHODS: The fetal NA-BMSCs were cultivated by the method of adherent separation repeatly ("pull-offs"), the suspended cells were incubated in another culture bottle every 24 hours for 4 times. The growth curve of the passage 1, 3, 5 NA-BMSCs was detected by MTT assay, and the phenotype of the NA-BMSCs was determined with flow cytometry. Passage 1 NA-BMSCs were preformed with hematopoietic colony assay and passage 3 NA-BMSCs were preformed with osteogenic and adipogenic induction. RESULTS AND CONCLUSION: After four times’ "pull-off", there were still asherent cells and colony formation; the resulting cells were continuing to culture for 7 days and those cells were the primary NA-MSCs (P0). The morphology of the cells was uneven; they highly expressed the integrin and the surface markers of endothelial cells with a middle level of CD106. Interestingly, they also expressed the surface markers of hematopoietic cells CD34. Those cells had multi-differentiation potency in vitro and they could differentiate into osteoblasts and hematopoietic cells under different inducing medium, but the capability of lipid differentiation was weak.
关 键 词:非黏附骨髓基质细胞 分化能力 造血干细胞 间充质干细胞 特性
分 类 号:R394.2[医药卫生—医学遗传学]
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