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作 者:魏利成[1] 刘丹平[2] 蒲勤[3] 王朝燕[1] 蒋清[1] 尹建文[1] 张正[1]
机构地区:[1]湖南省邵阳市第一人民医院骨科,422000 [2]锦州医学院附属第一医院骨科 [3]第四军医大学生化教研室
出 处:《中国骨与关节损伤杂志》2009年第1期52-54,共3页Chinese Journal of Bone and Joint Injury
基 金:国家自然基金资助项目(30672125)
摘 要:目的观察pcDNA3-hBMP2转染兔骨髓基质细胞前后的生长特性变化,为骨组织工程的种子细胞选择打下基础。方法抽取成年雄性新西兰大白兔髂骨骨髓,密度梯度离心获得骨髓基质细胞,培养传至第5代,按处理方法分为常规培养液组(A组),pcDNA3-hBMP2转染组(B组)。观察原代MSC及第5代的细胞形态等生长特性并与转染组进行比较。结果原代MSCs为短梭形,簇形生长,传代细胞呈长梭形,旋涡形生长。转染组细胞72 h后表达BMP2,瞬间表达为100%。转染细胞未经筛选培养4周细胞形态发生较明显地变化,细胞生长周期改变。结论pcDNA3-hBMP2成功转染兔骨髓基质细胞并改变其生物学特性。Objective To observe main biological characteristic and changes before and after MSCs were transfected with pcDNA3 - hBMP2. Methods MSCs from an adult New Zealand white rabbit were isolated and cultivated. MSCs were divided into two groups. Group A was transfected with pcDNA3 - hBMP2 and group B was not transfected as control group. Immunohistochemical staining was used for detecting BMP2 expressing 2 days after transfection. The cell morphological changes, cell divide index and cell cycle were observed after transfection. Results Rabbit MSCs could be successfully transfected by pcDNA3 - hBMP2 through medium fugene 6 and express BMP2. Primary MSCs proliferated as short spindle shape, while post - transfected MSCs showed triangle- shape, and transletted- cells had significant difference in cell cycle and cell divide index. Conclusion Rabbit MSCs can be successfully transfected by pcDNA3 - hBMP2 and after transfection many biological characteristics are significantly changed in this study.
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