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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 the changes of growth characteristics before and after transfection of pcDNA3-hBMP2 into rabbit bone marrow stromal cells, and lay a foundation for the selection of seed cells for bone tissue engineering. Methods Bone marrow stromal cells of adult male New Zealand white rabbits were collected and density-gradient centrifugation was used to obtain bone marrow stromal cells. The cells were passaged for 5 passages and divided into routine culture medium (group A), pcDNA3-hBMP2 transfection group (group B) . The growth characteristics of the primary MSC and the fifth generation of cells were observed and compared with the transfection group. Results The primary MSCs were short fusiform and clustered, and the passage cells were long fusiform and swirling. The cells were transfected 72h after expression of BMP2, transient expression was 100%. Transfected cells cultured without screening for 4 weeks more obvious changes in cell morphology, cell growth cycle changes. Conclusion pcDNA3-hBMP2 transfected rabbit bone marrow stromal cells and change their biological characteristics.