摘要
目的:研究人骨形态发生蛋白-2(human bone morphogenetic protein-2,hBMP-2)基因转染的兔骨髓基质细胞(bone marrow stromal cells,MSC)与生物活性陶瓷(Bioactive glass ceramics,BGC)体外培养条件下的生物相容性。方法:应用脂质体介导法将携带hBMP-2基因的重组载体PcDNA3-hBMP2导入体外培养的MSC中,G418筛选获得阳性克隆,继续扩增培养。细胞分为实验组(转染的细胞与BGC复合),对照组(单纯转染的细胞),在不同时间用倒置相差显微镜观察。MTT法进行细胞增殖测定,并进行细胞蛋白含量和碱性磷酸酶的定量检测。结果:原位杂交证实基因转染成功,形态特征及生物学行为证实携带hBMP-2基因的MSC无致瘤性,转染的MSC体外培养时复合或不复合BGC均生长良好,BGC利于细胞的贴附、生长与增殖,并对细胞的功能无不良影响。结论:基因转染的MSC可以作为一种新型的种子细胞,BGC是较理想的骨组织工程支架材料,基因转染的MSC复合BGC用于骨缺损的修复,具有广阔的临床应用前景。
Objective:To study the biocompatibility of human bone morphogenetic proteion-2 gene transfected bone marrow stromal celis with biodegradable polymers in vitro culture. Methods:By means of lipofectamine,the cultured MSCs were transfected with PcDNA3- hBMP2. Subsequently, the positive celis were screened out by G418 and the positive clone was collected and expanded. Then the expanded celis were divided into BGC group (in which the celis were cultured with BGC) and transfected celis group (without BGC).The proliferation and differentiation of all the transfected celis were observed at different time point under inverted microscope. Proliferation of the transfected celis were evaluated by MTT assay, and the activity of alkaline phosphatase and the total micro-protein contents in these celis were quantitatively detected. Results: The cultured MSCs were proved to be transfected successfully through hybridization in situ and the transfected celis were proved non-neoplastigenic according to cellular morphology and behavioral biology. The transfected celis grew well in vitro without the presence of BGC, which biological and morphological characteristics were similar to those of normai transfected celis. BGC could improve the adhesion, growth and proliferation of the transfected celis,whereas had no adverse effects on cell functions. Conclusions:The gene transfected MSCs may be a new species of seed celis. And BGC is a kind of optimal scaffold material for bone tissue engineering, vvhich may be used to repair defected bone in clinical treatrnents.
出处
《中国临床解剖学杂志》
CSCD
北大核心
2004年第5期542-545,共4页
Chinese Journal of Clinical Anatomy
