SD大鼠骨髓间充质干细胞体外分离培养的实验研究

目的 建立一种简便有效的体外分离纯化及培养扩增大鼠骨髓间充质干细胞(BMSCs)的方法,研究BMSCs的生物学特性,为后续细胞移植和基因治疗提供理想的种子细胞.方法 全骨髓贴壁培养法分离大鼠BMSCs,体外培养和连续传代,在倒置相差显微镜下连续观察细胞的形态变化,利用MTT法测定其生长曲线,流式细胞仪鉴定其表面抗原,成...

Isolation and culture of SD rats' bone marrow-derived mesenchymal stem cells

Objective To establish a simple and effective method of isolation,purification and culture of rat bone marrow-derived mesenchymal stem cells in vitro,and explore their biological characteristics in order to provide an ideal seed cell for stem cells transplantation and gene therapy.Methods BMSCs were isolated by the attachment culture method.They were cultured and passaged continuously,their morphological changes were observed under an inverted phase contrast microscope.Their growth curves were measured by MTT method,and their membrane antigens were detected by flow cytometry.Osteoblastic differentiation and adipocytic differentiation were induced in order to study their potential of multiple differentiation.Results The morphology of BMSCs was identical shuttle-like or polygon-like and the cells showed active proliferative ability.CD29 and CD90 antigen expression was positive in BMSCs,while CD34 and CD45 antigen expression was negative,assayed by flow cytometry.The cells showed their potential of multiple differentiations after induced.Conclusion The attachment culture method can be used to isolate,expand and purify BMSCs in vitro.Cultured BMSCs show stable and rapid growth and have general biological characteristics of mesenchymal stem cells,which lay solid foundation for gene-modified BMSCs and their transplantation in vivo.