全骨髓贴壁法分离培养大鼠骨髓间充质干细胞研究

Isolating and Culturing Rat Bone Marrow Mesenchymal Stem Cells by the Whole Bone Marrow Adherence Method

目的探讨体外全骨髓贴壁法分离大鼠骨髓间充质干细胞的可行性,为体外获取纯度较高的骨髓间充质干细胞提供理论基础。方法采用全骨髓贴壁法分离培养大鼠骨髓间充质干细胞,倒置相差显微镜下观察细胞的黏附、增殖及细胞形态的变化,并采用MTT法绘制P3代细胞生长曲线。结果全骨髓贴壁法分离出的骨髓间充质干细胞经多次换液及传代后得到形态比较一致的典型的纤维样细胞,细胞呈漩涡样、栅栏样及鱼群样集落式生长,细胞间紧密排列,细胞生长曲线呈近似"S"型。结论全骨髓贴壁法能分离出纯度较高的骨髓间充质干细胞,且能最大限度的保持细胞活力。

Objective To explore the feasibility of isolating and culturing rat bone marrow mesenchymal stem cells （BMSCs） by the whole bone marrow adherence method in vitro, and to provide a theoretical basis for obtaining high purity BMSCs. Method Isolating and culturing rat bone marrow mesenchymal stem ceils by the whole bone marrow adher- ence method; observing cell adhesion, proliferation and cell morphology changes by inverted phase contrast microscope; and drawying cell growth curve of P3 generation by MTr assay. Results Bone marrow mesenchymal stem cells were isola- ted by the whole bone marrow adherence method after repeated change medium and passage, and cells presented typical fi- broblast - like morphology, and closely packed like whirlpool, palisade, fish. The P3 cell growth curve was similar to the ＂S＂. Conclusion The whole bone marrow adherence method was able to isolate BMSCs in higher purity, and could main- tain the maximum cell viability.