人骨髓基质干细胞体外三维立体培养的实验研究

Culture of human bone marrow mesenchymal stem cells by three-dimensional scaffold in vitro: an experimental study

目的:探讨人骨髓基质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)在部分脱钙骨三维支架上立体培养的可行性。方法:将自人骨髓中分离出的骨髓基质干细胞进行培养、扩增,接种于多孔的部分脱钙骨支架上,通过荧光活性染料DiI标记和扫描电镜观察骨髓基质干细胞在部分脱钙骨上的生长和粘附情况,并测量hBMSCs在部分脱钙骨上的粘附率。结果:hBMSCs在部分脱钙骨上的粘附率为(99.1±1)%;DiI标记的细胞于接种后第2、4、7天激光共聚焦检测,见支架空隙中细胞逐渐增多;电镜观察,接种7天后,电镜下见细胞覆盖了部分脱钙骨表面;自部分脱钙骨中份切开,横断面电镜观察见断面中充满细胞。结论:hBMSCs在部分脱钙骨支架的三维立体环境中生长良好,是一种较好的体外培养方法。

Objective： To study the possibility of culture of human bone marrow mesenchymal stem cells by three-dimensional scalffold. Method： Bone marrow mesenchymal stem cells were isolated from human bone marrow and cultured and muhiplified, then seeded in the porous partially demineralized bone matrix. The growth and adhesion were observed by DiI label and scanning electromicroscope. Result： About （99.1 ± 1 ） % of the cells adhered to the porous partially demineralized bone matrix; on the 2nd, 4th and 7th day after the DiI labeled cells were seeded on the scaffold, they were observed under confocal laser scanning microscope , the pore was covered by the cells on the 7th day. On the 7th day, the cells covered the surface of the scaffold; after the scaffold was cut into two parts, it can be observed that the pore in the transverse surface was also filled with cells. Conclusion： Human bone marrow mesenchymal stem cells grow well in the three-dimensional environment of partially demineralized bone matrix, which is a good method for culture of human bone marrow mesenchymal stem cells in vitro.