人脐血间充质干细胞与β-磷酸三钙的生物相容性

Biocompatibility of human umbilical cord blood derived mesenchymal stem cells and beta-tricalcium phosphate

背景:体内实验显示,β-磷酸三钙多孔陶瓷是较为理想的骨组织工程支架材料,但由于体内植入实验受多种因素的影响,不能很好反映细胞的生长、增殖和表型变化。目的:观察体外人脐血间充质干细胞与β-磷酸三钙多孔陶瓷的生物相容性。方法:将培养的第6代人脐血间充质干细胞悬液滴注入β-磷酸三钙内部进行复合,然后将干细胞-支架材料复合物置入含体积分数为10%胎牛血清的α-MEM培养体系中培养,于培养第4,8,12天电镜下观察人脐血间充质干细胞在材料表面及内部生长情况,采用MTT测试法绘制细胞生长曲线,并进行DNA含量、蛋白质含量测定。结果与结论:人脐血间充质干细胞与β-磷酸三钙体外复合后能够在β-磷酸三钙支架材料表面及内部的孔隙内贴附,且生长良好,其DNA复制和蛋白合成功能不受β-磷酸三钙的影响。说明人脐血间充质干细胞和β-磷酸三钙支架材料生物相容性良好,二者可作为种子细胞和支架材料用于组织工程化骨与软骨的构建。

BACKGROUND： Experiments in vivo has showed that β-tricalcium phosphate （β-TCP） porous ceramic is an ideal bone tissue engineering scaffold material, but it cannot reflect growth, proliferation, and phenotypic changes of cells well, due to implantation of experiments in vivo was affected by various factors. OBJECTIVE： To investigate the biocompatibility of human umbilical cord blood derived mesenchymal stem cells （hUCB-MSCs） in vitro and β-TCP porous ceramic. METHODS： The 6th passage hUCB-MSCs suspension were dripped into β-TCP to undergo combination.The compound of stem cells and scaffold materials was transferred into the culture medium comprising α-minimum essential medium （α-MEM） supplemented with 10% fetal bovine serum （FBS）. The surface of materials and internal growth status of hUCB-MSCs was observed under scanning electron microscope （SEM） at 4, 8, and 12 days. MTT assay was used to draw curve of cell growth and the DNA and protein content were determined. RESULTS AND CONCLUSION： HUCB-MSCs can attach in the surface and internal pores of β-TCP scaffold materials after hUCB-MSCs combined with β-TCP, and its growth is good, which DNA replication and protein synthesis function were not affected by β-TCP. It suggested that hUCB-MSCs and β-TCP scaffold materials have good biocompatibility, and which can be used as seed cells and scaffold materials for tissue engineering bone and cartilage construction.