接种浓度对人骨髓基质干细胞在脱钙骨上生长和分布的影响

Influence of seeding concentration on the growth and distribution of human bone marrow stem cell on the demineralized bone matrix

目的 研究不同接种浓度对人骨髓基质干细胞在脱钙骨支架上生长过程及分布的影响。方法 荧光活性染料DiI标记人骨髓基质干细胞,接种于部分脱钙骨支架材料上,测定细胞的粘附率;将细胞悬液浓度分别调整为20×106、10×106、5×106/mL和空白组,依次接种于同一块脱钙骨支架4个相邻象限的中心位置上。接种后第2、7 d在倒置荧光显微镜下观察材料表面的细胞分布、生长的情况,测量细胞材料面积比。结果 DiI标记的骨髓基质干细胞的粘附率为(99.9±0.2)％,未标记的骨髓基质干细胞粘附率为(99.1±1)％,两者无统计学差异(P>0.05)。不同浓度细胞的细胞材料面积比无差异(P>0.05)。结论 通过DiI标记可以动态观察细胞在材料上的生长状况。20×106、10×106、5×106/mL接种浓度均能达到覆盖脱钙骨表面直接接种部位的目的。

Objective To investigate the influence of different cell seeding density on the distribution and growth of human bone marrow stem cells( BMSCs) on partially demineralized bone matrix( pDBM). Methods human BMSCs were labeled with Dil and seeded on the pDBM . The adhesion efficiency was assessed. At the same time the cell density of seeding cells was adjusted to 20×106/mL,10×106/ mL,and 5×106 /mL respectively, then the cells of different concentrations were seeded on the corner areas of the same DBM respectively. The growth and the distribution of human BMSCs on pDBM, were observed under fluorescence microscope on the 2nd, and 7th days following seeding. Cell-pDBM area ratio is assessed. Results The adhesion efficiency of Dil labeled and unlabeled BMSCs were (99.9±0.2)% and (99.1±1)% respectively, and no significant difference (P > 0.05) was found between two groups. There was no difference in cell- pDBM area ratio between different seeding concentrations. Conclusion Seeding of BMSCs on DBM at the concentration of 20×106/mL,10×106/mL,and 5×106/mL can all reach the purpose of covering the surface of DBM with cells.