生物蛋白胶作为骨髓间充质干细胞移植载体的体外相容性研究

An in vitro study on biocompatibility of bioprotein glue used as carrier for BMSCs transplantation

[目的]构建骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)-生物蛋白胶复合体评价其生物相容性,探讨其作为种子细胞载体的可行性。[方法]本实验分为两组,实验组5×107骨髓间充质干细胞(BMSCs)与生物蛋白胶相混制备MSC-生物蛋白胶复合体,空白组骨髓间充质干细胞(BMSCs)。通过Dead/live染色观察两组的细胞形态及21 d内细胞的存活情况,同时分别在1、3、7、14 d各取上清进行Elisa检测并计算其分泌释放NGF和BDNF的浓度。[结果]实验组培养3 d后光镜下观察呈多个突起,细胞形态良好,空白组呈菱形和多角形;Dead/live染色结果提示体外培养1 d,两组细胞的存活率都在90%,比较无差异,无统计学意义(P﹥0.05),7、14、21 d两组细胞的存活率有差异,具统计学意义(P﹤0.05),存活率在60%以上;Elisa检测结果显示两组中NGF在1 d时有差异,具统计学意义(P﹤0.05),3、7、14 d时NGF和BDNF的含量比较无统计学意义(P﹥0.05),结果显示生物蛋白胶对MSC分泌的细胞因子NGF和BDNF的释放没有明显影响。[结论]生物蛋白胶是一种理想的可用于临床的细胞载体,具有良好的生物相容性,BMSCs-生物蛋白胶复合体能持续稳定的释放神经生长因子BDNF和NGF。

[ Objective] To construct the complex of bone marrow mesenchymal stem cells （BMSCs） and fibrin glue, and evaluate its biocompatibility and feasibility as a seed cells carrier. [ Methods ] Experimental group was established with the MSC - fibrin glue complex of 5 x 107 bone marrow mesenchymal stem cells （BMSCs） and fibrin glue, the control group with BM- SCs only. The cell morphology and viability were observed at days 21 by Dead / hve staining. The concentrations of released NGF and BDNF were determined by Elisa at 1, 3, 7, 14 days post -seeding, respectively. [ Results] Good cell morphology was observed in experimental group cultured for 3 days by the light microscope, control group showed diamond - shaped and po- lygonal. Dead / live staining results indicated that there was no statistical significance in survival rate between the two groups cul- tured for 1 day in vitro （90%） （P 〉 0. 05）, however at days 7, 14, 21 there was statistical significance （ P 〈 0. 05 ）, all of the survival rate was above 60%. Elisa test results indicated that there was statistically significant difference on release of NGF between the two groups at day 1 （ P 〈 0. 05 ）, however there was no statistical significance at days 3, 7, 14 （ P 〉 0.05 ） . [ Conclusion] BMSCs -fibrin glue complex has good biocompatibility and releases BDNF and NGF continually. It is a promising cell carrier.