存储条件对骨髓间充质干细胞的生物学特性和修复脊髓损伤效能的影响

The effect of storage conditions on the biological characteristics and repair potential of bone marrow mesenchymal stem cells after spinal cord injury

目的探讨0.9%生理盐水(Saline)、0.01M磷酸盐缓冲液(phosphate buffered saline,PBS)对骨髓间充质干细胞(bone marrow stem cells,BMSCs)存活、增殖等生物学特性以及对修复脊髓损伤(spinal cord injury,SCI)模型大鼠损伤效能的影响。方法体外,分离培养C57BL/6小鼠BMSCs,进行生理盐水、PBS及增殖培养基(PCM)处理不同时间,通过台盼蓝染色法检测各组细胞的活力,通过Brd U法检查各组细胞增殖情况。体内,通过LFB染色法检测各移植组SCI大鼠脊髓组织髓鞘再生情况;通过BBB运动功能评分检测大鼠运动功能恢复情况。结果存储液处理的BMSCs呈现出时间依赖的存活与增殖能力下降,PBS-BMSCs组存活率下降程度小于Saline-BMSCs组。移植组大鼠运动功能和损伤脊髓横断面髓鞘化程度显著恢复,并且PBS-BMSCs治疗组优于Saline-BMSCs治疗组。结论存储液处理BMSCs对SCI有治疗作用。

Objective To investigate the effect of saline and PBS used at different storage duration on the viability and proliferation of bone marrow stem cells （BMSCs） and repair potential after spinal cord injury（SCI）. Methods In vitro, BMSCs were isolated and cultured from C57BL/6 mice, and treated with saline, PBS and proliferation medium（PCM） for different time, the viability of BMSCs was detected by trypan blue staining, and the cell proliferation was detected by BrdU assay. In vivo, LFB staining method was used to detect the regeneration of myelin in spinal cord of SCI rats. The motor function was measured by BBB motor function score. Results PBS treated BMSCs showed decreased survival and proliferation capacity with a time-dependent manner, but mueh better than saline group. The area of myelin sheath in the spinal cord of the transplanted group was significantly recovered, with better in PBS group than in saline group. In addition, transplantation group can promote the recovery of motor function, with better in PBS-BMSCs than in saline-BMSCs group. Conclusion Storage solution treated BMSCs have therapeutic effect on SCI.