不同保护剂对大鼠肾脏冻融法脱细胞的影响

目的:利用不同保护剂对大鼠肾脏进行预处理，辅助冻融法脱细胞获得脱细胞支架，优化冻融脱细胞工艺。方法:采用不同保护剂预处理大鼠肾脏，放入-20℃环境下进行冷冻，12 h后进行37℃水浴复温，然后利用Triton X-100灌注24 h、PBS灌注2 h。最后通过CT三维重构、染色切片、蛋白质定量以及力学性能分析等手段评估所得脱细胞支架。结果:未添加保护剂组血管网络损伤较大，洗脱效果一般。添加不同保护剂组的血管网络也均存在一定的损伤，但其中10%DMSO和5%海藻糖组血管网络保留得较为完整。但10%DMSO组DNA等物质残留过多，5%海藻糖组洗脱细胞效果良好。结论:加载保护剂能够对冻融法脱细胞起到一定的促进作用，且能保护大鼠肾脏内部血管网络等结构，但不同保护剂对冻融法脱细胞的作用效果不同。

Effects of different cryoprotective agents on decellularization of rat kidney by freeze-thaw method

Abstract: Objective To obtain decellularization scaffold by freeze-thaw method with the assist of different cryoprotective agents (CPA) for preprocessing the rat kidney, and to optimize the decellularization process of freeze-thaw method. Methods The rat kidneys were preprocessed with different CPA and then frozen at -20 ℃. After 12 hours, the kidneys were reheated in 37 ℃ warm water, and then perfused with TritonX-100 for 24 hours and phosphate buffered saline for 2 hours. Finally, the obtained decellularization scaffolds were evaluated by CT three-dimensional reconstruction, staining sections, protein quantification and dynamic mechanical analysis. Results The vascular network of the sample without being pretreated with CPA was damaged significantly, and the decellularization effect was general. After that rat kidneys were preprocessed with CPA, the vascular networks were also damaged, but the vascular networks in 10%DMSO and 5%trehalose groups remained relatively intact. There were too many residues such as DNA in 10%DMSO group, and the decellularization effect of 5%trehalose group was better. Conclusion CPA can promote freeze-thaw decellularization and protect the internal vascular networks of rat kidneys, but different CPA has different effects on freeze-thaw decellularization.