北京生物医学工程

海藻糖对大鼠肾脏冻融法脱细胞效果的影响

Effect of trehalose on decellularization of rat kidney by freeze-thaw method

作者：罗嗣昌胥义党航宇柳珂

单位：上海理工大学生物系统热科学研究所（上海200093）

分类号：R318.04

出版年·卷·期（页码）：2020·39·3（227-232）

摘要：

目的应用海藻糖溶液辅助冻融法脱细胞获得脱细胞支架,优化大鼠肾脏冻融脱细胞工艺?方法首先采用不同浓度的海藻糖溶液对大鼠肾脏进行预处理,放入-20℃环境下进行冷冻,12h后在37℃的温水中进行水浴复温;然后采用TritonX?100以0mL/min流速对肾脏24h灌注脱细胞;最后通过血管网络CT三维重构?染色切片?蛋白质定量检测以及力学性能检测等手段评估所得大鼠脱细胞肾脏支架,同时与新鲜组做对比?结果未添加海藻糖实验组血管网络损伤较大,洗脱效果一般,无法制取有效的脱细胞支架?添加不同浓度海藻糖实验组的血管网络也均有一定的损伤?但5%海藻糖组保留最为完整,能够为脱细胞支架提供良好的血管网络,细胞外基质保留完整且洗脱细胞效果良好?结论在一定浓度下海藻糖能够更好地辅助冻融法脱细胞获得更优大鼠肾脏脱细胞支架,这为下一步研究冻融脱细胞获取脱细胞肾脏支架提供了重要的实验依据?

Objective Trehalose is used to pretreat rat kidney for obtaining acellular scaffold with freeze-thaw method, and the freeze-thaw acellular process of rat kidney is optimized. Methods.The samples were pretreated with concentrations of trehalose and frozen at-20 ℃ for 12 hours. Then the kidneys were reheated in 37 ℃ warm water, and they were perfused with TritonX-100 at the flow rate of 0.4mL/min for 24 hours. Some assessment methods including CT three-dimensional reconstruction, staining section, protein quantitative detection and mechanical properties detection and so on, were utilized to investigate the integrity of vascular structure inside the acellular renal scaffolds and other functional factors, and compared with the fresh group. Results The vascular network of the samples without trehalose was damaged significantly, and decellularization effect was general, and the effective acellular scaffold could not be made. To some extent, the vascular network was damaged in the experimental group with trehalose at different concentrations.The 5% trehalose group was the best one, which could provide a relatively integral vascular network and other functional factors for acellular scaffold. Conclusions Comparing the results of different experimental groups, we can know that trehalose can better assist the freeze-thaw acellular method to obtain better acellular scaffolds in rat kidney,this results provide an important experimental basis for the future study of freeze-thaw acellular renal scaffolds.

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