Abstract

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

Author(s):: LUO Sichang; XU Yi; Institute of Biothermal Technology, University of Shanghai for Science and Technology, Shanghai 200093, China

Keywords:: Keywords: kidney freeze-thaw method decellularization cryoprotective agent vascular network extracellular matrix

Abstract:: 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.