关键词: 去细胞, 肾, 细胞外基质, 血管铸型, 组织工程, 免疫荧光, 大鼠

Abstract: Objective To prepare a whole kidney decellularized extracellular matrix bio-derived scaffold and to perform preliminary identification. Methods The kidney with ureters and renal vessels was harvested from adult SD rats.Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney perfusion successively with heparinized PBS,0.05% trypsin,1%Sodium dodecyl sulfate（SDS）,1% Triton X-100 and antibiotic-containing PBS under a pressure of 3.6 mmHg in 37℃.After decellularization,the scaffold and native kidney were observed through genomic DNA content analysis,transmission electron microscopy,HE staining,immunofluorescence and vascular cast. Results Quantitative analysis of DNA content within the scaffold showed a 97% decrease compared to the native kidney.The agarose gel electrophoresis showed no DNA bands associated with the decellularized scaffold.Transmission electron microscope,HE and immunohistochemistry showed a lot of collagen fibers but no visible cell nuclei remained after decellularization.Cast specimen showed that renal arteries were more sparse,but still full and clear compared with the native kidney. Conclusion The method of combined enzymatic digestion and detergent washing,soaking through the infusion is a simple,ideal preparation for the bioengineering scaffold of the kidney,which enables the quick and consistent removal of all the cellular components of the tissue,leaving behind mostly intact the extracellular

Key words: Decellularization, Kidney, Extracellular matrix, Vascular cast, Tissue engineering, Immunofluorescence, Rat