Rong Fu Liu1, Jia sheng Gao2, Yu Feng Yang3, Wei Xin Zeng3. 1. Department of Urology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China. lliurf@126.com. 2. Department of Urology, the First People Hospital of Jiujiang City, Jiujiang 332000, China. 3. Department of Urology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
Abstract
PURPOSE: To design a whole-kidney a cellular matrix scaffold using peristaltic pump perfusion and to ascertain the retention of extra cellular proteins by the scaffold. MATERIALS AND METHODS: Male Sprague-Dawley (SD) rats weighing 200-250 g were used. Intravenous catheters were inserted into the renal artery followed by perfusion of decellularization solution using a peristaltic pump. After decellularization, the acellular matrix was observed under a microscope after hematoxylin and eosin (H&E) staining and a fluorescence microscope after 4′,6-diamidino-2-phenylindole (DAPI) staining. Immunohistochemistry was used to identify the composition of kidney acellular matrix. RESULTS: The result of H&E and DAPI staining demonstrate the removal of cellular material in kidney a cellular matrix. Immunohistochemistry confirmed the conservation of the natural expression of extra cellular matrix proteins including collagen types I and IV, fibrin and laminin. CONCLUSION: Peristaltic pump perfusion enables successful preparation of renal a cellular matrix, to retainthe criticalproteins of natural extra cellular matrix. The resulting kidney a cellular matrix represents an ideal natural scaffold for renal tissue engineering.
PURPOSE: To design a whole-kidney a cellular matrix scaffold using peristaltic pump perfusion and to ascertain the retention of extra cellular proteins by the scaffold. MATERIALS AND METHODS: Male Sprague-Dawley (SD) rats weighing 200-250 g were used. Intravenous catheters were inserted into the renal artery followed by perfusion of decellularization solution using a peristaltic pump. After decellularization, the acellular matrix was observed under a microscope after hematoxylin and eosin (H&E) staining and a fluorescence microscope after 4′,6-diamidino-2-phenylindole (DAPI) staining. Immunohistochemistry was used to identify the composition of kidney acellular matrix. RESULTS: The result of H&E and DAPI staining demonstrate the removal of cellular material in kidney a cellular matrix. Immunohistochemistry confirmed the conservation of the natural expression of extra cellular matrix proteins including collagen types I and IV, fibrin and laminin. CONCLUSION: Peristaltic pump perfusion enables successful preparation of renal a cellular matrix, to retainthe criticalproteins of natural extra cellular matrix. The resulting kidney a cellular matrix represents an ideal natural scaffold for renal tissue engineering.