Preparation of whole-kidney acellular matrix using peristaltic pump perfusion and identification the composition of matrix

Abstract

Objective To make a whole-kidney acellular matrix scaffold through the method of peristaltic pump perfusion and to identify whether the kidney acellular matrix reserves extracellular matrix proteins or not. Methods The male Sprague-Dawley (SD) rats, 200~250 g. Intravenous cathers were injected into the renal artery to establish the channels for perfusion, followed by the perfusion of decellularization solution by using peristaltic pumps. After the decellularization, the acellular matrix was observed under microscope after hematoxylin and eosin(HE) staining and fluorescence microscope after DAPI staining. Immunohistochemistry was performed to identify the composition of kidney acellualr matrix. Results The result of HE staining and DAPI staining demonstrate the whole removal of cellular material in kidney acelluar matrix. Immunohistochemistry confirmed the preservation of the natural expression type of extracellular matrix proteins including collagen types I and IV, fibronectin and laminin. Conclusions Peristaltic pump perfusion is a successful method to make the kidney acellular matrix, and this scaffold retains the critical proteins of natural extracellular matrix. Therefore the kidney acellular matrix is an ideal natural scaffold for renal tissue engineering. Key words: Kidney Failure; Peristalsis; Extracellular Matrix