[Preparation of whole-kidney acellular matrix in rats by perfusion].
OBJECTIVE
To prepare rat whole-kidney acellular matrix (ACM) scaffolds using fluid perfusion method.


METHODS
The kidneys with ureters and renal vessels were harvested from 12-week-old Wistar rats. Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney retrograde perfusion successively with heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 100 cmH2O. After decellularization, the scaffolds were observed under microscope with HE staining, scanning electron microscope, and fluorescence microscope with DAPI fluorescence staining.


RESULTS
No cell residue was found in the scaffolds under microscope. Scanning electron microscope identified reticular structures consisting of basilar membrane and collagen without normal cellular structures in the scaffolds, and no strong fluorescence due to the binding of DAPI to the cell nuclei was observed under fluorescence microscope.


CONCLUSIONS
Fluid perfusion is simple and reliable to prepare rat whole-kidney acellular matrix, which may serve as an ideal cell-free scaffold.
Liu
Chun-xiao
CX
Department of Urological Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. Liuchx888@hotmail.com
Liu
Si-ran
SR
Xu
A-bai
AB
Kang
Yu-zhan
YZ
Zheng
Shao-bo
SB
Li
Hu-lin
HL
D000818:Animals; D001672:Biocompatible Materials; D002469:Cell Separation / Q000379:methods*; D005109:Extracellular Matrix; D005260:Female; D007668:Kidney / Q000166:cytology*; D008297:Male; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
D001672:Biocompatible Materials