Procedure for Decellularization of Rat Livers in an Oscillating-pressure Perfusion Device.
Decellularization and recellularization of parenchymal organs may enable the generation of functional organs in vitro, and several protocols for rodent liver decellularization have already been published. We aimed to improve the decellularization process by construction of a proprietary perfusion device enabling selective perfusion via the portal vein and/or the hepatic artery. Furthermore, we sought to perform perfusion under oscillating surrounding pressure conditions to improve the homogeneity of decellularization. The homogeneity of perfusion decellularization has been an underestimated factor to date. During decellularization, areas within the organ that are poorly perfused may still contain cells, whereas the extracellular matrix (ECM) in well-perfused areas may already be affected by alkaline detergents. Oscillating pressure changes can mimic the intraabdominal pressure changes that occur during respiration to optimize microperfusion inside the liver. In the study presented here, decellularized rat liver matrices were analyzed by histological staining, DNA content analysis and corrosion casting. Perfusion via the hepatic artery showed more homogenous results than portal venous perfusion did. The application of oscillating pressure conditions improved the effectiveness of perfusion decellularization. Livers perfused via the hepatic artery and under oscillating pressure conditions showed the best results. The presented techniques for liver harvesting, cannulation and perfusion using our proprietary device enable sophisticated perfusion set-ups to improve decellularization and recellularization experiments in rat livers.
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Karl
K
Hillebrandt
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Dietrich
D
Polenz
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Antje
A
Butter
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Peter
P
Tang
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Anja
A
Reutzel-Selke
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Andreas
A
Andreou
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Hendrik
H
Napierala
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Nathanael
N
Raschzok
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Johann
J
Pratschke
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin.
Igor M
IM
Sauer
General, Visceral, and Transplantation Surgery, Charité - Universitätsmedizin Berlin; benjamin.struecker@charite.de.
Benjamin
B
Struecker
D000818:Animals; D003584:Cytological Techniques / Q000295:instrumentation* / Q000379:methods*; D005109:Extracellular Matrix; D008099:Liver / Q000166:cytology*; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods