[Comparison of bidirectional collagenase perfusion with one way perfusion in the isolation of periportal or perivenous hepatocytes in rats].
To selectively isolate hepatocytes from the periportal (PP) and perivenous (PV) regions of rat liver acinus, we compared two different perfusion methods with collagenase: the bidirectional perfusion method (2-P) and the one way perfusion method (1-P). We determined the optimal conditions for each method on the basis of the zonal selectivity of isolated hepatocytes with a hematoxylin-eosin stained liver specimen. By both methods, hepatocytes were selectively isolated from the PP and PV regions. Comparing cell yield and cell viability after the two perfusion methods, 1-P was found to be better than 2-P. Density gradient centrifugation with Percoll was found to be an effective procedure for removing the damaged hepatocytes. We concluded that 1-P could isolate viable PP and PV hepatocytes with a normal glucagon-cyclic AMP response and ultrafine structure in high yield from rat liver.
Yamazaki
H
H
Department of Pharmacology, Faculty of Medicine, Toyama Medical and Pharmaceutical University, Japan.
Nishiguchi
K
K
Nakanishi
S
S
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D008099:Liver / Q000166:cytology*; D008297:Male; D003012:Microbial Collagenase / Q000008:administration & dosage*; D010477:Perfusion; D051381:Rats; D011919:Rats, Inbred Strains
D003012:Microbial Collagenase