Non enzymatic liver cells isolation: citrate-perchlorate technique.
In the present study we describe a non-enzymatic technique for the isolation of rat hepatocytes by perfusion of liver through portal vein. The perfusion media consist of 1 mM sodium perchlorate, 5 mM sodium citrate, 10 mM glucose, 129 mM NaCl and 0.1% bovine-serum albumin at pH 7.4. After purification through diatrizoate gradient, electron microscopical studies revealed that most of purified hepatocytes were well preserved and presented a normal ultrastructure, thus correlating with previous biochemical results. The present method enables the recovery of metabolically and morphologically normal hepatocytes.
Camberos
M C
MC
CEDIE, Hospital de Pediatría Dr. Ricardo Gutiérrez, La Plata, Argentina.
Gomez Dumm
C L
CL
D'Alessandro
V
V
Zuazquita
A
A
Cresto
J C
JC
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient; D002951:Citrates; D003973:Diatrizoate; D008099:Liver / Q000648:ultrastructure*; D008297:Male; D010472:Perchlorates; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D000077559:Sodium Citrate; D017670:Sodium Compounds
D002951:Citrates; D010472:Perchlorates; D017670:Sodium Compounds; D003973:Diatrizoate; D000077559:Sodium Citrate; C031068:sodium perchlorate