mouse liver perfusion and hepatocyte isolation - Is perfusion absolutely necessary? (Jun/20/2006 )
I have a question about techniques used to isolate fresh mouse heptocytes. I am pretty sure that perfusion methods are used to optomize the recovery of viable cells. However, I was wondering if it's possible to isolate liver cells from a freshly excised liver that is quickly minced and soaked in collagenase? I realize that this may be a niave question, so I apologize in advance for my ignorance.
I read some papers in which some authors did a partial perfused digestion of the liver and after they continued the digestion in vitro. But in my opinion is very diffucult to recovery viable cells w/o liver perfusion, in fact also when human hepatocytes are isolated (normally you have only pieces of liver), the authors prefer to pefusing the liver, of course this is very difficult in case you have only small pieces and in this case authors did a in vitro perfusion. Hovewer me too I tried but I didn't be able to recovery no viable cells. The best choice is to have mice and perfuse after deep anesthesia. Be carefull with the mechanical stress beacuase hepatocytes are normally very fragile cells. Good luck
Thanks for your reply. Do you mind listing one or two of the papers that describe partial perfusion?
i suggest you to read these paper:
1) Wu W, Kocarek TA, and Runge-Morris M (2001) Sex-dependent regulation by dexamethasone of murine hydroxysteroid sulfotransferase gene expression. Toxicol Lett 119: 235-246.
2) Pichard L, Raulet E, Fabre G, Ferrini JB, Ourlin JC, Maurel P. Human hepatocyte culture. Methods Mol Biol. 2006;320:283-93.
Good luck again
Nobody know if there are counter-indications to perfuse the liver trough the heart in term of viablity of the cells? Notably in mice?