mouse hepatocytes isolation

[amicia]

Thank Admin, I'm in a bad situation So all suggestions are welcomed

[IoDiNE]

any news on this front?

[amicia]

No I'm sorry, I'm going on to perfuse trough portal vein but w/o any appreciable results. So I'm thinking to move into a lab in which routinely is done

[admin]

Hello Amicia!

the group that does hepatocyte isolations basically told me that they are always very carefull to:

1) not to lose too much blood
2) not to damage vessel when perfusing
3) when perfusing use a pump
4) make sure you do not injure the animal (organs, vessels) before you do perfusion
5) try to prevent hemmorraging of the blood vessel when perfusing

I will try to get back to you with more tips

[amicia]

Hello Admin,
thank you very much for the help. I'm in agreement with them, but I didn't understand the point 1 and the point 5. Of course when the perfusion starts you lose the blood. I thought the only think was to avoid the cloatting of the blood

[amicia]

hi All,
just to tell you I resolved my problems, was a bad batch of collagenase that killed alls my cells. thank you very much for your assistance. A special thank to Admin, Kiki06 and Moleculardude.

[bioinfofreak]

wow! it pays to check your buffers! what was that buffer control thread I just posted too! that guy may be onto something!

Good Work!