Hi all,
I am writing this question to inquire about the isolation of mouse primary hepatocytes. As I am a completely beginner, after isolating the cells, it turn out that the viability is 50% which is too low I think. I have to adjust my protocol again to increase the viability.
Apart from that, I would like to ask you if there are any methods to check the purity of the cells in order to confirm that the cells that I've got are hepatocytes (not any non-hepatocytes reside in the liver).
Thanks in advance for responses.
In order to achieve higher viability before seeding the hepatocytes you can use a Percoll gradient or just Percoll/Medium mixture (just check some published protocols). If you seed on collagen/fibronection coated plates non-hepatocytes should mostly "disappear" after 12-24 hours since they will not adhere.
@Hanno
Thank you very much for advice.
By the way, do you have any references discussing about the inability for adherence of non-hepatocytes? As far as I know, to isolate the kupffer cell, they also use collagen coated dish. So, I think Kupffer cell also adhere to the dish as hepatocytes.