Adhesive culturing of hepatotic microcarriers and function measurement in rat

Objective To study a simple culturing method for obtaining hepatocytes at high cell density and high activity.Methods Adult Sprague Dewley rats'( n =5) hepatocytes were harvested and isolated by a modified enzymatic isolation technique, called semi situ collagenase digestion technique. The harvested hepatocytes were cultured in a conditioned medium on microcarriers cytodex 3 with the restriction attachment , regularly twirled and growth factors were added into it. Parallel cultures of plated hepatocytes were also conducted. Morphology of the hepatocytes was examined dynamically. The functions of albumin secretion and glucose syntheses in hepatocytes were measured.Results The cultured lymphocytes on microcarriers had a higher survival rate, function of albumin secretion and glucose syntheses, which are superior to those of plated hepatocytes.Conclusions The microcarrier's culture method can provide more liver cells, which have a higher viability and better function, for hepatocyte transplantation, liver disease therapy and bioartificial liver.