Culturing and assays function of rat hepatocytes on microcarriers

Objective:To study the simplified method of isolation and culture of rats' hepatocytes at high cell density and high activity.Method Adult Sprague Dawley rats hepatoeytes(n=5)were harvested by a modi- fied enzymatic isolation technique,called semi-situ collagenase digeston technique.Harvested hepatocytes were cultured in a conditioned medium on microcarriers cytodex-3 with the restriction attachment,regular twirled and adding growth factors.Parallel cultures of plated hepatocytes were also conducted.Morphology was observed dynamically.Functions were evaluated by albumin syntheses and glucose syntheses.Results: The viabltiy of aggregate hepatocytes with preservation of normal morphologic was higher than monolayer hepatocytes(67.3±5.3％,38.5±6.6％,p0.05)in the 3rd weeks.The microcarrier cultured hepatocytes showed significant higher acitivity in albumin secretion compared with monolayers(p0.05).Significant higher glucose syntheses were also noted in micraoearrier cultured hepatocytes than plated hepatocytes in the first day and 7th day culture(p0.05). Conclusion:The microcarriers culture method can provide enough more liver cells,which were have higher viability,greater function,for hepatoctytes transplantation,liver disease therapy and bioartificial liver.