Spinner culture:a simple method for large scale culture of hepatocytes

Objective To explore a simple method for large scale culture of hepatocytes.Methods Suckling pig hepatocytes were isolated by modified two step in situ collagenase perfusion method described by Seglen.The suspension of hepatocytes was inoculated into spinner flasks precoated by 1% polylysine,then the flasks were stirred at lower speed in a 37℃ incubator.The viability,glucose syntheses,G 6 Pase activity,concentration of albumin and LDH in the supernatant were determined dynamically and the ultrastructure of the cultural hepatocytes was observed.Results The hepatocytes in spinner group showed enhanced viability,glucose synthesis capacity and G 6 Pase activity when compared to those in control group.Tight junctions and bile canaliculus like channels between cells within aggregates were observed under TEM.Conclusion The function of hepatocytes cultured by spinner method in an incubator was well maintained.This study provides a simple and better culture method for a large scale culture of hepatocytes and bioartificial liver.