Published online Feb 15, 1998. doi: 10.3748/wjg.v4.i1.74
Revised: June 19, 1997
Accepted: August 3, 1997
Published online: February 15, 1998
AIM: To explore a simplified method for isolation of hepatocytes and establish a method of primary hepatocyte culture with more aggregates and longer persisence.
METHODS: Wistar rat hepatocytes were isolated by a single extracorporeal two-step perfusion method, and the cells were seeded on poly-HEMA coated flasks and cultured with hormonally defined medium and gentle shaking at regular intervals.
RESULTS: The total yield of isolating hepatocytes amounted to 108 cells for each rat liver with the viability of more than 90% in all isolations. Under the nonadherent environments, the cells were found to attach to each other and form multicellular aggregates rapidly, and the aggregates became spheroidal shape after two days in culture. The morphologic characteristics and albumin synthetic function of the multicellular spheroidal aggregates can be maintained for one month.
CONCLUSION: The simple and reliable isolation as well as large scale and longer time culture of hepatocytes can be used for experiments in liver cell transplantation and bioartificial liver support system.