Long-term culture of primary rat hepatocytes with high albumin secretion using membrane-supported collagen sandwich.

Primary cultured rat hepatocytes in a membrane-supported collagen sandwich maintained their normal cell morphology and high level of albumin secretion for over 56 days. It was found that the existence of an upper layer of collagen gel is crucial for long-term culture and that the transference of cellular nutrients between the culture media and hepatocytes from both the upper and the lower sides of gel layers promotes albumin secretion. These facts suggest that the membrane-supported collagen sandwich mimics well the in vivo environment of hepatocytes. This method has great potential for the long-term culture of primary cells.