A technique is described for isolation and culture of adult human hepatocytes from surgical liver biopsies. The mean cell yield was 1.75 ± 107 cells per gm liver and viability averaged 80%. Hepatocytes were maintained in primary culture for about 10 days. Cell morphology and histochemical characteristics were similar to hepatocytes in vivo. Bile canaliculi were observed by electron microscopy. Intracellular albumin was demonstrated up to the 7th day of culture; albumin secretion rate was maximal (0.6 ± 0.33 /tg per hr per 106 cells) 5 days after plating. These studies demonstrate that adult human hepatocytes can be isolated from surgical biopsies with high yield, and differentiated function can be maintained for several days.