The first technique for large-scale preparation of isolated hepatocytes was described in 1953 and involved perfusion of rat liver under pressure with a Ca(2+)-free solution containing a chelating agent. Various modifications of this technique were in use over the next ten years, until it was demonstrated that cells prepared in this manner were grossly damaged, losing most of their cytoplasmic enzymes during the preparative procedure. The successful preparation of intact isolated hepatocytes by collagenase-treatment of liver was achieved in 1967, and the widespread use of intact hepatocyte suspensions was accelerated by the development soon after of high-yield preparative techniques involving perfusion of the liver with a medium containing collagenase. The introduction of the isolated hepatocyte preparation has enabled experimental studies that otherwise would not be feasible. Important advances have been the use of cultured hepatocytes, frequently of human origin, for the investigation of the metabolism and toxicology of potential therapeutic agents. Success in this field has been achieved through the steady improvement in techniques for the maintenance in culture of differentiated hepatocytes, and in particular their cytochrome P450 complexes. Another area showing considerable promise is the employment of hepatocytes, generally from a porcine source, in temporary support systems for patients with acute liver failure. Our own studies have concentrated on the demonstration of long-range interactions between hepatocyte compartments which suggest that energy transfer between cell compartments can take place without ATP turnover.
The sphincter of Oddi (SO) regulates trans-sphincteric flow (TSF) by acting primarily as a pump or as a resistor in specific species. We used the Australian possum SO, which functions similarly to the human SO, to characterize SO motility responses to different common bile duct (CBD) and duodenal pressures. Possum CBD, SO and attached duodenum (n =18) was mounted in an organ bath. External reservoirs were used to impose CBD (0-17 mmHg) and duodenal (0, 4, 7 mmHg) pressure. Spontaneous SO activity was recorded using four-lumen pico-manometry and TSF was measured gravimetrically. Temporal analysis of manometric and TSF recordings identified three functionally distinct biliary-SO regions, the proximal-SO (juxta-CBD), body-SO and papilla-SO. At CBD pressures < 3 mmHg the motor activity of these regions was coordinated to pump fluid. Proximal-SO contractions isolated fluid within the body-SO. Peristaltic contraction through the body-SO pumped this fluid through the papilla-SO (17-27 µl contraction), which opened to facilitate flow. CBD pressure > 3.5 mmHg resulted in progressive changes in TSF to predominantly passive 'resistor'-type flow, occurring during proximal-SO-body-SO quiescence, when CBD pressure exceeded the pressure at the papilla-SO. Progression from pump to resistor function commenced when CBD pressure was 2-4 mmHg greater than duodenal pressure. These results imply that TSF is dependent on the CBD-duodenal pressure difference. The papilla-SO is pivotal to TSF, relaxing during proximal-SO-body-SO pumping and closing during proximal-SO-body-SO quiescence. The pump function promotes TSF at low CBD pressure and prevents bile stasis. At higher CBD pressure, the papilla-SO permits TSF along a pressure gradient, thereby maintaining a low pressure within the biliary tract.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.