Preparation of parenchymal and non-parenchymal cells from adult human liver--morphological and biochemical characteristics.
By perfusion of the isolated human liver with collagenase and hyaluronidase a mixed suspension of various cell types was obtained. Pure parenchymal cells were prepared by differential centrifugation, pure non-parenchymal cells by the use of pronase and subsequent isopycnic centrifugation on metrizamide gradients (50-300 g/l). About 90% of the parenchymal and non-parenchymal cells were viable as judged by trypan blue staining. Non-parenchymal cells were not capable fo gluconeogenesis but at high rates. Parenchymal cells retained their ability to form glucose and to accumulate glycogen from fructose greater than lactate/pyruvate greater than alanine. Studies on binding of 125I-labelled insulin by isolated parenchymal cells were performed at 30 degrees C. The binding data may fit with a minimum of two classes of binding sites: (a) high affinity--low capacity sties (Kd approximately 6.6 nmol/l, capacity approximately 16 000 insulin molecules per cell) and (b) low affinity-high capacity sites (Kd approsimately 0.37 mumol/l, capacity approximately 646 000 molecules per  cell).
D000328:Adult; D002469:Cell Separation / Q000379:methods; D005943:Gluconeogenesis; D006801:Humans; D006867:Hydrolases; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008112:Liver Glycogen / Q000096:biosynthesis; D011972:Receptor, Insulin / Q000032:analysis; D014461:Ultracentrifugation / Q000379:methods
D008112:Liver Glycogen; D011972:Receptor, Insulin; D006867:Hydrolases