Preparation of Parenchymal and Non-Parenchymal Cells from Adult Human Liver — Morphological and Biochemical Characteristics

Publisher Website
Google Scholar
Abstract
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 of gluconeogenesis but utilized glucose at high rates. Parenchymal cells retained their ability to form glucose and to accumulate glycogen from fructose > lactate/pyruvate > alanine. Studies on binding of 125I-labeled insulin by isolated parenchymal cells were performed at 30.degree. C. The binding data may fit a model with a minimum of 2 classes of binding sites: high affinity - low capacity sites (Kd .apprx. 6.6 nmol/l, capacity .apprx. 16,000 insulin molecules/cell) and low affinity - high capacity sites (Kd .apprx. 0.37 .mu.mol/l, capacity .apprx. 646,000 molecules/cell).