Journal of Cell Science, Vol 10, 167-179, Copyright © 1972 by Company of Biologists

Submitted on July 2, 1971

Effect of Method of Cell Isolation on the Metabolic Activity of Isolated Rat Liver Cells

¹ Clayton Laboratories, Department of Medicine, and the Department of Physiological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, U.S.A.

Rat liver cells isolated with a tissue press, by tetraphenylboron (TPB) chelation, or by hyaluronidase and collagenase digestion were compared as to morphology, cell yield, and biosynthetic activity. The cells were intact by light microscopy; ultrastructural changes were present on electron-microscopic examination of all cell types except those prepared by a modified enzyme incubation method. TPB chelation gave the largest and enzyme techniques the smallest yield of cells. All cell types incorporated labelled amino acids into cellular protein; however, amino acid incorporation was greatest in cells isolated by the revised enzyme technique. Only enzyme and mechanical cells incorporated acetate into cellular lipid. Cofactor supplementation was not required in the modified enzyme cells. Acetate incorporation was more sensitive to preincubation than was amino acid incorporation. Calcium, which was required to prevent aggregation of enzyme cells, inhibited amino acid incorporation moderately and acetate incorporation completely in mechanical cells.