Establishment of a preadipocyte cell line from the epididymal fat pad of the lean C57 BL/6J mouse—Long term effects of insulin and triiodothyronine on adipose conversion

Abstract

A clonal cell line has been established from the epididymal fat pad of the C57 BL/6J +/? mouse. This line, designated HGFu, is aneuploid and exhibits both morphological and biochemical properties characteristic of mature adipocytes. Adipose conversion begins after confluence and is accompanied by (a) an early emergence of lipoprotein lipase, (b) an increase in the incorporation of [14C]acetate into lipids and in the activities of acid:CoA ligase and glycerol-3-phosphate dehydrogenase, (c) a 27- to 35-fold increase in the average triglyceride content per cell. In the presence of a beta-agonist (isoproterenol) a full lipolytic response (measured by fatty acid release) is observed with differentiated cells, whereas the responsiveness examined by cyclic AMP (cAMP) production is present both in undifferentiated and differentiated cells. Adipose conversion, estimated by activities of enzyme markers, is accelerated by the continuous presence in the culture medium of insulin and triiodothyronine both within their physiological range of concentrations, whereas insulin at supraphysiological concentrations shows a growth promoting activity. The concentrations of insulin and triiodothyronine required for half-maximal lipogenic effects are in agreement with the Kd values of their respective high affinity binding sites present in HGFu cells. The HGFu cell line seems to be a useful model for the study on a long term basis of the mechanisms of action both of insulin and triiodothyronine. Moreover it will make it possible to realize comparative studies between clonal lines established from the lean adult mouse (HGFu line) and from the genetically obese adult mouse (Ob17 line).