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Hormonal regulation of postnatal chicken preadipocyte differentiation in vitro.

Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology (2003-10-08)
T G Ramsay, R W Rosebrough
ABSTRACT

This study was designed to develop a culture system from the stromal-vascular fraction of chicken adipose tissue that can be used to characterize hormones that promote preadipocyte differentiation. Abdominal adipose tissue was excised from 2 to 4-week-old male broilers (Gallus domesticus) by sterile dissection. The stromal-vascular cell fraction from the adipose tissue was isolated by collagenase digestion, filtration, and subsequent centrifugation. These preadipocytes were seeded in six well culture plates and proliferated to confluency in 10% fetal bovine serum in DMEM/F12 (50:50) medium. At confluency, experiments were initiated to determine hormonal requirements for differentiation. Insulin (100 nM) stimulated expression of citrate lyase and sn-glycerol-3-phosphate dehydrogenase relative to lactate dehydrogenase in the presence of 2.5% chicken serum (P<0.05), but not with 10% chicken serum (P>0.05). Triiodothyronine (T(3), 1 nM) and insulin-like growth factor 1 (100 ng/ml) had no effect on differentiation. Dexamethasone (Dex, 1 microM) stimulated differentiation in 2.5 or 10% chicken serum (P<0.05). Insulin, Dex and 2.5% chicken serum stimulated enzymatic differentiation to the extent of 10% chicken serum, but heparin (10 U/ml) addition, in combination with insulin and Dex was necessary to stimulate lipid filling of adipocytes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Citrate Lyase from Klebsiella pneumoniae, lyophilized powder, ≥0.20 unit/mg solid
Sigma-Aldrich
Insulin from porcine pancreas, powder, ≥27 USP units/mg