Development of a bioassay system for investigating insulin resistance factors of pregnancy
To determine if late-term pregnant serum and/or placenta could induce insulin resistance in normal adipose cells, the authors have developed an insulin sensitive bioassay system. Cells isolated from epididymal fat pads of 250-275 g Sprague Dawley rats are preincubated for 3 hours at 37/sup 0/ in media 199 and serum or placental extract. The cells are washed free of serum and tested for metabolic activity in a 2 hour incubation which measures the conversion of U-/sup 14/C-glucose to /sup 14/CO/sub 2/ and to /sup 14/C-triglyceride fatty acids under basal and insulin stimulated conditions. Maximal insulin responsiveness (350-450% basal for CO/sub 2/ and 1400-1700% basal for fatty acids) is achieved using Worthington Type II collagenase and a 45-60 minute digestion period for cell isolations and Krebs-Ringer bicarbonate buffer containing 0.5 mM glucose, 2% Armour bovine serum albumin (CRG-7), 1000 ..mu..U/ml insulin and 110,000 to 120,000 cells in the 2 hour incubations. Using this bioasssay system the authors have found that insulin responsiveness, in terms of glucose conversion to fatty acids, is unchanged when cells are preincubated with 5% control pig serum but reduced following preincubation with late pregnant (110 day) pig serum. In future experiments the authors hope to further characterize the factor(s) in pregnant serum responsible for inducing this metabolic effect.
- Research Organization:
- Univ. of Georgia, Athens
- OSTI ID:
- 5508930
- Report Number(s):
- CONF-8604222-; TRN: 86-026561
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
INSULIN
BIOASSAY
METABOLISM
CARBON 14 COMPOUNDS
CARBON DIOXIDE
CARBOXYLIC ACIDS
FAT CELLS
GLUCOSE
PLACENTA
PREGNANCY
RATS
TRACER TECHNIQUES
ALDEHYDES
ANIMAL CELLS
ANIMALS
CARBOHYDRATES
CARBON COMPOUNDS
CARBON OXIDES
CHALCOGENIDES
CONNECTIVE TISSUE CELLS
FETAL MEMBRANES
HEXOSES
HORMONES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MEMBRANES
MONOSACCHARIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXIDES
OXYGEN COMPOUNDS
PEPTIDE HORMONES
RODENTS
SACCHARIDES
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques