Abstract
Stromal vascular cells (SVC) from the perirenal adipose tissue of castrated Hanwoo beef cattle steers were isolated and cultured in medium containing different concentrations of calcium and retinoic acid. The cells were stimulated to differentiate in defined medium until lipid accumulation was apparent. The cell viability of adipose tissue SVC were not affected by the different concentrations of calcium and retinoic acid in the media. No observable cytotoxic effect was apparent among the cells treated with various levels of calcium and retinoic acid. Oil-red O staining of differentiated adipose tissue SVC shows that as the level of calcium and retinoic acid decreased, the degree of adipocyte differentiation increased. This shows that manipulating the extracellular level of calcium and retinoic acid in perirenal adipose tissue SVC can result in inhibition or stimulation of adipogenesis. RT-PCR and western blot analysis of adipogenic related genes and proteins clearly showed that the reduction in the level of calcium and retinoic acid in the media, stimulated adipogenesis. The expression level of PPARγ, C/EBPα, SREBP-1c and aP2 were significantly upregulated by the diminution of the calcium and retinoic acid. The expression of major calcium buffering protein calreticulin was also down regulated thus, diminishing its inhibitory effect on PPARγ. Retinoic acid related receptors RARα and CRABP-II were also down regulated upon the reduction of calcium and retinoic acid which may account for the decreased inhibitory effect of retinoic acid on adipocyte differentiation. These results suggest the possible mechanism of action for the increased adipocyte differentiation in Hanwoo beef cattle adipose stromal vascular cells upon decreasing the extracellular calcium and retinoic acid concentration. These findings may be applied in the future to further develop micronutrient manipulations to increase intramuscular fat accumulation in Hanwoo beef cattle.
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