Graduate Thesis Or Dissertation
 

Regulation of adipose stromal-vascular cell differentiation in culture

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  • Primary cultures of stromal-vascular (S-V) cells from adipose tissue were used to investigate the regulation of preadipocyte development. Differentiation of S-V cells was found to be under hormonal control. Insulin and glucocorticoids are essential for S-V cell differentiation in culture. S-V cells from both newborn and mature pig adipose tissue and sera from both ages were used to examine the effect of age on preadipocyte development. S-V cells from newborn pigs replicated faster and appeared more responsive to serum borne factors influencing S-V cell growth and development in culture. Serum source (newborn vs mature) did not affect differentiation of S-V cells from newborn or mature pig adipose tissue. When sera from fed or fasted pigs were used to culture newborn pig S-V cells, fasted pig sera stimulated greater differentiation and decreased cell replication as indicated by DNA content of rat S-V cell culture. Lean pig serum compared to obese pig serum, increased differentiation activity in culture of S-V cells an effect which may be influenced by sex. When sera from rat and pig were subjected to gel filtration fractionation on Sephacryl S-200 column, the elution profiles of both sera were similar. Rat serum contained six additional peaks (280 nm) not present in pig serum. Rat serum fraction two (apparent molecular size 67-150 kD) promoted greater differentiation of S-V cells than other rat serum fractions or pig serum fraction two. Fraction three (apparent molecular size 17-43 kD) of both sera inhibited differentiation and lipid filling in cultures of S-V cells but only rat fraction three promoted cell proliferation. Rat and pig S-V cells have different morphology when differentiated. Differentiated rat S-V cells appeared as individual cells when cultured in serum free or serum supplemented medium while differentiated pig S-V cells appeared as individual cells in serum free medium and as a tight cluster of cells in serum supplemented medium. Both cells responded differently to sera obtained from pigs of differing ages and development of rat S-V cells was influenced by anatomic site.
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