Increased Adipogenesis of Human Adipose-Derived Stem Cells on Polycaprolactone Fiber Matrices
Insulin stimulated glucose transport was measured as uptake of deoxy-D-glucose 2-[1- 14C] in response to indicated doses of insulin during 60 min and normalized between 0 and 100% glucose uptake and outliers were excluded using the tukey test in human preadipocytes differentiated in 2D (solid black line, black dots), n = 19 differentiated at three different occations human preadipocytes differentiated on aligned matix (green line and dots), n = 18 differentiated at three different occations, human preadipocytes differentiated on random matrix (magenta line and dots), n = 17 differentiated at three different occations, human mature adipocytes (dashed black line, open circles) measured as a mean of duplicates isolated from 7 different subjects. A) All four data sets normalized to min 0 and max 100% and fitted to the same slope, inserted: EC50 values from the dose-responses, statistical testing was performed using students t-test, * denotes a p-value lower than 0.05. B) Glucose uptake, basally and stimulated with 100 nM insulin, normalized to total protein content. C) All four data sets normalized to min 0 and max 100% and fitted to individual, three parameter slopes. D) Insulin receptor levels measured as arbitrary units and normalized to actin content with western blot. Differentiated human preadipocytes(n = 6, N = 3) or human mature adipocytes N = 7 E) Fully differentiated human preadipocytes N = 3 or human mature adipocytes, N = 6, were stimulated with indicated concentrations of insulin for 10 minutes subjected to SDS-PAGE and western blot, phosphorlylation of serine 473 of PKB measured as arbitrary units and normalized to actin content and maximal phosphorylation was set to 100%.