Effect Of Basic Firbroblast Growth Factor With Different Concentrations Upon Inducing HASCs To Differentiate Into Adipose Cells

ObjectiveTo study the effect of the exogenous Recombinant Human FGF-basic with different concentrations upon inducing Human ASCs to differentiate into adipose cells and to find the optimum concentration of exogenous rh-bFGF by experimental research.Methods1. hASCs were isolated, cultured, expanded by using enzyme digestion method from the liposuction aspirate for primary culture and observed changes in cell function and morphology. The molecular expression of hASCs was detected by using cellular immunofluorescence methods. Only cultured cells for passage3were used for adipogenic differentiation and Oil Red O staining to identify the differentiation.2. When cells were cultured for passage3, hASCs using adipogenic supplement were divided into experimental group and blank group:the experimental group of adipogenic supplement was divided into4groups by adding the exogenous rh-bFGF10ng/ml、20ng/ml and40ng/ml, the blank group of adipogenic supplement was cultured without exogenous rh-bFGF. The oil red O staining could qualitative analysis on the time of newly forming adipocyte cells.3. The experimental group of adipogenic supplement after adding the exogenous rh-bFGF10ng/ml、20ng/ml and40ng/ml12h,24h,48h, MTT colorimetric analysis were used to calculate the proliferation rate of hASCs differentiate into adipocytes.4. When experimental groups of adipogenic for14d, Western blot was used to detect the effects of rh-bFGF on the expression of lipid droplets surface protein CIDEC at different stages during the culture.Results1. The original cultured hASCs present similar morphology with fusiform and fibroblasts. This kind of cell presents CD105positive expression. With the effect of adipogenic differentiation medium. hASCs proved to be able to differentiate into mature adipocyte and the Oil Red O stain can be prove the differentiation. The experimental group could obviously shorten the period of inducing hASCs to differentiate into adioicytes, and promote the proliferation of adipocytes.2. The formation rate and the proliferation of adipocytes in the group adding40ng/ml rh-bFGF were superior to those in the experimental group else and blank group. The average time of the newly formed lipid droplets which adding40ng/ml rh-bFGF was (11.5±1.9) h.3. The average absorbance of cell proliferation which adding40ml b-FGF was0.52±0.10after adding10ng/ml、20ng/ml and40ng/ml rh-bFGF12h,24h,48h.4. The proliferative rate of adipocytes increases at different levels with adding10ng/ml、20ng/ml and40ng/ml rh-bFGF group,the CIDEC expression quantity of adding40ng/ml rh-bFGF group was also superior to that in the experimental group and blank group.ConclusionsAdding rh-bFGF in hASCs adipogenic supplement could promote the proliferation of adipocytes and dramatically accelerates the program of hASCs differentiate to adipocytes, and the optimum concentration of rh-bFGF was40ng/ml.