Effects Of Insulin-like Growth Factor-1 On Proliferation Of Rat Nucleus Fibrosus Cells In Vitro

Objective To reseach the biologic specificity and degeneration of the anulus fibrosus(AF) cells by culturing AF cells and observing their continuous changes,meanwhile the methods and the cell models were provided for tissue engineering and gene therapy,to investigate the dose-dependent and time-dependent effect of insulinlike growth factor-1(IGF-1) on proliferation of anulus fibrosus (AF) cells in vitroMethods Tissue samples of AF were obtained from rats,Cells were isolated and cultured in HAMF12/DMEM medium supplemented with 10%fetal bovine serum(FBS),Cells were seeded in 96-well plates at 1×10~5cells/well,After synchroni zation.,medium was replaced by HAMF12/DMEM containing 1%or 10%FBS with IGF-1,Dose-dependent and time- dependent effect were examined by MTT assay,After staining with the haematine eosin and the toluidine blue,the immunohistochemical method was used to study the synthesis of typeⅠcollagen.Results The primary cells mostly appeared as flattened and spindle shape,with the pseudopodia being stretching out,The cytoplasm was profoundly blue after stained with the toluidine blue,The immunohistochemical test revealed that there existed the positive expression of typeⅠcollagen on the cells,In the presence of 1%FBS,the role of IGF-1 was less striking effect on cell proliferation,in 10%FBS,IGF-1 had statistically significant mitogenic effects,The dose-dependent effect of IGF-1 on cell proliferation was found within 100μg/L,The time-dependent effect showed a significant elevationConclusion IGF-1 was efficient to stimulate cell proliferation of rat NF cells in vitro in a dose-dependent and time-dependent manner,The experimental study provides a simple and effective method for the culture of anulus fibrosus cell in vitro,It also form a firm foundation for facilitating the furthur study of the manipulation and modulation of biologic function and biologic treatments of disc...