Mouse 3t3-l1 Culture, Identification, And Berberine On The Proliferation And Differentiation

Part one Culture and evaluation of3T3-L1 preadipocyteObjective To culture mouse preadipocyte line 3T3-L1 and investigate the process in which 3T3-L1 preadipocyte proliferates and differentiates into mature adipocyte in the serum or serum-free medium, in order to study the effects of hormone and cytokine on proliferation and differentiation of preadipocyte.Methods3T3-L1 preadipocyte was cultured in the serum and serum-free medium. The growth status and process of proliferation and differentiation of 3T3-L1 preadipocyte were determined by MTT spectrophotometry. The growth curve was drawn.The change of morphology during the differentiation of preadipocyte was observed. Lipid content in cells were estimated by red Oil O staining. Cell differentiation was determined by the curve of time-OD value. And cells were photoed.Results3T3-L1 preadipocyte was cultured successfully. In microscope, the mophology of 3T3-L1 preadipocyte was similar to fibroblast, and couldn't stained by red Oil O.In the medium of 10 percent calf serum, 3T3-L1 preadipocyte could paste wall after 15 hours of culture. Logarithm proliferative phase began from the third day. The number of cell increased quickly. Proliferative platform phase was achieved in the ninth day.In the serum-free medium, Logarithm proliferative phase began from the fourth to eighth day. Proliferative platform phase began from the eighth day. The cell apoptosis occurred in the twelfth day and number of cell decreased.Lipid content in cells increased quickly after four days by red Oil O staining when differentiating medium was added. The global lipid droplet can be observed on the fourth day after cells were grown to confluence. The peak of differentiation was achieved till two weeks. And about 95 percent of preadipocyte differentiated into mature adipocyte with simple or multi-lipid droplet. ConclusionsThe 3T3-L1 preadipocyte line is cultured successfully. In the serum-free medium, the proliferation of preadipocyte is maintained and the differentiation from preadipocyte to mature adipocyte can be induced.