Adipocyte formation is included in our quality control testing for the 3T3-L1 cell line (ATCC® CL-173) and all lots show clear indication of adipocyte differentiation. Our protocol includes allowing the cells to reach confluence and then observing for the formation of lipid droplets over approximately 2 weeks. We have not attempted to induce differentiation of 3T3-L1 with a cocktail mixture of dexamethasone, IBMX, and insulin.
In order to assure our customers that all lots of 3T3-L1 cells show adipocyte differentiation the following protocol is part of the QC process. At the ATCC we allow one flask to become confluent (using ATCC DMEM 30-2002 supplemented with ATCC calf serum 30-2030). The fluid is changed twice a week and the cells are observed for evidence of adipocyte formation. In almost all lots of 3T3-L1 cells fat accumulation begins about 5 days after 100% confluence and there is 50% differentiation to adipocytes after 15-16 days post confluence. Each and every field examined has foci of cells with cytoplasmic droplets consistent with those shown in Dr. Howard Green's publications.
We recommend that customers consult the following references concerning differentiation of 3T3-L1 cells:
Green H and Meuth M. An established pre-adipose cell line and its differentiation in culture. Cell 3: 127-133, 1974 PubMed: 75037596
Green H., and Kehinde, O. Cell 1, 113-116 1974
Green H., and Kehinde, O. Cell 5, 19-27 1975
Green H. Triglyceride-accumulating clonal cell line. U.S. Pat. 4,003,789 dated Jan. 18, 1977
|Date Created||07/17/2012 07:14 PM
|Date Updated||07/25/2012 05:22 PM