General Information


NIH 3T3 mouse embryonic fibroblast cells come from a cell line isolated and initiated in 1962 at the New York University School of Medicine Department of Pathology. 3T3 refers to the cell transfer and inoculation protocol for the line, and means “3-day transfer, inoculum 3 x 10^5 cells.” Using this protocol, the immortal cell line begins to thrive and stabilize in cell culture after about 20-30 generations of in vitro growth. George Todaro and Howard Green, the scientists who first cultured this cell line, obtained the cells from desegregated NIH Swiss mouse embryo fibroblasts. The cell line has since become a standard fibroblast cell line.

Commercially available NIH3T3 transfection reagent is available from Altogen Biosystems.


Swiss 3T3 cells are inhibited by temazepam and other benzodiazepines. The original cells are extremely contact inhibited, although the cell line is no longer inhibited. 3T3 cells are sensitive to sarcoma virus focus formation, as well as leukemia virus.

3T3 cells are also receptive to transformation with polyomavirus and SV40.

NIH 3T3 cell cultureNIH 3T3 cell culture

Copyright picture from Reproduced with permission from Altogen Biosystems.


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3T3 mouse cells are hypertriploid. The modal chromosome number is 68, which occurs in 30% of cells. Higher ploidies occur at a much lower rate of 2.4%.


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Research Articles:

• Cytogenic characterization of the cell line: In this study, scientists studied the genome of NIH 3T3 cells. They characterized the gains and losses of copy numbers in NIH 3T3, and found that it had only gained four derivative chromosomes since its initial cytogenic study in 1989. Efforts to align NIH 3T3’s genome imbalances with homologous regions in humans indicated that imbalances in NIH 3T3 were similar to those in human solid cancers of the ectoderm.
• Comparison of NIH 3T3 cells to primary embryo cells: In an early study, the NIH 3T3 cells were compared to both other 3T3 cells and primary embryo cells. It was found that sensitivity to sarcoma virus formation and leukemia virus growth were comparable between NIH 3T3 cells and the other cells studied.
• Identification of Canine Distemper Virus (CDV) receptors for uptake: To determine what receptors CDV utilizes for uptake into susceptible cells, a monoclonal antibody was identified that binds to the cell surface and inhibits CDV infection. Once it was identified that CD9 was the receptor bound by this antibody, a plasmid was transfected into NIH 3T3 cells to express CD9. These cells were subsequently determined to be more permissive to CDV, however immunoprecipitation and virus-overlay protein binding assays were unable to determine if CDV is directly binding to the CD9 surface receptor.
• Mesothelioma glycoprotein antigen production: In this study, a cDNA fragment that encodes a glycoprotein common to mesothelial cells, mesothelioma and ovarian cancer cells was identified. The cDNA was then transfected into NIH 3T3 cells, which produced the glycoprotein on their surface. The glycoprotein was subsequently named mesothelin and may play a role in cellular adhesion.
• Reversible immortalization: NIH 3T3 cells were used as a control in a study of reversible immortalization of mammalian cells using an oncogene. Scientists were able to determine that the oncogene provided immortalization, and then, once excised, cells reverted to their pre-immortalization characteristics.