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NIH3T3 Cell Line

General Information
OrganismMus musculus, mouse
Cell Line DescriptionNIH 3T3 mouse embryonic fibroblast cells were initiated from a cell line isolated at the New York University School of Medicine Department of Pathology in 1962. 3T3 refers to the cell transfer and inoculation method for the line, and means “3-day transfer, inoculum 3 x 105 cells.” Using this method, 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 originally obtained the cells from desegregated NIH Swiss mouse embryo fibroblasts.
Product FormatFrozen
Growth ModeAdherent
Biosafety Level1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
ApplicationsDNA transfection studies
NIH 3T3 cell line has been used to:
  1. study calcium-mediated actin reset (CaAR) in response to physiological changes
  2. determine the viral titer of the retroviral construct -ST40hTNFα
  3. study the effects of activation of the transient receptor potential vanilloid 3 channel (TRPV3) on adipocyte differentiation
Shipped inDry ice
Storage Temperature−196°C
KaryotypeNot specified
ImagesNIH3T3 Cell Line
Products  Not specified
ReceptorsNot specified
MycoplasmaContamination was eliminated with Mycoplasma Removal Agent, then negative in DAPI, microbiological culture, PCR assays, RNA hybridization
CytogeneticsMurine hypertriploid karyotype with 3% polyploidy - 68(58-73)<3n>
VirusesELISA: reverse transcriptase negative; PCR: SMRV -
CommentsThe original cells are extremely contact inhibited, although the cell line is no longer inhibited. NIH 3T3 cells are receptive to transformation with SV40 and polyomavirus. Moreover, 3T3 cells are sensitive to sarcoma virus focus formation, leukemia virus and are inhibited by temazepam and other benzodiazepines.
Culture Conditions and Handling
Culture MediumDMEM + 2 mM Glutamine + 10% Calf Serum (CS).
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0-3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until the cell layer is dispersed.
  4. Add 6.0-8.0 mL of complete growth medium, and then aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to the new culture vessel.
  6. Incubate cultures at 37°C.
Split RatioSplit culture at 1:10 every 4 to 6 days using trypsin/EDTA
Medium RenewalTwice per week
Culture Temperature37°C
Incubation ConditionCarbon dioxide (CO2), 5%
Doubling TimeCa. 20 hours
HarvestCell harvest of about 20 x 106 cells/175 cm2 flask
StorageFrozen with 70% medium, 20% FBS, 10% DMSO

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For research use only. Not intended for any clinical use.

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