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3T3 cells

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NIH-3T3 fibroblasts in cell culture.

3T3 cells are several cell lines of mouse embryonic fibroblasts. The original 3T3 cell line (3T3-Swiss albino) was established in 1962 by two scientists then at the Department of Pathology in the New York University School of Medicine, George Todaro and Howard Green. Todaro and Green originally obtained their 3T3 cells from Swiss albino mouse embryo tissue.[1] Later, as a principal investigator position at the National Cancer Institute in Bethesda, Maryland, Todaro repeated the isolation procedure from the NIH Swiss mouse embryo with his students and established NIH-3T3 cell line.[2]


The '3T3' designation refers to the abbreviation of "3-day transfer, inoculum 3×105 cells." This cell line was originally established from the primary mouse embryonic fibroblast cells that were cultured by the designated protocol, so-called '3T3 protocol'. The primary mouse embryonic fibroblast cells were transferred (the "T") every 3 days (the first "3"), and inoculated at the rigid density of 3×105 cells per 20 cm2 dish (the second "3") continuously.[2] The spontaneously immortalized cells with stable growth rate were established after 20 to 30 generations in culture, and then named '3T3' cells. Since then, several cell lines have been established with this procotol:[3]

  • 3T3-Swiss albino, the original 1962 cell line
  • NIH-3T3, also from Swiss albino mice
  • BALB/c-3T3 clone A31, from BALB/c mice


Swiss 3T3 can be inhibited by temazepam and other benzodiazepines. These cells are also contact inhibited. The cells are sensitive to sarcoma virus and leukemia virus focus formation. 3T3 cells can be transformed with SV40 and some other polyomaviruses.[6]

Culture characteristics[edit]

Adherent cells grow as a monolayer. A confluent monolayer yields 40000 cells/cm2. [7]


Lysophosphatidylcholine (lyso-PC) induces AP-1 activity and c-jun N-terminal kinase activity (JNK1) by a protein kinase C-independent pathway. [8]


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%.[7]


  1. ^ Todaro, GJ; Green, H (1963). "Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines". J. Cell Biol. 17 (2): 299–313. doi:10.1083/jcb.17.2.299. PMC 2106200. PMID 13985244.
  2. ^ a b "Generating the 3T3 cell line, the oncogene hypothesis and horses". www.asbmb.org. Retrieved 2020-07-28.
  3. ^ Capes-Davis, Amanda; Freshney, R. Ian (2021). Freshney's Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications. John Wiley & Sons. p. 611. ISBN 978-1-119-51304-9.
  4. ^ Rheinwatd, James G.; Green, Howard (November 1975). "Seria cultivation of strains of human epidemal keratinocytes: the formation keratinizin colonies from single cell is". Cell. 6 (3): 331–343. doi:10.1016/S0092-8674(75)80001-8. PMID 1052771. S2CID 53294766.
  5. ^ Green, Howard; Kehinde, Olaniyi (March 1974). "Sublines of mouse 3T3 cells that accumulate lipid". Cell. 1 (3): 113–116. doi:10.1016/0092-8674(74)90126-3.
  6. ^ NIH 3T3 Cell Line Origins, Characteristics, Expression, and Cytogenetics
  7. ^ a b "3T3-Swiss albino ATCC CCL-92 Mus musculus embryo". www.atcc.org. Retrieved 2020-07-28.
  8. ^ Fang, X.; Gibson, S.; Flowers, M.; Furui, T.; Bast, R. C.; Mills, G. B. (1997-05-23). "Lysophosphatidylcholine stimulates activator protein 1 and the c-Jun N-terminal kinase activity". The Journal of Biological Chemistry. 272 (21): 13683–13689. doi:10.1074/jbc.272.21.13683. ISSN 0021-9258. PMID 9153219.

External links[edit]