4T1 cell culture - 50% confluence

4T1 is a breast cancer cell line derived from the mammary gland tissue of a mouse BALB/c strain.[1] 4T1 cells are epithelial and are resistant to 6-thioguanine.[1] In preclinical research, 4T1 cells have been used to study breast cancer metastasis as they can metastasize to the lung, liver, lymph nodes, brain and bone.[2][3] The cells are known to be highly aggressive in live tissues.[4]

History

4T1 cell line was originally isolated by Fred Miller and colleagues as one of four sublines derived from the 410.4 tumor that was isolated from a single spontaneously arising mammary tumor.[5]

Characteristics

4T1 resembles human metastatic triple-negative breast cancer (TNBC) in lack of the expression of estrogen receptor (ER), progesterone receptor (PR), and epidermal growth factor receptor 2 (HER2).[6] In addition, it was described, that this cell line is poorly immunogenic in mice, which corresponds to the characteristics of human mammary adenocarcinoma and leads to the higher tumorigenicity and invasiveness of this breast cancer model.[1]

Application

To understand the pathogenesis of cancer patients, it is necessary to have model systems that faithfully mimic this condition. The application of a murine cancer cell line, such as 4T1, in a mouse model is of great value for preclinical TNBC studies. The 4T1 cell line is widely used as a syngeneic model for human triple-negative breast cancer, which is responsible for more than 17% of breast cancers diagnosed worldwide each year.[7] 4T1 cells can be transplanted into the fat pad of the murine mammary gland, where they are highly tumorigenic, invasive, and spontaneously metastasize to distant organs such as the lungs, liver, lymph nodes, brain, and bone.[1]

Cultivation

4T1 cells grow in 37 °C with 95% air and 5% of carbon dioxide (CO
2). Their average doubling time is 14 hours. The base medium for this cell line is RPMI-1640 Medium with a fetal bovine serum in a final concentration of 10%. 4T1 cells should not be allowed to become 100% confluent. Subculturing at 80% confluence is recommended. For detaching the cells from the surface, rinsing with 0.25% trypsin-0.53mM EDTA solution at room temperature is used. Complete growth medium 95% with DMSO 5% is used as a freezing medium and cells are stored at liquid nitrogen vapor phase temperature.[8]

Variants

Available variants
CVCL_0125 (4T1)
CVCL_QZ56 (4T1-eGFP-Puro)
CVCL_QZ59 (4T1-Fluc-Neo/iRFP-Puro)
CVCL_L899 (4T1-luc2)
CVCL_QZ61 (4T1-mNIS (monoclonal))
CVCL_QZ64 (4T1-mNIS-Puro)
CVCL_5I85 (4T1-Red-FLuc-GFP)
CVCL_GR31 (4T1.13)
CVCL_XG69 (4T1/GFP/FlucII)
CVCL_QZ57 (4T1-Fluc-Neo)
CVCL_QZ60 (4T1-iRFP-Puro)
CVCL_5J28 (4T1-luc2-GFP)
CVCL_QZ62 (4T1-mNIS-Neo/eGFP-Puro)
CVCL_QZ65 (4T1-mNIS-Puro/Fluc-Neo)
CVCL_HE47 (4T1-S)[9]
CVCL_GR32 (4T1.2)
CVCL_QZ58 (4T1-Fluc-Neo/eGFP-Puro)
CVCL_J239 (4T1-Luc)
CVCL_5J46 (4T1-luc2-tdTomato)
CVCL_QZ63 (4T1-mNIS-Neo/iRFP-Puro)
CVCL_5I84 (4T1-Red-FLuc)
CVCL_5J45 (4T1-tdTomato)
CVCL_JG34 (4T1/CMV-Luc #6)

[10]

References

  1. 1 2 3 4 Pulaski, BA; Ostrand-Rosenberg, S (May 2001). "Mouse 4T1 breast tumor model". Current Protocols in Immunology. 20 (1): Unit 20.2. doi:10.1002/0471142735.im2002s39. PMID 18432775. S2CID 205154914.
  2. ↑ Lelekakis, Maria; Moseley, Jane M.; Martin, T. John; Hards, Daphne; Williams, Elizabeth; Ho, Patricia; Lowen, Darren; Javni, Jeannie; Miller, Fred R.; Slavin, John; Anderson, Robin L. (1999). "A novel orthotopic model of breast cancer metastasis to bone". Clinical & Experimental Metastasis. 17 (2): 163–170. doi:10.1023/A:1006689719505. PMID 10411109. S2CID 2971601.
  3. ↑ Shengyu, Yang; et al. (2012). "Mouse Models for Tumor Metastasis". Rational Drug Design. Methods in Molecular Biology. Vol. 928. pp. 221–228. doi:10.1007/978-1-62703-008-3_17. ISBN 978-1-62703-007-6. PMC 3674868. PMID 22956145.
  4. ↑ Disch, Bryan; et al. (30 May 2014). "Development and Characterization of a Preclinical Model of Breast Cancer Lung Micrometastatic to Macrometastatic Progression". PLoS ONE. 9 (5): e98624. Bibcode:2014PLoSO...998624B. doi:10.1371/journal.pone.0098624. PMC 4039511. PMID 24878664.
  5. ↑ Aslakson, Cheryl J.; Rak, Janusz W.; Miller, Bonnie E.; Miller, Fred R. (1991-02-01). "Differential influence of organ site on three subpopulations of a single mouse mammary tumor at two distinct steps in metastasis". International Journal of Cancer. 47 (3): 466–472. doi:10.1002/ijc.2910470327. ISSN 0020-7136. PMID 1993557. S2CID 7663566.
  6. ↑ Schrörs, Barbara; Boegel, Sebastian; Albrecht, Christian; Bukur, Thomas; Bukur, Valesca; Holtsträter, Christoph; Ritzel, Christoph; Manninen, Katja; Tadmor, Arbel D.; Vormehr, Mathias; Sahin, Ugur (2020). "Multi-Omics Characterization of the 4T1 Murine Mammary Gland Tumor Model". Frontiers in Oncology. 10: 1195. doi:10.3389/fonc.2020.01195. ISSN 2234-943X. PMC 7390911. PMID 32793490.
  7. ↑ Anders, Carey K.; Carey, Lisa A. (June 2009). "Biology, Metastatic Patterns, and Treatment of Patients with Triple-Negative Breast Cancer". Clinical Breast Cancer. 9 (Suppl 2): S73–S81. doi:10.3816/CBC.2009.s.008. PMC 2919761. PMID 19596646.
  8. ↑ "4T1 ATCC CRL-2539 Mus musculus mammary gland This tumor p". www.lgcstandards-atcc.org. Retrieved 2020-09-08.
  9. ↑ Abe, Hirotake; Wada, Haruka; Baghdadi, Muhammad; Nakanishi, Sayaka; Usui, Yuu; Tsuchikawa, Takahiro; Shichinohe, Toshiaki; Hirano, Satoshi; Seino, Ken-ichiro (April 2016). "Identification of a highly immunogenic mouse breast cancer sub cell line, 4T1-S". Human Cell. 29 (2): 58–66. doi:10.1007/s13577-015-0127-1. hdl:2115/64955. ISSN 1749-0774. PMID 26857856. S2CID 16932816.
  10. ↑ "Cellosaurus cell line 4T1 (CVCL_0125)". web.expasy.org. Retrieved 2020-09-10.
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