Nih 3t3

Manufactured by Lonza

Sourced in United States

The NIH-3T3 is a cell line derived from mouse fibroblast cells. This cell line is commonly used in various cell-based assays and research applications.

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5 protocols using nih 3t3

Cell Line Characterization and Use

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The cell line HEK-293T (Homo sapiens), ATCC, cat no. CRL-3216, cells were used by passage 7 in all experiments for this paper, mycoplasma-free (Lonza, LT07-118, control: LT07-518). The cell line NIH-3T3 (Mus muscles), ATCC, cat no. CRL-1658, used by passage 5 in all experiments for this paper, mycoplasma-free (Lonza, LT07-118, control: LT07-518)

Crapster J.A., Rack P.G., Hellmann Z.J., Le A.D., Adams C.M., Leib R.D., Elias J.E., Perrino J., Behr B., Li Y., Lin J., Zeng H, & Chen J.K. (2020). HIPK4 is essential for murine spermiogenesis. eLife, 9, e50209.

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In Vitro Fibroblast Proliferation Assay

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Human dermal fibroblasts (MRC-5) and murine embryo fibroblasts (NIH-3T3) were purchased from Lonza and grown in EMEM and DMEM, respectively, containing 10% FBS, pen/strep and L-Glut (Lonza, Walkersville, MD). Cell cultures were maintained at 37 °C in a humidified atmosphere with 5% CO₂. Cell cultures were analyzed for cell shape and growth changes and phase-contrast images were recorded with EVOS digital inverted microscope (Advanced Microscopy Group, Bothell, WA).
HelixComplex preparations were used for in vitro treatments of cell cultures using a range of concentrations previously determined in dose-response assays (4–400 μg/ml). Glycolic acid was used as fibroblast proliferation-inducer positive control at the concentration of 0.1 mM^{11 (link)}. For growth and proliferation assays, fibroblasts were seeded and treated when reached 50–60% of confluence (using a plating density of 10⁴ cells/ml).

Trapella C., Rizzo R., Gallo S., Alogna A., Bortolotti D., Casciano F., Zauli G., Secchiero P, & Voltan R. (2018). HelixComplex snail mucus exhibits pro-survival, proliferative and pro-migration effects on mammalian fibroblasts. Scientific Reports, 8, 17665.

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Cell Line Cultivation and Characterization

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NCI-H1993 (CRL-5909), NCI-H441 (HTB-174), SKBR3 (HTB-30), MKN-45 (ACC409), OE33 (ACC706), MCF7 (HTB-22), MDA-MB-231 (HTB-26) and NIH3T3 (CRL-1658) cell lines were obtained from ATCC (http://www.lgcstandards-atcc.org/en.aspx) or DSMZ (http://www.dsmz.de/), tested for mycoplasma infection on a regular basis using a commercial biochemical test (Lonza) and authenticated using STR profiling. All cells were cultured in Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham, 1:1 mixture supplemented with 10% fetal calf serum (FCS), or in case of MKN45 with 20% FCS. Upon serum withdrawal, FCS was replaced by 0.1% bovine serum albumin.

Schmidt U., Heller G., Timelthaler G., Heffeter P., Somodi Z., Schweifer N., Sibilia M., Berger W, & Csiszar A. (2021). The FAM3C locus that encodes interleukin-like EMT inducer (ILEI) is frequently co-amplified in MET-amplified cancers and contributes to invasiveness. Journal of Experimental & Clinical Cancer Research : CR, 40, 69.

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Cell Culture Conditions for KATO III and NIH/3T3

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Human gastric carcinoma cell line KATO III mouse embryo fibroblast cell line NIH/3T3 obtained from the American Type Culture Collection (ATCC) was maintained in Iscove's Modified Dulbecco's Medium, supplemented with 20% heat inactivated fetal bovine serum (Lonza, Switzerland), 100 IU/ml penicillin and streptomycin (Invitrogen, USA) and mouse embryo fibroblast cell line NIH/3T3 was maintained in Dulbecco's Modified Eagle Medium (Lonza, Switzerland), supplemented with 10% heat inactivated fetal bovine serum (Invitrogen, USA), 100 IU/ml penicillin and streptomycin (Lonza, Switzerland). All cultures were kept at 37°C in 5% CO₂ atmosphere.

Alshaer W., Ababneh N., Hatmal M., Izmirli H., Choukeife M., Shraim A., Sharar N., Abu-Shiekah A., Odeh F., Al Bawab A., Awidi A, & Ismail S. (2017). Selection and targeting of EpCAM protein by ssDNA aptamer. PLoS ONE, 12(12), e0189558.

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Cell Culture and Transduction Protocols

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HEK-293 (CRL-1573), HeLa cells (CCL-2), A431 (CRL-1555) and NIH/3T3 (CRL-1568) were cultured in Dulbecco's modified Eagle's medium (DMEM) (Lonza, Walkersville, MD, USA) supplemented with 2 mM L-glutamine, 10% (vol/vol) heat inactivated Fetal Calf Serum (FCS), and antibiotics (100 units/mL penicillin, 100 μg/mL streptomycin) (all from Life Technologies, Carlsbad, CA, USA) referred as to DMEM complete medium (DCM), unless otherwise stated. Jurkat clone E6–1 cells (TIB-152) were maintained in RPMI-1640 (Lonza) supplemented with 2 mM L-glutamine, heat-inactivated 10% FCS, and antibiotics, referred as to RPMI complete medium (RCM). All of these cell lines were obtained from the American Type Culture Collection (Rockville, MD, USA). The human Jurkat J77 cell line³⁷ (link) was maintained in RCM. 3T3, HeLa and A431 cells were infected with lentivirus encoding the firefly luciferase (Luc) gene^38,39 (link) has been described previously.⁴⁰ (link) The 3T3 cells stably expressing human EGFR (3T3-EGFR) were kindly provided by Antonio Villalobo (Instituto de Investigaciones Biomédicas Albert Sols, CSIC-UAM, Madrid, Spain) and cultured in DCM. All cell lines were routinely screened for the absence of mycoplasma contamination by PCR using the Mycoplasma Plus TM Primer Set (Stratagene, Cedar Creek, TX, USA).

Harwood S.L., Alvarez-Cienfuegos A., Nuñez-Prado N., Compte M., Hernández-Pérez S., Merino N., Bonet J., Navarro R., Van Bergen en Henegouwen P.M., Lykkemark S., Mikkelsen K., Mølgaard K., Jabs F., Sanz L., Blanco F.J., Roda-Navarro P, & Alvarez-Vallina L. (2017). ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy. Oncoimmunology, 7(1), e1377874.

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