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Nmumg cells

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NMuMG cells are a cell line derived from normal mouse mammary gland epithelial cells. They are a commonly used model system for studying mammary gland biology and function.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Recombinant activin a, by R&D Systems (2 mentions) Nih3t3 cells, by American Type Culture Collection (1 mentions) Penicillin streptomycin, by Caisson (1 mentions) Bovine calf serum (bcs), by Thermo Fisher Scientific (1 mentions) Dmem media, by Thermo Fisher Scientific (1 mentions) L glutamine, by Caisson (1 mentions) Penicillin streptomycin, by Sartorius (1 mentions) Recombinant tgf β, by Thermo Fisher Scientific (1 mentions) Nih3t3, by American Type Culture Collection (1 mentions) Nepa21 electroporator, by Nepa Gene (1 mentions) Tgf β1, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Insulin, by Roche (1 mentions) 293t cell, by American Type Culture Collection (1 mentions) Megm bullet kit, by Lonza (1 mentions)

8 protocols using nmumg cells

1

Culturing and Transfection of NMuMG Cells

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NMuMG cells (ATCC, Manassas, VA) were cultured using the manufacturer’s recommended culturing protocol with the addition of 1 mM Sodium Pyruvate and 100 U ml-1 penicillin, 100 μg ml-1 streptomycin (Thermo Fisher Scientific) to the media. These cells were authenticated by ATCC and found to be mycoplasma free by Hoechst DNA stain and agar culture. Transfection, selection and clonal isolation of NMuMG cells were performed similarly to CHO-K1 cells.
Nandagopal N., Santat L.A, & Elowitz M.B. (2019). Cis-activation in the Notch signaling pathway. eLife, 8, e37880.
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2

Genetic Models for Mammary Studies

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NMuMG cells were purchased from ATCC (CRL-1636). C57BL/6 itgβ4flox/flox mice were generated as described (13 (link)). C57BL/6 WAP-Cre mice (11 (link)) and B6.129 ROSA26mT/mG mice (12 ) were obtained from Ingolf Bach and Jaime Rivera, respectively at the University of Massachusetts Medical School (11 (link), 12 ). All of the animal breeding and procedures were approved by the Institutional Animal Care and Use Committee of the University of Massachusetts Medical School.
Walker M.R., Amante J.J., Li J., Liu H., Zhu L.J., Feltri M.L., Goel H.L, & Mercurio A.M. (2019). Alveolar Progenitor Cells in the Mammary Gland are Dependent on β4 Integrin. Developmental biology, 457(1), 13-19.
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3

Cell Culture Maintenance and Preparation

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NIH 3T3 cells (ATCC, USA) expressing H2B-mTurqoise2 and H2B-Citrine were kindly provided by Dr. Michael Elowitz (California Institute of Technology, USA). 3T3 cells were maintained in DMEM media (GIBCO, USA) supplemented with 10% BCS (GIBCO, USA), 1% l-glutamine (Caisson Laboratories, Inc., USA), and 1% penicillin/streptomycin (Caisson Laboratories, Inc., USA) at 37°C and 5% CO2. NMUMG cells (ATCC, USA) were maintained in DMEM media supplemented with 10% FBS (GIBCO, USA), 1% l-glutamine, and 1% penicillin/streptomycin at 37°C and 5% CO2. Prior to device seeding, cells were trypsinized and re-suspended in 500 μL of fresh media.
Curtis A., Li D.J., DeVeale B., Onishi K., Kim M.Y., Blelloch R., Laird D.J, & Hui E.E. (2017). Patterning of Sharp Cellular Interfaces With A Reconfigurable Elastic Substrate. Integrative biology : quantitative biosciences from nano to macro, 9(1), 50-57.
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4

Immortalized Wild-Type Mouse Cell Cultures

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WT MEFs (referred to in other publications as RPTPα+/+ cells) were isolated from embryonic day 13 (E13)–E15 WT embryos and underwent spontaneous immortalization (Su et al., 1999 (link)). NIH3T3 (#CRL-1658; ATCC), WT MDCK (#CCL-34; ATCC), and MDCK α-catenin KD (generated from the WT MDCK by shRNA [Benjamin et al., 2010 (link)]) were received from MBI Singapore cell collection. Vinculin−/− MEFs were obtained from Benny Geiger’s lab (Weizmann Institute of Science), originally generated from vinculin−/− mice provided by E.D. Adamson (Burnham Institute, La Jolla, CA; Xu et al., 1998 (link)). NMuMG cells (#CRL1636; ATCC) were received from Yaron Antebi’s lab (Weizmann Institute of Science). All cells were cultured at 37°C in a 5% CO2 incubator in DMEM supplemented with 10% FBS and 100 IU/ml penicillin-streptomycin (all reagents were from Biological Industries). Recombinant TGFβ (10 ng/ml) was purchased from Peprotech (#100-21). For EMT experiments, the cells were treated with TGFβ for 48–72 h. Transfections were carried out 1 d before measurements using the NEPA21 Electroporator (Nepa Gene) according to the manufacturer’s instructions, with ∼106 cells per reaction and 10 μg DNA.
Mukherjee A., Melamed S., Damouny-Khoury H., Amer M., Feld L., Nadjar-Boger E., Sheetz M.P, & Wolfenson H. (2022). α-Catenin links integrin adhesions to F-actin to regulate ECM mechanosensing and rigidity dependence. The Journal of Cell Biology, 221(8), e202102121.
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5

TGFβ1 Signaling in NMuMG Cells

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NMuMG cells (ATCC, Manassas, Virginia, United States) were maintained at 37°C and 5% CO2 in DMEM (Dulbecco's Modification of Eagle's Medium), supplemented with 10% fetal bovine serum (FBS) (Invitrogen), 10μg/mL insulin (Roche), 2mM L-glutamine (Invitrogen), and 200 U mL-1 penicillin/streptomycin (Invitrogen). All experiments were performed in medium as described above but without the added insulin. The cells were serum-starved overnight prior to treatment with TGFβ1. For transfection, NMuMGs were seeded at 3 × 106 cells on a 10cm plate and grown overnight to 70-90% confluency. The cells were transfected with Lipofectamine 3000™ Reagent in serum-free media, adjusted for a 10 cm plate according to the manufacturer's instructions. The transfection mix was then added to the adherent cells and topped up with 10% FBS DMEM after 3 hours. The cells were harvested 48 hours following transfection. TGFβ1 (Sigma-Aldrich) was added to the cells at a concentration of 2ng/mL in serum-free media (1X DMEM, Corning, USA) in all experiments.
Conway J., Al-Zahrani K.N., Pryce B.R., Abou-Hamad J, & Sabourin L.A. (2017). Transforming growth factor β-induced epithelial to mesenchymal transition requires the Ste20-like kinase SLK independently of its catalytic activity. Oncotarget, 8(58), 98745-98756.
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6

Comprehensive Breast Cancer Cell Culture

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All human breast cancer cells, NMuMG cells, and 293T cells were obtained from American Type Culture Collection. HMLE cells were obtained from R. Weinberg (MIT, USA). MDAMB231, NMuMG, and 293T cells were maintained in DMEM supplemented with 10% FBS. MCF7 cells were maintained in DMEM supplemented with 10% FBS and 10 μg/ml insulin. BT474, HCC1143, BT549, and MDAMB468 cells were maintained in RPMI1640 supplemented with 10% FBS. SUM149 and SUM159 cells were maintained in Ham’s F12 supplemented with 5% FBS, 10 mM HEPES, 1 µg/ml Hydrocortisone, and 5 μg/ml Insulin. MCF10A and MCF12A cells were maintained in MEGM supplemented 100 ng/ml cholera toxin. HMLE cells were maintained in MEGM bullet kit (Lonza, no. CC-3150). All media were supplemented with 1% penicillin and streptomycin.
Okada N., Ueki C., Shimazaki M., Tsujimoto G., Kohno S., Muranaka H., Yoshikawa K, & Takahashi C. (2023). NFYA promotes malignant behavior of triple-negative breast cancer in mice through the regulation of lipid metabolism. Communications Biology, 6, 596.
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7

Epithelial-Mesenchymal Transition Protocols

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NMuMG cells, obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and the 4T1 progression model, acquired from Dr. William Schiemann, were cultured in DMEM high glucose supplemented with 10% FBS and 1% antibiotic/antimycotic solution (penicillin G, streptomycin, amphotericin B). HMLE cells were obtained from Dr. Sendurai Mani and cultured in DMEM:F12 supplemented with 5% calf serum, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 20 ng/ml EGF and 1% antibiotic/antimycotic. The HMLE subpopulations were initially isolated as described previously (4 (link), 32 (link)) and subsequently FACS sorted using CD44 and CD24 antibodies (see supplementary methods). All cells were cultured in a 37°C, 5% CO2 incubator. Recombinant activin A was purchased from R&D systems (Minneapolis, MN, USA). TGFβ2 was a generous gift from Genzyme Corporation (Cambridge, MA, USA).
Howley B.V., Hussey G.S., Link L.A, & Howe P.H. (2015). Translational regulation of Inhibin βA by TGFβ via the RNA-binding protein hnRNP E1 enhances the invasiveness of epithelial-to-mesenchymal transitioned cells. Oncogene, 35(13), 1725-1735.
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8

Epithelial-Mesenchymal Transition Protocols

Cited 1 time
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NMuMG cells, obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and the 4T1 progression model, acquired from Dr. William Schiemann, were cultured in DMEM high glucose supplemented with 10% FBS and 1% antibiotic/antimycotic solution (penicillin G, streptomycin, amphotericin B). HMLE cells were obtained from Dr. Sendurai Mani and cultured in DMEM:F12 supplemented with 5% calf serum, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 20 ng/ml EGF and 1% antibiotic/antimycotic. The HMLE subpopulations were initially isolated as described previously (4 (link), 32 (link)) and subsequently FACS sorted using CD44 and CD24 antibodies (see supplementary methods). All cells were cultured in a 37°C, 5% CO2 incubator. Recombinant activin A was purchased from R&D systems (Minneapolis, MN, USA). TGFβ2 was a generous gift from Genzyme Corporation (Cambridge, MA, USA).
Howley B.V., Hussey G.S., Link L.A, & Howe P.H. (2015). Translational regulation of Inhibin βA by TGFβ via the RNA-binding protein hnRNP E1 enhances the invasiveness of epithelial-to-mesenchymal transitioned cells. Oncogene, 35(13), 1725-1735.
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