Crl 1772

Manufactured by American Type Culture Collection

Sourced in United States

The CRL-1772 is a laboratory equipment product from American Type Culture Collection. It is designed for cell culture applications. The core function of this product is to provide a controlled environment for the growth and maintenance of cells in vitro.

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ATCC® CRL-1772™). (23 citations) C2C12 myoblasts (CRL-1772) (4 citations) C2C12 (CRL-1772) (4 citations)

93 protocols using crl 1772

Cell Culture and Differentiation of C2C12 and PC12 Cells

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Cell culture of C2C12 mouse myoblasts (American Type Culture Collection (ATCC), CRL-1772) and PC12 rat adrenal gland pheochromocytoma cells (ATCC, CRL-1772) was performed as previously reported^{9 (link)–13 (link),15 (link)}. In brief, C2C12 cells were cultured in DMEM with 10% FBS and 1% penicillin–streptomycin (Invitrogen). Cells were seeded onto a 24-well plate at 50,000 cells per ml and incubated for 10–14 days in DMEM with 1% FBS and 1% penicillin–streptomycin to differentiate into myotubules. PC12 cells were grown in DMEM with 12.5% horse serum, 2.5% FBS and 1% penicillin–streptomycin. Cells were seeded onto a 24 well-plate, and 50 ng ml⁻¹ nerve growth factor (Invitrogen) was added 24 h after seeding.

Ji T., Li Y., Deng X., Rwei A.Y., Offen A., Hall S., Zhang W., Zhao C., Mehta M, & Kohane D.S. (2021). Delivery of local anaesthetics by a self-assembled supramolecular system mimicking their interactions with a sodium channel. Nature biomedical engineering, 5(9), 1099-1109.

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C2C12 Myoblast Encapsulation for Bioprinting

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C2C12 immortalized mouse myoblasts (CRL-1772TM, ATCC) were cultured in T75 and T175 flasks with proliferation medium: Dulbecco’s Modified Eagle Medium, high glucose, pyruvate (Gibco, 11995-081); qualified, heat-inactivated fetal bovine serum (Gibco, 16140-071) at 10% (v/v); and penicillin-streptomycin (Gibco, 15070-063) at 1% (v/v). When the cells reached above 80% confluency, they were suspended using a 0.05% of Trypsin-EDTA solution (Gibco, 15400054), diluted in DPBS, and reseeded into a new flask at an appropriate seeding density to enable cell expansion. Once an adequate number of cells were generated, the cells were again suspended, counted, and encapsulated in the gelatin bioink warmed to 37 °C at concentrations of 1, 10, and 30×10⁶ cells/mL for rheological and cell printing tests.

Schmid V.H., Cammarata K.V., Bruns B.U, & Schmidt G.W. (1997). In vitro reconstitution of the photosystem I light-harvesting complex LHCI-730: Heterodimerization is required for antenna pigment organization. Proceedings of the National Academy of Sciences of the United States of America, 94(14), 7667-7672.

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C2C12 Myoblast Differentiation Protocol

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C2C12 myoblast cells (ATCC^® CRL-1772 TM) were cultured in Dulbecco’s modified Eagle’s Medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% Fetal Bovine Serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA), 1% penicillin–streptomycin (Thermo Fisher Scientific, USA) in a humidified incubator at 37 °C and 5% CO₂. Subsequently, the cells were transferred to and cultured in 6-well plates (1 × 10⁵ cells/well). C2C12 myoblasts were collected or induced to undergo myogenic differentiation after transfections with mimic-miR-155-5p. For differentiation, once the myoblasts reached 80–90% confluence, the growth medium was replaced with FBS-free, DMEM-containing Horse Serum (2%), L-glutamine, and penicillin/streptomycin (1%) for 120 h. All experiments were performed in triplicate for each group.

Lopes L.O., Cury S.S., de Moraes D., Oliveira J.S., de Oliveira G., Cabral-Marques O., Fernandez G.J., Hirata M.H., Wang D.Z., Dal-Pai-Silva M., Carvalho R.F, & Freire P.P. (2024). The Impact of miR-155-5p on Myotube Differentiation: Elucidating Molecular Targets in Skeletal Muscle Disorders. International Journal of Molecular Sciences, 25(3), 1777.

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Culturing BJ and C2C12 Cells

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Human BJ fibroblasts (CRL-2522) and C2C12 mouse myoblasts (CRL-1772TM) were obtained from ATCC^Ⓡ. Culture media and reagents were purchased from (Invitrogen™, United States), unless indicated otherwise. BJ cells were cultured in Eagle’s minimum essential medium (EMEM). C2C12 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM). The culture media for both cell lines were supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% (v/v) penicillin-streptomycin. Cells were grown at 37°C in a 5% CO₂ atmosphere.

González-Gamboa I., Velázquez-Lam E., Lobo-Zegers M.J., Frías-Sánchez A.I., Tavares-Negrete J.A., Monroy-Borrego A., Menchaca-Arrendondo J.L., Williams L., Lunello P., Ponz F., Alvarez M.M, & Trujillo-de Santiago G. (2022). Gelatin-methacryloyl hydrogels containing turnip mosaic virus for fabrication of nanostructured materials for tissue engineering. Frontiers in Bioengineering and Biotechnology, 10, 907601.

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C2C12 Myotube Differentiation Assay

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Mouse C2C12 myoblasts (CRL-1772^TM, ATCC, Manassas, VA, USA) were maintained in DMEM supplemented with 10% FBS and 1% PS at 37 °C in a 5% CO₂ humidified incubator. When the confluence of the cells reached 80%, the culture medium was replaced with 2% horse serum (HS) containing DMEM to induce myotube differentiation, and the cells were incubated for 7 days with medium replacement every 24 h. The cells were then serum-starved overnight before PEG-BHD1028 or human gAcrp30 (gAD) treatment.

Lee I.K., Kim G., Kim D.H, & Kim B.B. (2021). PEG-BHD1028 Peptide Regulates Insulin Resistance and Fatty Acid β-Oxidation, and Mitochondrial Biogenesis by Binding to Two Heterogeneous Binding Sites of Adiponectin Receptors, AdipoR1 and AdipoR2. International Journal of Molecular Sciences, 22(2), 884.

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Culturing and Differentiating Cancer Cell Lines

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MDA-MB-231 breast cancer cells^{38 (link)} (HTB-26, ATCC, Manassas, VA), MCF-7 breast cancer cells^{39 (link)} (HTB-22, ATCC), ZR75-1 breast cancer cells^{40 (link)} (CRL-1500, ATCC), PC-3 prostate cancer cells^{41 (link)} (CRL-1435, ATCC), and RWGT2 lung cancer cells^{15 (link),42 (link)} (isolated from bone metastases by Dr. Guise as reported^{15 (link)}) were cultured in Dulbecco's modified Eagle's media (DMEM) (Hyclone, Logan, UT) containing 10% heat-inactivated fetal bovine serum (FBS) (Hyclone). JJN-3 multiple myeloma cells^{43 (link)} (ACC 541, DSMZ) were cultured in RPMI 1640 (Invitrogen, Grand Island, NY) containing 10% heat-inactivated FBS. A549 cancer cells (CCL-185, ATCC) were cultured in 1640 RPMI (Hyclone) containing 10% heat-inactivated FBS. C2C12 myoblast cells (CRL-1772, ATCC) were cultured subconfluently in DMEM containing 10% heat-inactivated FBS. C2C12 myoblasts were differentiated into myotubes by culture in DMEM containing 2% heat-inactivated horse serum (HS) (Hyclone). All cells were maintained at 37° C with 5% CO₂ in a humidified chamber. All cells were verified to be free of mycoplasma contamination via routine PCR testing. No independent verification was completed. Cells treated with recombinant human TGF-β1 (R&D Systems, Minneapolis, MN) were starved in DMEM (no serum) for 16-20 hrs and 5 ng/ml TGF-β1 was added to cells in DMEM.

Waning D.L., Mohammad K.S., Reiken S., Xie W., Andersson D.C., John S., Chiechi A., Wright L.E., Umanskaya A., Niewolna M., Trivedi T., Charkhzarrin S., Khatiwada P., Wronska A., Haynes A., Benassi M.S., Witzmann F.A., Zhen G., Wang X., Cao X., Roodman G.D., Marks A.R, & Guise T.A. (2015). Excess TGF-β mediates muscle weakness associated with bone metastases in mice. Nature medicine, 21(11), 1262-1271.

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Stem/progenitor cell culture protocol

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Two different types of cells were used to model stem/progenitor cells for in vitro studies. Multipotent mouse C2C12 myoblasts (ATCC® CRL-1772™, Manassas, VA, US) were cultured in Dulbecco's modified Eagle's media (DMEM, Thermo Fisher Scientific Inc.) with 10% fetal bovine serum (FBS; Thermo Fisher Scientific Inc.) and 1% penicillin-streptomycin (PS, 10,000 U/mL, Fisher Scientific). Poietics™ human mesenchymal stem cells (hMSCs) (PT-2501, Lonza Walkersville Inc. MD) were cultured with mesenchymal stem cell growth medium (MSCGM™ BulletKit™, PT-3001, Lonza Walkersville Inc. MD). All the cells were kept at 37 °C, 5% CO₂ in a humidified incubator. No mycoplasma contamination was found in cell cultures, which was monitored using a mycoplasma detection kit (Nanjing Vazyme Biotech Co., Ltd. China, MycoBlue, #D101-02) and Hoechst 33342 (Anaspec, USA) staining.

Zhang X., Li K., Wang C., Rao Y., Tuan R.S., Wang D.M, & Ker D.F. (2024). Facile and rapid fabrication of a novel 3D-printable, visible light-crosslinkable and bioactive polythiourethane for large-to-massive rotator cuff tendon repair. Bioactive Materials, 37, 439-458.

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C2C12 Mouse Myoblast Cell Culture

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C2C12 mouse myoblast cells were obtained from a commercial vendor (ATCC Cat# CRL-1772, RRID: CVCL_0188). Cells were cultured at 37°C with 5% CO₂ in a humidified incubator in media consisting of Dulbecco’s modified Eagle medium (DMEM; Gibco #11995-065) supplemented with 1% v/v penicillin-streptomycin (Gibco #15140-122) and either 10% v/v FBS (Gibco #A3161001) for proliferation media or 2% v/v horse serum (Gibco #16050-130) for differentiation media. Cells were grown in T75 flasks to ~80% confluence then washed with sterile PBS wash and detached (Gibco TrypLE Express #LTS12604021) for passaging. Cells used for all experiments were ≤9 passages from stock and were seeded into 6-well plates for experiments unless otherwise stated.

Trewin A.J., Silver J., Dillon H.T., Della Gatta P.A., Parker L., Hiam D.S., Lee Y.P., Richardson M., Wadley G.D, & Lamon S. (2022). Long non-coding RNA Tug1 modulates mitochondrial and myogenic responses to exercise in skeletal muscle. BMC Biology, 20, 164.

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C2C12 Cell Differentiation and Treatment

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C2C12 cells (passages 4–17, CRL-1772; ATCC, Manassas, VA, USA) were incubated at 37 °C in a 5% CO₂ atmosphere. The C2C12 cells were cultured in DMEM with 4500 mg/L D-glucose, 584 mg/L L-glutamine, 110 mg/L sodium pyruvate, 1200 mg/mL sodium bicarbonate, 100 Units/mL penicillin, 100 µg/mL streptomycin, and 10% FBS. For C2C12 differentiation, 9 × 10⁴ cells were seeded and then, cells at 70–80% confluence were replaced with the differentiation medium containing DMEM supplemented with 2% horse serum for 6 days. The C2C12 cells were treated with ECR (50 and 100 µg/mL) and DEX (39.2 µg/mL) for 24 h. The assessments of ECR on cell viability were performed using 3-(4,5-dimethylthiazol-2-yl) (MTT; Duchefa Biochemie, Haarlem, The Netherlands) as previously described [53 (link)].

Kim N.H., Lee J.Y, & Kim C.Y. (2023). Protective Role of Ethanol Extract of Cibotium barometz (Cibotium Rhizome) against Dexamethasone-Induced Muscle Atrophy in C2C12 Myotubes. International Journal of Molecular Sciences, 24(19), 14798.

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Culturing Primary Cells and Cell Lines

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Primary normal human dermal fibroblasts (HDMFs; ATCC® PCS-201-010™) were maintained in DMEM medium (GIBCO) containing 20% FBS (Seradigm), 0.2 mM L-Glutamine (Gibco), and 100 U/ml penicillin/streptomycin (Gibco). Murine myoblasts (C2C12; ATCC® CRL-1772) were cultured in DMEM medium + GlutaMAX™ (Gibco) containing 10% FBS (Seradigm) and 100 U/ml penicillin/streptomycin (Gibco). Primary patient-derived CMCs were propagated in Ham’s F12 medium (Gibco) supplemented with 10% FBS (Seradigm), 20 ng/ml Recombinant Human bFGF (PeproTech), 0.2 mM L-Glutamine (Gibco), 0.005 U/ml human Erythropoietin (Sigma), and 100 U/ml penicillin/streptomycin (Gibco). Human HEK293FT cells (Life Technologies) used in lentiviral production were grown in DMEM medium (Gibco) containing 10% FBS (Seradigm) and 0.2 mM L-Glutamine (Gibco). All cell lines were propagated under standard incubation conditions at 37°C with 5% atmospheric CO₂ and, further, were enzymatically passaged using TrypLE™ (Life Technologies) when approaching ≈70% confluence.

Moore JB I.V., Zhao J., Keith M.C., Amraotkar A.R., Wysoczynski M., Hong K.U, & Bolli R. (2016). The Epigenetic Regulator HDAC1 Modulates Transcription of a Core Cardiogenic Program in Human Cardiac Mesenchymal Stromal Cells (CMCs) through a p53-Dependent Mechanism. Stem cells (Dayton, Ohio), 34(12), 2916-2929.

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