L6 rat myoblasts

Manufactured by American Type Culture Collection

Sourced in United States

L6 rat myoblasts are a cell line derived from the skeletal muscle of a rat. They are commonly used as a model system for studying muscle cell biology and differentiation.

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Lab products found in correlation

Dmem high glucose, by Merck Group (2 mentions) Mouse c2c12 myoblast, by American Type Culture Collection (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Penicillin, by American Type Culture Collection (1 mentions) Streptomycin, by American Type Culture Collection (1 mentions) Mc3t3 e1 pre osteoblasts, by American Type Culture Collection (1 mentions) α mem, by Thermo Fisher Scientific (1 mentions) Penicillin, by Gemini Bio (1 mentions) Streptomycin, by Gemini Bio (1 mentions) Bovine calf serum (bcs), by Thermo Fisher Scientific (1 mentions) 3t3 l1 mouse embryonic fibroblasts, by American Type Culture Collection (1 mentions) Crl 1458, by American Type Culture Collection (1 mentions) Hepg2 human hepatocarcinoma cells, by American Type Culture Collection (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Tumor necrosis factor α tnf α, by Thermo Fisher Scientific (1 mentions) Penicillin a, by Thermo Fisher Scientific (1 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

L6 rat skeletal muscle myoblasts L6 myoblasts

13 protocols using l6 rat myoblasts

Expansion and Culture of Diverse Cell Lines

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Human bone marrow-derived MSCs from a single donor were purchased from RoosterBio (Frederick, MD). Cells were expanded in α-MEM (Invitrogen, Carlsbad, CA) supplemented with 10% v/v fetal bovine serum (FBS, JR Scientific, Woodland, CA), 100 units/mL penicillin, and 100 μg/mL streptomycin (Gemini Bio-Products, Sacramento, CA) under standard culture conditions until use at passage 4-5. MC3T3-E1 pre-osteoblasts (ATCC, Manassas, VA) and rat bone marrow stromal cells (rBMSCs) (Cyagen Biosciences Inc, Santa Clara, CA) were cultured in similar conditions. L6 rat myoblasts (ATCC) were maintained in DMEM supplemented with 10% v/v FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin under standard culture conditions. Myoblasts were kept below 70% confluency to prevent differentiation and myofibril formation.

Saiz AM J.r., Gionet-Gonzales M., Lee M.A, & Leach J.K. (2019). Conditioning of Myoblast Secretome using Mesenchymal Stem/Stromal Cell Spheroids Improves Bone Repair. Bone, 125, 151-159.

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Culturing and Differentiating Mouse and Rat Myoblasts

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C2C12 mouse myoblasts, L6 rat myoblasts, and 3T3-L1 mouse embryonic fibroblasts were purchased from ATCC (Cat#. CRL-1772, CRL-1458, Cat#. CL-173; Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) + 20% FBS, MEM +10% FBS, and DMEM + 10% Bovine Calf Serum (pre-adipocyte expansion medium), respectively (FBS; ThermoFisher Scientific, Waltham, MA, USA) at 37° C in 5% CO₂. To differentiate C2C12 and L6 myoblasts to myotubes, media was replaced with DMEM + 2% horse serum every 24 h for 6-7 days until cells were fully differentiated. To differentiate 3T3-L1 pre-adipocytes, pre-adipocyte expansion medium was replaced with differentiation medium (DMEM,10% FBS, 1μM Dexamethasone, 0.5mM IBMX, 1μg/mL insulin) for 48 h, then replaced with adipocyte maintenance medium (DMEM 10% FBS, 1μg/mL insulin) every 2 days for 7 days. HEK293 cells were cultured in high glucose DMEM (Sigma) supplemented with 10% fetal bovine serum at 37° C in 5% CO2. For transient transfection of the wild type (WT) and K14Q MOTS-c in pcDNA3.1(+), lipofectamine 3000 was used (ThermoFisher Scientific).

Zempo H., Kim S.J., Fuku N., Nishida Y., Higaki Y., Wan J., Yen K., Miller B., Vicinanza R., Miyamoto-Mikami E., Kumagai H., Naito H., Xiao J., Mehta H.H., Lee C., Hara M., Patel Y.M., Setiawan V.W., Moore T.M., Hevener A.L., Sutoh Y., Shimizu A., Kojima K., Kinoshita K., Arai Y., Hirose N., Maeda S., Tanaka K, & Cohen P. (2021). A pro-diabetogenic mtDNA polymorphism in the mitochondrial-derived peptide, MOTS-c. Aging (Albany NY), 13(2), 1692-1717.

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Cell Culture Protocol for Various Cell Lines

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C2C12 mouse myoblasts, L6 rat myoblasts, and HepG2 human hepatocarcinoma cells were obtained from the American Type Culture Collection (ATCC). Human fibroblasts were isolated from a skin biopsy of a healthy person (wild-type, WT) or MELAS patients harboring a heteroplasmic A3243G mutation. Cells were cultured in Dulbecco's modified Eagle's medium (25 mM glucose) supplemented with 10% fetal bovine serum in an atmosphere containing 5% CO₂ at 37°C.

Seo K.S., Kim J.H., Min K.N., Moon J.A., Roh T.C., Lee M.J., Lee K.W., Min J.E, & Lee Y.M. (2018). KL1333, a Novel NAD+ Modulator, Improves Energy Metabolism and Mitochondrial Dysfunction in MELAS Fibroblasts. Frontiers in Neurology, 9, 552.

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Rat Myoblast Differentiation Protocol

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L6 rat myoblasts were obtained from the American Type Culture Collection. Cells (5.0 × 10⁵) were seeded in 10‐cm plates in growth medium (GM) (AMEM supplemented with 10% fetal bovine serum and 1% antibiotic‐antimycotic agents) and incubated at 37°C and 5% CO₂. To initiate differentiation, cells were first grown to ~90% confluency in 6‐well plates and then shifted into differentiation medium (DM) consisting of AMEM, 1% antibiotic‐antimycotic agents, and 2% horse serum. DM was replaced every 24 hr.

Dhanani Z.N., Mann G, & Adegoke O.A. (2019). Depletion of branched‐chain aminotransferase 2 (BCAT2) enzyme impairs myoblast survival and myotube formation. Physiological Reports, 7(23), e14299.

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Differentiation and Treatment of Rat Myoblasts

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L6 rat myoblasts were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were grown in Dulbecco’s modified Eagle’s medium (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (Gibco, Gaithersburg, MD, USA) and 1% antibiotics (100 U/mL penicillin A and 100 mg/mL streptomycin; Gibco) at 37°C in an atmosphere of 5% CO₂. After cells reached approximately 80% confluence in plates, the growth medium was changed to a differentiation medium containing 2% horse serum (HyClone) and 1% antibiotics. During the 6 days of differentiation, the medium was changed every 2 days. When cells were fully differentiated, cells were treated with 50 ng/mL tumor necrosis factor-α (TNF-α; PeproTech, Rocky Hill, NJ, USA), CME (1 and 10 μg/mL; denoted as CME 1 and 10, respectively) and IcA (1 and 10 μM; denoted IcA 1 and 10, respectively) overnight.

Kwon D., Kim C., Woo Y.K, & Hwang J.K. (2021). Inhibitory Effects of Chrysanthemum (Chrysanthemum morifolium Ramat.) Extract and Its Active Compound Isochlorogenic Acid A on Sarcopenia. Preventive Nutrition and Food Science, 26(4), 408-416.

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Myoblast Differentiation and TNF-α Treatment

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L6 rat myoblasts were purchased from the American Type Culture Collection (USA) and cultured at 37°C and 5% CO₂ in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, USA) containing 10% fetal bovine serum (FBS; Hyclone). Myoblast differentiation into myotubes was performed according to a previous study [6 (link)]. Briefly, to induce differentiation, 10% FBS (Hyclone) in DMEM (Hyclone) was exchanged with 2% horse serum (Hyclone) in DMEM. This medium was replaced every two days for 6 days. On day 6, the cells were cotreated with both 50 ng/ml TNF-α (PeproTech, USA) and LJE or leonurine for 12 h.

Lee J., Kim C., Lee, H, & Hwang J.K. (2020). Inhibitory Effects of Standardized Leonurus japonicus Extract and Its Bioactive Leonurine on TNF-α-Induced Muscle Atrophy in L6 Myotubes. Journal of Microbiology and Biotechnology, 30(12), 1896-1904.

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Neuromuscular Junction Assay Protocol

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Neuromuscular junction (NMJ) assays were performed similarly to as described in previous studies (Harper et al., 2004 (link); Miles et al., 2004 (link)). Briefly, rat L6 myoblasts (ATCC CRL-1458) were seeded into Matrigel-coated Lab-Tek II 4-chambered cover glass and grown to confluency in DMEM/F-12, 10% FBS, 1× P-S medium. At 100% confluency, myoblasts were cultured in N2B27 medium supplemented with 100 nM RA, 200 nM Hh-Ag1.5 to drive multinucleated skeletal myotube formation. Twenty-four hours NS (day 38) were seeded onto myotubes at 5–8 NS per well in TDM and cultured for four days prior to fixation and fluorescence microscopy (day 42 SMNs).

Olmsted Z.T., Stigliano C., Marzullo B., Cibelli J., Horner P.J, & Paluh J.L. (2022). Fully Characterized Mature Human iPS- and NMP-Derived Motor Neurons Thrive Without Neuroprotection in the Spinal Contusion Cavity. Frontiers in Cellular Neuroscience, 15, 725195.

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Rat L6 myoblasts differentiation

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Rat L6 myoblasts (ATCC, Manassas, VA, USA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM 4.5 g/l glucose, Lonza, Basel, Switzerland) supplemented with 10% FBS (Hyclone, Pasching, Austria), 2 mM L-glutamine (Lonza) and 1% penicillin/streptomycin (Lonza) at +37°C in a humidified atmosphere of 5% CO
₂. The cells were seeded in multiwell plates at 2 × 10
⁴ cells/cm
² one day before starting the differentiation. Myoblasts were differentiated into myotubes by switching into α-MEM media (Gibco, Paisley, UK) supplemented with 2% horse serum (Gibco) and 1% penicillin/streptomycin. GR24 (3E,3aR,8bS)-3-[[(2 S)-4-methyl-5-oxo-2H-furan-2-yl] oxymethylidene]-4,8b-dihydro-3aH-indeno[1,2-b]furan-2-one, Chiralix, Nijmegen, Netherlands) was dissolved in DMSO. The control samples had equivalent DMSO concentration.

Modi S.R, & Kokkola T. (2018). Strigolactone GR24 upregulates target genes of the cytoprotective transcription factor Nrf2 in skeletal muscle. F1000Research, 7, 1459.

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Protein Extraction and Characterization in Rat L6 Myoblasts

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α-Glucosidase, p-nitrophenol- α-D glucopyranoside, and the primers of PTP1B, GLUT4, and β-actin were purchased from Sigma-Aldrich (St. Louis, USA). PTP1B (human recombinant) was purchased from Biomol International LP (PA, USA). TRIzol Reagent and M-PER^™ Mammalian Protein Extraction Reagent were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Primary antibodies, rabbitanti-GLUT4 and goat-anti-PTP1B, were purchased from Sino Biological (China). All other chemicals and reagents used were of analytical grade and purchased from VWR (Radnor, PA, USA).
Rat L6 myoblasts purchased from ATCC (Manassas, VA) were cultured in complete growth media (DMEM supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin, and 10 % FBS) at 37 °C in 5 % CO₂. L6 myoblasts were differentiated by the method described previously [21 (link), 22 (link)] with slight modification. Briefly, the medium was replaced with the DMEM supplemented with 2% FBS to induce differentiation when the cells reached 80–90% confluence. After 48 hours, the differentiated myotubes were serum-starved in 0.2 % BSA for 18 hours prior to the cell-based assays, except cytotoxicity assay.

Zehra S.A., Bhattarai P., Zhang J., Liu Y., Parveen Z., Sajid M, & Zhu L. (2022). In Vitro and In Vivo Evaluation of the Antidiabetic Activity of Solidago virgaurea Extracts. Current bioactive compounds, 19(4), e150622206034.

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Differentiation of Rat L6 Myoblasts

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Rat L6 myoblasts were obtained from American Type Culture Collection (Rockville, MD, USA) and maintained at 37 °C under a humidified 5% CO₂ atmosphere. Cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco BRL, NY, USA) supplemented with 10% fetal bovine serum (Gibco BRL) and 1% antibiotics (Gibco BRL). L6 myoblasts differentiation was induced by switching confluent cells to medium supplemented with 2% FBS. Experiments were performed in differentiated myoblasts.

Leem K.H., Kim M.G., Hahm Y.T, & Kim H.K. (2016). Hypoglycemic Effect of Opuntia ficus-indica var. saboten Is Due to Enhanced Peripheral Glucose Uptake through Activation of AMPK/p38 MAPK Pathway. Nutrients, 8(12), 800.

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