C2C12 myoblasts (American Type Culture Collection, USA) were cultured in DMEM supplemented with 10% FBS and 1% P/S. The C2C12 myoblasts were trypsinized using 0.25% trypsin/0.1% EDTA when they reached 70–80% confluence. The cells were maintained in a cell culture incubator (Sanyo, Japan) with 5% CO2 at 37°C.
C2c12 myoblast
Manufactured by American Type Culture Collection
Sourced in United States, Germany, China, France
C2C12 myoblasts are an immortalized mouse skeletal muscle cell line. They can be used for studies of myogenesis and muscle cell differentiation.
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Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (39 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (38 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (27 mentions)
Horse serum, by Thermo Fisher Scientific (17 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (4 mentions)
Hek293t cell, by American Type Culture Collection (4 mentions)
Glutamax, by Thermo Fisher Scientific (4 mentions)
Hepes, by Thermo Fisher Scientific (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
Fetalplex, by Gemini Bio (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Hek293t, by American Type Culture Collection (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Merck Group (3 mentions)
Streptomycin, by Merck Group (3 mentions)
Glutamax 1, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Nih 3t3 fibroblasts, by American Type Culture Collection (2 mentions)
138 protocols using c2c12 myoblast
1
Culturing C2C12 Myoblasts for Research
2
Lentiviral Knockdown of Sidt2 in Cell Lines
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3T3-L1 adipocytes were purchased from the Cell Resource Center of Shanghai Academy of Life Sciences, Chinese Academy of Sciences, while C2-C12 myoblasts and HEPA1-6 hepatoma cells were purchased from ATCC cell bank. 3T3-L1 adipocytes, C2-C12 myoblasts, and HEPA1-6 hepatoma cells were cultured in a DMEM medium containing 10%FBS in an incubator at 37°C and 5% CO2. When the cells grew to the logarithmic phase and the density was 80%, they could be used in the experiment. The above three kinds of cells were transferred to a six-well plate and cultured to about 60% of the cell density, and the lentivirus prepared in advance for knocking out sidt2 and having puro resistance was added to the culture medium. In order to make the lentivirus better-transfected cells, the cells were not treated within 48 hours, and the culture medium could be replaced after 48 hours. 72 hours after the virus was added, the new medium was replaced, and an appropriate amount of puro was added to it to screen cells (the amount of puros varied according to different cell lines). At the same time, it was necessary to add polybrene, to increase the transfection efficiency of the virus. The cells were screened by adding puro for three consecutive days, and the screened cells were cultured normally.
3
Cell Culture and Differentiation Protocol
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HEK293T cells and C2C12 myoblasts (ATCC, New York, NY, USA) were cultured in a growth medium with high-glucose DMEM (Giboc) containing 10% fetal bovine serum (Giboc) in a cell incubator with a humid environment of 37 °C and 5% CO2. The C2C12 cells of between 10 and 25 generations were stimulated with DMEM containing 2% horse serum (Giboc) for differentiation when the degree of cell fusion reached 80%.
4
Immortalized Brown and White Adipogenesis
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Primary brown preadipocytes were isolated from BAT of newborn WT, LSL-K36M;Cre-ER or A-K36M mice. Cells were immortalized by infecting with retroviruses expressing SV40T13 (link). 3T3-L1 cells were from Daniel Lane. Adipogenesis of immortalized brown preadipocytes and the 3T3-L1 white preadipocyte cell line were induced by adding induction medium (Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 0.02 μM insulin, 1 nM T3, 0.5 mM isobutylmethylxanthine, 2 μg ml−1 dexamethasone, and 0.125 mM indomethacin)12 (link). After 2 days of induction, the culture medium was changed to DMEM supplemented with FBS, insulin, and T3 only. PPARγ-stimulated or C/EBPα-stimulated adipogenesis was done by infecting cells with retroviruses expressing PPARγ or C/EBPα before induction33 (link).
C2C12 myoblasts were purchased from ATCC and cultured in growth medium (DMEM supplemented with 15% FBS). Myogenesis was induced by changing growth medium to differentiation medium (DMEM supplemented with 2% horse serum) when cells were confluent8 (link).
C2C12 myoblasts were purchased from ATCC and cultured in growth medium (DMEM supplemented with 15% FBS). Myogenesis was induced by changing growth medium to differentiation medium (DMEM supplemented with 2% horse serum) when cells were confluent8 (link).
5
C2C12 Myoblast Differentiation Protocol
6
Differentiation of C2C12 Myoblasts to Myotubes
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C2C12 myoblasts (2 × 103 cm2, passage number 6; ATCC, Manassas, VA, USA) were cultured in 25 cm2 culture flasks with Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Oud-Beijerland, Holland) supplemented with Glutamax-I (4 mM L-alanyl-L-glutamine), 4.5 g/L glucose (Invitrogen, Carlsbad, CA, USA), and 10% heat-inactivated fetal bovine serum (FBS; Hy-Clone, Oud-Beijerland, Holland). The cells were incubated at 37 °C with 5% CO2 in a humidified atmosphere. Cells were split 1:6 twice weekly and fed 24 h before each experiment. Differentiation into myotubes was achieved via culturing pre-confluent cells (85% confluency) in a medium containing 2% FBS, and they were monitored via microscopy, and for myogenin and MHC expression, via Western blot analysis [74 (link),75 (link)].
7
C2C12 Myoblast Differentiation Protocol
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C2C12 myoblasts (ATCC, Manassas, VA, USA), a mouse myogenic progenitor cell line, were grown in a growth medium (GM; 10% fetal bovine serum (FBS)-containing Dulbecco’s modified Eagle’s medium (DMEM) supplemented with antibiotics (penicillin-streptomycin, 1%, Gibco, Carlsbad, CA, USA) at 37 °C in a 5% CO2 humidified condition. The myoblasts were then cultured on 35 mm plates at a density of 1.3 × 105 in 2 mL of GM. When confluency reached 80 to 90%, a differentiation medium (DM) composed of horse serum (2%, dialyzed, Gibco) in DMEM supplemented with 1% penicillin-streptomycin was used to induce cell differentiation. The medium was refreshed every 24 h. When necessary, cells were pretreated with either bovine serum albumin (BSA) or BSA-conjugated PA (100 μM) in GM for 24 h before differentiation. All reagents and materials were obtained from Sigma-Aldrich unless otherwise specified.
8
Cell Culture Protocols for Cancer Cell Lines
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RD, RH30, HeLa, MCF7, MDA-MB 321, and PC3 cells were obtained from ATCC. RD2, and RH28 were obtained from Dr. Denis Guttridge (Medical University of South Carolina). RD, RD2, RH30, RH28, HeLa, MCF7 and MDA-MB 321 were grown according to standard protocols in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% Fetal Bovine serum (FBS)(Hyclone) and penicillin and streptomycin antibiotics. PC3 cells were grown in RPMI basal media with L-Glutamine supplemented with 10% FBS and penicillin and streptomycin antibiotics. Proliferating C2C12 myoblasts (ATCC) were grown in DMEM supplemented with 10% fetal bovine serum (Hyclone). All cells were grown at 37 oC in a CO2 incubator at 5% CO2. Bio-Synthesis authenticated all RMS cell lines (Lewisville, TX) using STR analysis on September 14, 2011.
9
C2C12 Myoblast Differentiation on IGF-1 Scaffolds
10
Culturing C2C12 and Porcine Satellite Cells
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C2C12 myoblasts were purchased from ATCC (Shanghai, China) and cultured in the proliferation medium (PM). Porcine satellite cells (PSCs) were isolated from large white pigs, as reported previously [28 (link)], and cultured on flasks or plates coated with Matrigel (Corning Inc., Corning, NY, USA) in PM. PM was Dulbecco modified Eagle’s medium (DMEM; Thermo Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Thermo Scientific, Waltham, MA, USA) and 1% penicillin-streptomycin (P/S; Thermo Scientific, Waltham, MA, USA). Cells were passaged every 2–3 days to maintain the confluency < 60%. PSCs at passage 4 (P4) to passage (P8) were used for experiments.
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