Skeletal muscle differentiation medium

Manufactured by PromoCell

Sourced in United States

Skeletal Muscle Differentiation Medium is a cell culture medium designed to support the differentiation of skeletal muscle cells. It provides the necessary nutrients and growth factors to facilitate the transition of myoblasts into mature, multinucleated myotubes.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Skeletal muscle growth medium, by PromoCell (1 mentions) Blood cell culture dna mini kit, by Qiagen (1 mentions) Jetpei, by Polyplus Transfection (1 mentions) Mesencult acf chondrogenic differentiation kit, by STEMCELL (1 mentions) Rock inhibitor y 27632, by MedChemExpress (1 mentions) 35 mm culture dish, by Corning (1 mentions) Osteomax xf differentiation medium, by Merck Group (1 mentions)

2 protocols using skeletal muscle differentiation medium

Evaluate CRISPR/Cas9 Targeting for DMD

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To preliminarily screen the activity of the candidate gRNAs, we transfected HEK293T cells with the Cas9 plasmid JDS246 (Addgene number 43861) and with each gRNA, using Lipofectamine2000 (Thermo Fisher Scientific). DNA extraction (Blood & Cell Culture DNA Mini Kit, QIAGEN) was performed 72 hr after transfection for PCR amplification and T7E1 assay.
Immortalized DMD myoblasts were maintained in Skeletal Muscle Growth Medium (Promocell) and differentiated into myotubes for at least 7 days in Skeletal Muscle Differentiation Medium (Promocell) at 37°C with 5% CO₂ incubation. Transfection of spCas9 and DMD gRNAs was performed with JetPEI (Polyplus) following manufacturer’s instructions. After 72 hr, DNA was extracted to perform PCR amplification.

Lattanzi A., Duguez S., Moiani A., Izmiryan A., Barbon E., Martin S., Mamchaoui K., Mouly V., Bernardi F., Mavilio F, & Bovolenta M. (2017). Correction of the Exon 2 Duplication in DMD Myoblasts by a Single CRISPR/Cas9 System. Molecular Therapy. Nucleic Acids, 7, 11-19.

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Adipose-Derived Stem Cell Differentiation

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ASCs were seeded at 10.2 × 10⁴ ASC per cm² in a 35-mm culture dish (Corning, Corning, NY). The ASCs were cultured with RoosterNourish™-MSC-XF culture media (RoosterBio, Inc., Ballenger Creek, Maryland). The differentiation of the cell sheets started when they reached confluence, on day 4 after initial seeding.

- Undifferentiated ASCCSs: ASCs were cultured with RoosterNourish™-MSC-XF. The culture media were replaced every 2 days, up to 18 days from the initial seeding day.

- Osteoblast cell sheets: ASCs were cultured with Osteomax-XF Differentiation Medium (Millipore-Sigma, Burlington, MA). The culture media were replaced every 3 days, up to 17 days from the initial seeding day.

- Chondrocyte cell sheets: ASCs were cultured with a MesenCult™-ACF Chondrogenic Differentiation Kit (Stem Cell, Vancouver, Canada). The culture media were replaced every 2 days, up to 19 days from the initial seeding, and were also replaced when 10 M of ROCK Inhibitor y-27632 (MedChemExpress, Monmouth Junction, NJ, United States) was used.

- Skeletal muscle differentiation medium (PromoCell, Heidelberg, Germany) was used to show that the cells do not form a multilayer cell sheet and the cell density does not change by measuring the transmittance.

Ochiai J., Villanueva L., Niihara H., Niihara Y, & Oliva J. (2022). Posology and Serum-/Xeno-Free Engineered Adipose Stromal Cells Cell Sheets. Frontiers in Cell and Developmental Biology, 10, 873603.

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