C2C12 Cell Line

Brief Introdution
Product List

General Information
Organism	Mus musculus, mouse
Cell Line Description	C2C12 is an immortalized mouse myoblast cell line. The C2C12 cell line is a subclone of myoblasts originally obtained by Yaffe and Saxel in 1977 at the Weizmann Institute of Science, Israel. C2C12 cells exhibit rapid development and maturation into functional skeletal muscle cells or cardiomyocytes with the ability to contract and generate force. The rate of muscle formation in C2C12 cells can be controlled by introducing loss-of-function genes critical for myoblast fusion and myogenesis. Under necrotic conditions, such as tumor necrosis factor alpha (TNF-α), there is a direct loss of proteins in C2C12 skeletal muscle cells, particularly myosin heavy chain proteins. C2C12 cells were used to elucidate X chromosome (Xi) replication that is inactive during early S phase of the cell cycle and is subject to epigenetic regulation. C2C12 cells are particularly convenient for studying the cell cycle due to their high division rate. C2C12 cells are used to study the differentiation and myogenesis of myoblasts and osteoblasts, express various target proteins, and explore mechanistic biochemical pathways.
Tissue	Muscle
Disease	Normal
Morphology	Myoblast
Gender	Female
Age	Adult
Product Format	2 months
Growth Mode	Adherent
Biosafety Level	1 (2 when used for genetic engineering) Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Applications	1. Study of muscle differentiation 2. Drug testing 3. Genetic studies 4. Study of cellular processes 5. Disease modelling 6. Protein studies
Shipped In	Dry ice
Storage Temperature	−196°C
Additional Info	C2C12 cells have been shown to efficiently incorporate exogenous cDNA and nucleic acids via transfection. In an initial pilot study by Yaffe and Saxel, C2C12 was obtained by serial passage of myoblasts cultured from thigh muscles of C3H mice after crush injury. In their study, a group of C2C12 cells were cultured from normal mouse myoblasts cultured after crush injury in two-month-old C3H mice. Within two days, normal cells differentiate into spindle-shaped mononucleated myoblasts. After four days, a multinucleated myotube network forms, and after a few days, sarcomeres and Z-lines can be observed. Instead, dystrophic cells form shortened fibers that are covered by fibroblasts, a hallmark of muscle atrophy.
Culture Conditions and Handling
Subculturing	1. Remove old culture medium from adherent cells and wash with calcium- and magnesium-free PBS. For T25 flasks, use 3-5 ml of PBS and for T75 flasks, use 5-10 ml. 2. Then, completely cover the cells with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. 3. Allow cells to detach by incubating at room temperature for 8-10 minutes. 4. After incubation, resuspend cells by gently mixing with 10 ml of medium and centrifuge at 300xg for 3 minutes. 5. Discard the supernatant, resuspend the cells in fresh medium, and transfer them to a new flask that already contains fresh medium.
Medium Renewal	Every 3 to 5 days
Subcultivation Ratio	A split ratio of 1:3 to 1:5 is recommended.
Culture Medium	DMEM + 2mM Glutamine + 10-15% Foetal Bovine Serum FBS / FCS.
Culture Conditions	Atmosphere: Air, 95%; CO₂, 5%;Temperature: 37°C
Cryopreservation	Complete growth medium supplemented with 5% (v/v) DMSO

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* For research use only. Not intended for any clinical use.

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