Cells were isolated from the pectoralis major muscle of a 41-year-old male Caucasian heart-transplant donor, and infected with retrovirus carrying hTERT in pBabeHygro vector to establish the Immortalized Human Skeletal Myoblasts (LHCN-M2). The cells may be used to study study muscular dystrophie, and be used to discover therapy for a variety of age-related pathologies.
Immortalization Method: Immortalized with retrovirus containing hTERT.
BioSafety Level: II
Organism: Human (H. sapiens)
Species: Human
Source Organs: Skeletal Muscle
Organ Type: Skeletal Muscle
Growth Properties: Adherent
Morphology: Polyglonal
Markers: PAX7, desmin, CD56
Donor Age: 41
Donor Gender: Male
Propagation: Gelatin from porcine skin is required (Sigma-Aldrich, G1890).
The base medium for this cell line is a 4 and 1 parts mixture of Prigrow III medium available at abm, Cat. No. TM003, and PriGrow I medium available at abm, Cat. No. TM001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 15%, 1% L-glutamine, 20 mM HEPES, 0.03 µg/ml Zinc sulfate (Sigma-Aldrich, Z0251), 1.4 µg/ml Vitamin B12 (Sigma-Aldrich, V2876), 0.055 µg/ml Dexamethasone (Sigma-Aldrich, D4902), 2.5 ng/ml HGF, 10 ng/ml bFGF, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: +37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.
Quality Control: 1) Immunocytochemistry
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C
Reference: Zhu, C.-H., Mouly, V., Cooper, R. N., Mamchaoui, K., Bigot, A., Shay, J. W., … Wright, W. E. (2007). Cellular senescence in human myoblasts is overcome by human telomerase reverse transcriptase and cyclin-dependent kinase 4: consequences in aging muscle and therapeutic strategies for muscular dystrophies. Aging Cell, 6(4), 515–523. https://doi.org/10.1111/j.1474-9726.2007.00306.x