Cas9 Expressing Immortalized Human Skeletal Muscle Cell Line

Immortalized human skeletal muscle cells stably expressing Cas9

Organism: Human

Source Organ: Skeletal Muscle

BioSafety Level: II

Quantity: 10⁶ cells/1.0 ml

Growth Properties: Adherent

Morphology: Multipolar

Markers: Neomycin

Propagation: The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the completed growth medium, add the following components to the base medium with the final concentration: fetal bovine serum (TM999) to a final concentration of 10%, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.

Freeze Thaw: 1. Pre-warm complete media in a +37°C waterbath. 2. Remove the cryopreserved vial from the liquid nitrogen storage tank. 3. Thaw the cells quickly by placing the lower half of the vial into the +37°C water bath and remove after 60 seconds. There should still be a few ice crystals left after thawing. It is important not to over-thaw the cryovials as the presence of DMSO is toxic to the cells. 4. Resuspend the cells in the vial and transfer the cell suspension into a 15 ml sterile conical tube containing 5 ml complete media. 5. Centrifuge cells at 200x g for 3 minutes to pellet. 6. Aspirate out the media, leaving cell pellet undisturbed. 7. Resuspend pellet in fresh culture medium and plate in new culture vessel. 8. Incubate cultures at +37°C, 5% CO₂.

Quality Control: 1) Neomycin drug selection

2) Western Blot analysis for Cas9 transgene expression

Shipping Conditions: Dry Ice

Storage Conditions: -180°C