Ovcar3 cells stably expressing Cas9Organism: HumanSource Organ: OvaryBioSafety Level: IIQuantity: 10⁶ cells/1.0 mlGrowth Properties: AdherentMorphology: Epithelial-LikeMarkers: PuromycinPropagation: The base medium for this cell line is Prigrow II medium (TM002). To make the completed growth medium, add the following components to the base medium with the final concentration: Heat-Inactivated fetal bovine serum to a final concentration of 10%, L-glutamine to a final concentration of 1%, Sodium Pyruvate to a final concentration of 1%, NEAA to a final concentration of 1%, 0.5 µg/ml puromycin (G264) 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) Puromycin selection2) Western Blot analysis for Cas9 transgene expressionShipping Conditions: Dry IceStorage Conditions: -180°C