| Field | Specification |
|---|---|
| Species |
CaSki is a cell line exhibiting epithelial morphology, isolated from the cervix of a 40-year-old White female patient with epidermoid carcinoma. The establishment of this cell line provides a critical model for the study of cervical cancer, particularly in the context of HPV-mediated oncogenesis. CaSki cells are characterized by their capacity to replicate HPV16 DNA, which is integrated into the host's genome, offering insights into the viral life cycle and its role in malignant transformation.
These cells are an essential resource in cancer research, particularly for studies focusing on the pathogenesis of HPV-associated cervical cancer. The presence of high-risk HPV16 in CaSki cells facilitates the exploration of viral oncogene functions, notably the E6 and E7 proteins and their interactions with cellular tumor suppressor pathways, including those involving p53 and pRB. This aspect makes CaSki cells invaluable for evaluating potential therapeutic targets and developing interventions aimed at HPV-induced malignancies.
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- Isoenzymes: G6PD, B
- Products: beta subunit of hCG, tumor associated antigen
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2 will result in a confluent monolayer within 3to4 days.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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