KC-0792

VCAP-Cell-Line-(Not-for-sale)

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Background of VCAP-Cell-Line-(Not-for-sale)

VCAP was established in 1997 from a vertebral bone metastasis from a patient with hormone refractory prostate cancer.

Specifications

Catalog NumberKC-0792
Cell Line NameVCAP-Cell-Line-(Not-for-sale)
Host Cell LineNA
DescriptionDerived from a vertebral metastasis of a human prostate carcinoma.
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
Application3D cell culture; Cancer research
Freezing Medium70% DMEM+20% FBS+10% DMSO
Propagation MediumDMEM+10%FBS
Selection MarkerNA
MorphologyEpithelial
SubcultureSplit saturated culture 1:2-1:3 every 3-4 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 53-144 hours
Mycoplasma StatusNegative
In Vivo ValidationYes

Cell Line Generation

NA

Characterization

Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:2-1:3 every 3-4 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1. Parallel genome-scale loss of function screens in 216 cancer cell lines for the identification of context-specific genetic dependencies. Sci. Data 1:140035.1-140035.12(2014)
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