KC-2111

HEK293T-ARV7-Cell-Line

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Home » 细胞系 » HEK293T-ARV7-Cell-Line

Background of HEK293T-ARV7-Cell-Line

ARv7, also known as AR3. As the most common AR-V, the androgen receptor splice variant 7 (AR-V7), a splice variant of the androgen receptor mRNA resulting in the truncation of the ligand-binding domain, is a promising predictive tumor marker and a relevant selection marker for the treatment of advanced prostate cancer.

Specifications

Catalog NumberKC-2111
Cell Line NameHEK293T-ARV7-Cell-Line
Host Cell LineHuman HEK293T cell line
DescriptionStable HEK293 cell line expressing exogenous human ARv7 gene
QuantityOne vial of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% DMEM+20% FBS+10% DMSO
Propagation MediumDMEM+10%FBS+1μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

HEK293 human ARv7 cell line was generated using a lentiviral vector expressing the human ARv7 sequence.

Characterization

Cell Resuscitation

  1. Prewarm culture medium (DMEM + 10% FBS + 1μg/mL Puromycin) 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 1:3-1:6 every 3 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. The impact of the androgen receptor splice variant AR-V7 on the prognosis and treatment of advanced prostate cancer].2020 Dec;51(6):582-592.
  2. Ligand-independent androgen receptor variants derived from splicing of cryptic exons signify hormonerefractory prostate cancer.2009Jan1;69(1):16-22.
  3. The androgen receptor gene and its influence on the development and progression of prostate cancer.2001 Sep;195(2):138-46.

Use License Agreement

Research Use Only.
Not for use in diagnostic procedures or therapeutic applications.
Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.
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