KC-5384

CHOK1-CDH20 Cell Line

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Home » CHOK1-CDH20 Cell Line

Background of CHOK1-CDH20 Cell Line

Cancer-associated cadherin 20 (CDH20) is a novel identified cadherin that is genetically altered in several types of human cancer, including cervical cancer. However, its involvement in the progression of cervical cancer remains unknown. In this study, we show that CDH20 was downregulated in clinical cervical cancer samples and its expression correlated with cervical cancer clinical features. CDH20 negatively regulated the migration and invasion of cervical cancer cells.

Specifications

Catalog NumberKC-5384
Cell Line NameCHOK1-CDH20 Cell Line
NCBI/UniProt Accession NumberNM_022843.3
Clone Number2#
Host Cell LineCHOK1
DescriptionStable CHOK1 cell line expressing exogenous CDH20 gene
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% basal medium+20% FBS+10% DMSO
Propagation MediumRPMI 1640+10% FBS +10μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial-like
SubcultureSplit saturated culture 1:6-1:8 every 2-3 days
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative

Cell Line Generation

CHOK1-CDH20 cell line was generated using a lentiviral vector expressing the CDH20 sequence.

Characterization

Cell Resuscitation

  1. Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
  2. Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
  3. Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
  4. Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
  5. Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
  6. Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
  7. Incubate the flask in a 37°C in a humidified 5% CO2 incubator.
  8. Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
  9. Subculture the cells at a ratio of 1:6-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

  1. Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
  2. Pre-chill the freezing medium on ice and label the cryovials accordingly.
  3. Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
  4. Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
  5. Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×106 cells/mL.
  6. Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
  7. Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
  8. Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1.Li C, Ao H, Chen G, Wang F, Li F. The Interaction of CDH20 With β-Catenin Inhibits Cervical Cancer Cell Migration and Invasion via TGF-β/Smad/SNAIL Mediated EMT. Front Oncol. 2020 Jan 9;9:1481.
2. Takahashi M, Fukabori R, Kawasaki H, Kobayashi K, Kawakami K. The distribution of Cdh20 mRNA demarcates somatotopic subregions and subpopulations of spiny projection neurons in the rat dorsolateral striatum. J Comp Neurol. 2021 Nov;529(16):3655-3675.

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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