KC-6437

CHOK1-cyno-SLITRK6 Cell Line

×
请在浏览器中启用JavaScript来完成此表单。
63981
Home » CHOK1-cyno-SLITRK6 Cell Line

Background of CHOK1-cyno-SLITRK6 Cell Line

SLITRK6 encodes a member of the SLITRK protein family. Members of this family are integral membrane proteins that are characterized by two N-terminal leucine-rich repeat (LRR) domains and a C-terminal region that shares homology with trk neurotrophin receptors. This protein functions as a regulator of neurite outgrowth required for normal hearing and vision. Mutations in this gene are a cause of myopia and deafness.

Specifications

Catalog NumberKC-6437
Cell Line NameCHOK1-cyno-SLITRK6 Cell Line
NCBI/UniProt Accession NumberXM_005586081.5
Clone Number10#
Host Cell LineCHOK1
DescriptionStable CHOK1 cell line expressing exogenous cyno-SLITRK6 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% basal medium+20% FBS+10% DMSO
Propagation MediumRPMI 1640+10% FBS +10μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial-like
SubcultureSplit saturated culture 1:4-1:8 every 2-3 days
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

CHOK1-cyno-SLITRK6 cell line was generated using a lentiviral vector expressing the cyno-SLITRK6 sequence.

Characterization

Figure 1: Characterization of cyno-SLITRK6 overexpression in the CHOK1-cyno-SLITRK6 stable clone using QPCR.

Figure 2: Characterization of cyno-SLITRK6 in the CHOK1-cyno-SLITRK6 stable clone using PCR sequencing.

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 1:4-1:8 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.Yu F, Zhao X, Li M, Meng M. SLITRK6 promotes the progression of lung adenocarcinoma by regulating PI3K/AKT/mTOR signaling and Warburg effect. Apoptosis. 2023 Aug;28(7-8):1216-1225.
2.Liu X, Liu Y, Liu Z, Zhang Y, Ma Y, Bai J, Yao H, Wang Y, Zhao X, Li R, Song X, Chen Y, Feng Z, Wang L. Identification of SLITRK6 as a Novel Biomarker in hepatocellular carcinoma by comprehensive bioinformatic analysis. Biochem Biophys Rep. 2021 Oct 27;28:101157.

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.
请在浏览器中启用JavaScript来完成此表单。