KC-5285

CHOK1-PTK7 Cell Line

57155

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Background of CHOK1-PTK7 Cell Line

PTK7, also known as protein tyrosine kinase 7 or CCK4, is a non-receptor protein tyrosine kinase encoded by the PTK7 gene. It mainly plays a role in development and immune responses. The mechanism of PTK7 involves interacting with various proteins and participating in processes such as cell polarity, migration, and proliferation, especially in embryonic development and tumor cell invasion. PTK7 is involved in the regulation of multiple intracellular signaling pathways, including the Wnt/PCP and canonical Wnt signaling pathways, influencing cell polarity, movement, migration, and invasion.

Specifications

Catalog Number	KC-5285
Cell Line Name	CHOK1-PTK7 Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human PTK7 gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/ml
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1 human PTK7 Cell Line was generated using a lentiviral vector expressing the human PTK7 sequence.

Characterization

Figure 1: Characterization of PTK7 overexpression in the CHOK1 PTK7 stable clone using FACS.

Figure 2: Characterization of PTK7 overexpression in the CHOK1 PTK7 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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. Damelin M, Bankovich A, Bernstein J, Lucas J, Chen L, Williams S, Park A,Aguilar J, Ernstoff E, Charati M, Dushin R, Aujay M, Lee C, Ramoth H, Milton M,Hampl J, Lazetic S, Pulito V, Rosfjord E, Sun Y, King L, Barletta F, Betts A,Guffroy M, Falahatpisheh H, O'Donnell CJ, Stull R, Pysz M, Escarpe P, Liu D,Foord O, Gerber HP, Sapra P, Dylla SJ. A PTK7-targeted antibody-drug conjugatereduces tumor-initiating cells and induces sustained tumor regressions. SciTransl Med. 2017 Jan 11;9(372):eaag2611. doi: 10.1126/scitranslmed.aag2611.PMID: 28077676.
2.Jin Z, Guo T, Zhang X, Wang X, Liu Y. PTK7: an underestimated contributor tohuman cancer. Front Oncol. 2024 Oct 15;14:1448695. doi:10.3389/fonc.2024.1448695. PMID: 39474113; PMCID: PMC11518688.
3.Frenster JD, Erdjument-Bromage H, Stephan G, Ravn-Boess N, Wang S, Liu W,Bready D, Wilcox J, Kieslich B, Jankovic M, Wilde C, Horn S, Sträter N,Liebscher I, Schöneberg T, Fenyo D, Neubert TA, Placantonakis DG. PTK7 is apositive allosteric modulator of GPR133 signaling in glioblastoma. Cell Rep.2023 Jul 25;42(7):112679. doi: 10.1016/j.celrep.2023.112679. Epub 2023 Jun 23.PMID: 37354459; PMCID: PMC10445595.