KC-3714

CHOK1-EPHA1 Cell Line

34648

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Background of CHOK1-EPHA1 Cell Line

EPHA1 (EPH Receptor A1) also known as EPH, it is a Protein Coding gene. EphA1 Protein, a receptor tyrosine kinase, engages in bidirectional signaling with membrane-bound ephrin-A ligands. Activation by EFNA1 regulates cell attachment, motility, angiogenesis, and proliferation. EphA1 may also play a role in apoptosis. Diseases associated with EPHA1 include Craniofrontonasal Syndrome and Retinitis Pigmentosa 18. Among its related pathways are GPCR Pathway and ERK Signaling.

Specifications

Catalog Number	KC-3714
Cell Line Name	CHOK1-EPHA1 Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human EPHA1 gene
Quantity	One vial 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-EPHA1 Cell Line was generated using a lentiviral vector expressing the human EPHA1 equence.

Characterization

Figure 1: Characterization of human EPHA1 overexpression in the CHO-K1 EPHA1 stable clone using FACS.

Figure 2: Characterization of human EPHA1 and its mutants overexpressing in CHOK1 stable clones 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. Ieguchi K, Maru Y. Roles of EphA1/A2 and ephrin-A1 in cancer. Cancer Sci. 2019 Mar;110(3):841-848. doi: 10.1111/cas.13942. Epub 2019 Feb 15. PMID: 30657619; PMCID: PMC6398892. 2. Wu Y, Du Z, Mou J, Qiu X, Chen J, Cai S, Ren D, Xiao F, Zhou G, Yuan C. The Functions of EphA1 Receptor Tyrosine Kinase in Several Tumors. Curr Med Chem. 2023;30(20):2340-2353. doi: 10.2174/0929867329666220820125638. PMID: 35996244. 2. Ieguchi K, Maru Y. Roles of EphA1/A2 and ephrin-A1 in cancer. Cancer Sci. 2019 Mar;110(3):841-848. doi: 10.1111/cas.13942. Epub 2019 Feb 15. PMID: 30657619; PMCID: PMC6398892. 4. Pasquale EB. Eph receptors and ephrins in cancer: bidirectional signalling and beyond. Nat Rev Cancer. 2010 Mar;10(3):165-80. doi: 10.1038/nrc2806. PMID: 20179713; PMCID: PMC2921274.

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.