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	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 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.