KC-4573

CHOK1-mouse-ITGAV Cell Line

47745

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Background of CHOK1-mouse-ITGAV Cell Line

ITGAV (Integrin Subunit Alpha V) also known as CD51, it is a Protein Coding gene. Integrins are heterodimeric integrated membrane proteins composed of alpha and beta subunits, which play a role in cell surface adhesion and signal transduction. This subunit binds to β 1, β 3, β 5, β 6, and β 8 subunits. The heterodimer composed of α V and β 3 subunits is also known as a fibronectin receptor. This integrin may regulate angiogenesis and cancer progression.Diseases associated with ITGAV include Visceral Myopathy 1 and West Nile Virus.

Specifications

Catalog Number	KC-4573
Cell Line Name	CHOK1-mouse-ITGAV Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse ITGAV 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

Cell Line Generation

CHOK1-mouse-ITGAV Cell Line was generated using a lentiviral vector expressing the mouse ITGAV sequence.

Characterization

Figure 1: Characterization of mouse ITGAV overexpression in the CHO-K1 mouse ITGAV stable clone using QPCR.

Figure 2: Characterization of mouse ITGAV overexpression in the CHO-K1 mouse ITGAV stable clone using PCR sequence.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/mL Puromycin)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Loeser, H., Scholz, M., Fuchs, H. et al. Integrin alpha V (ITGAV) expression in esophageal adenocarcinoma is associated with shortened overall-survival. Sci Rep 10, 18411 (2020).
Cheuk IW, Siu MT, Ho JC, Chen J, Shin VY, Kwong A. ITGAV targeting as a therapeutic approach for treatment of metastatic breast cancer. Am J Cancer Res. 2020 Jan 1;10(1):211-223. PMID: 32064162; PMCID: PMC7017729.
Frank JW, Seo H, Burghardt RC, Bayless KJ, Johnson GA. ITGAV (alpha v integrins) bind SPP1 (osteopontin) to support trophoblast cell adhesion. Reproduction. 2017 May;153(5):695-706. doi: 10.1530/REP-17-0043. Epub 2017 Mar 1. PMID: 28250242.