KC-2114

CHOK1-PLAUR-Cell-Line

23418

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Background of CHOK1-PLAUR-Cell-Line

PLAUR (Plasminogen Activator, Urokinase Receptor) also known as UPAR, This gene encodes the receptor for urokinase plasminogen activator, Urokinase-type plasminogen activator receptor (uPAR) is a glycolipid-anchored protein on the cell surface, is controlled by many factors in tumorigenesis and is expressed in many tumor tissues. The role of PLAUR in targeting and promoting plasminogen formation, may influence many normal and pathological processes associated with cell surface plasminogen activation and local degradation of extracellular matrix.

Specifications

Catalog Number	KC-2114
Cell Line Name	CHOK1-PLAUR-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous PLAUR 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 PLAUR Cell Line was generated using a lentiviral vector expressing the PLAUR sequence.

Characterization

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

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

Ballonová L, Kulíšková P, Slanina P, Štíchová J, Vlková M, Hakl R, Litzman J, Souček P, Freiberger T. PLAUR splicing pattern in hereditary angioedema patients' monocytes and macrophages. Mol Biol Rep. 2023 Jun;50(6):4975-4982. doi: 10.1007/s11033-023-08391-8. Epub 2023 Apr 22. PMID: 37086298.
Li J, Fan H, Zhou X, Xiang Y, Liu Y. Prognostic Significance and Gene Co-Expression Network of PLAU and PLAUR in Gliomas. Front Oncol. 2022 Jan 11;11:602321. doi: 10.3389/fonc.2021.602321. PMID: 35087738; PMCID: PMC8787124.
Lv T, Zhao Y, Jiang X, Yuan H, Wang H, Cui X, Xu J, Zhao J, Wang J. uPAR: An Essential Factor for Tumor Development. J Cancer. 2021 Oct 17;12(23):7026-7040. doi: 10.7150/jca.62281. PMID: 34729105; PMCID: PMC8558663.