KC-4801

CHOK1-OX40L Cell Line

55558

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Background of CHOK1-OX40L Cell Line

OX40L, as known as TNFSF4. This gene encodes a cytokine of the tumor necrosis factor (TNF) ligand family. The encoded protein functions in T cell antigen-presenting cell (APC) interactions and mediates adhesion of activated T cells to endothelial cells. Polymorphisms in this gene have been associated with Sjogren\'s syndrome and systemic lupus erythematosus. Alternative splicing results in multiple transcript variants.

Specifications

Catalog Number	KC-4801
Cell Line Name	CHOK1-OX40L Cell Line
Clone Number	1-6#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous OX40L 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1 OX40L cell line was generated using a lentiviral vector expressing the OX40L sequence.

Characterization

Figure 1: Characterization of OX40L overexpression in the CHOK1 OX40L stable clone using FACS.

Figure 2: Characterization of OX40L and its mutants overexpressing in CHOK1 stable clone using PCR sequencing.

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

Kondo K, Okuma K, Tanaka R, Matsuzaki G, Ansari AA, Tanaka Y. Rapid induction of OX40 ligand on primary T cells activated under DNA-damaging conditions. Hum Immunol. 2008 Sep;69(9):533-42. doi: 10.1016/j.humimm.2008.07.001. Epub 2008 Aug 15. PMID: 18718855.
Olofsson PS, Söderström LA, Jern C, Sirsjö A, Ria M, Sundler E, de Faire U, Wiklund PG, Ohrvik J, Hedin U, Paulsson-Berne G, Hamsten A, Eriksson P, Hansson GK. Genetic variants of TNFSF4 and risk for carotid artery disease and stroke. J Mol Med (Berl). 2009 Apr;87(4):337-46. doi: 10.1007/s00109-008-0412-5. Epub 2008 Nov 8. PMID: 18998106.

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.