KC-6433

CHOK1-FASL Cell Line

62432

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Background of CHOK1-FASL Cell Line

FASLG belongs to the tumor necrosis factor (TNF) family and is encoded by a gene located on chromosome 1q23 in human (NCBI Gene ID:356); it is normally stored in the cytoplasm in secretory vesicles and can be expressed as two different isoforms: a 37 kDa membrane-bound protein (mFASLG) and a 26–29 kDa soluble variant (sFASLG), produced by a metalloproteinase-mediated cleavage of the mFASLG. The FASLG gene is composed of four exons and is located on chromosome 1 both in human and mice. It encodes a type II protein, meaning that the N-terminal domain is intracellular.

Specifications

Catalog Number	KC-6433
Cell Line Name	CHOK1-FASL Cell Line
NCBI/UniProt Accession Number	NM_000639
Clone Number	2#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous FASL 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	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:4-1:5 every 2-3 days; seed out at about 1-3 x 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-FASL cell line was generated using a lentiviral vector expressing the FASL sequence.

Characterization

Figure 1: Characterization of FASL overexpression in the CHOK1-FASL stable clone using PCR sequencing.

Figure 2: Characterization of FASL overexpression in the CHOK1-FASL stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS+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 1:4-1:5 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

Magerus A, Bercher-Brayer C, Rieux-Laucat F. The genetic landscape of the FAS pathway deficiencies. Biomed J. 2021 Aug;44(4):388-399. doi: 10.1016/j.bj.2021.06.005. Epub 2021 Jun 24. PMID: 34171534; PMCID: PMC8514852.

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.