KC-3015

293T-FIBCD1-FGL2 Cell Line

34062

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Background of 293T-FIBCD1-FGL2 Cell Line

FIBCD1 is a conserved type II transmembrane endocytic receptor that binds chitin and is located primarily in the intestinal brush border.
FGL2, also known as Fibrinogen Like 2. The protein encoded by this gene is a secreted protein that is similar to the beta- and gamma-chains of fibrinogen. The carboxyl-terminus of the encoded protein consists of the fibrinogen-related domains (FRED). The encoded protein forms a tetrameric complex which is stabilized by interchain disulfide bonds. This protein may play a role in physiologic functions at mucosal sites.

Specifications

Catalog Number	KC-3015
Cell Line Name	293T-FIBCD1-FGL2 Cell Line
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous FIBCD1-FGL2 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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
In Vivo Validation	NA

Cell Line Generation

293T-FIBCD1-FGL2 cell line was generated using a lentiviral vector expressing the FIBCD1-FGL2 sequence.

Characterization

Figure 1: Characterization of FIBCD1-FGL2 overexpression in the 293T-FIBCD1-FGL2 stable clone using FACS.(Primary antibody: 9D8-hIgG1, Cat#KB-1151, Kyinno)

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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% DMEM + 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

Schlosser A, Thomsen T, Moeller JB, Nielsen O, Tornøe I, Mollenhauer J, Moestrup SK, Holmskov U. Characterization of FIBCD1 as an acetyl group-binding receptor that binds chitin. J Immunol. 2009 Sep 15;183(6):3800-9. doi: 10.4049/jimmunol.0901526. Epub 2009 Aug 26. PMID: 19710473.
Ma X, Zhu H, Cheng L, Chen X, Shu K, Zhang S. Targeting FGL2 in glioma immunosuppression and malignant progression. Front Oncol. 2022 Oct 13;12:1004700. doi: 10.3389/fonc.2022.1004700. PMID: 36313679; PMCID: PMC9606621.
Zhao Q, Hu J, Kong L, Jiang S, Tian X, Wang J, Hashizume R, Jia Z, Fowlkes NW, Yan J, Xia X, Yi SF, Dao LH, Masopust D, Heimberger AB, Li S. FGL2-targeting T cells exhibit antitumor effects on glioblastoma and recruit tumor-specific brain-resident memory T cells. Nat Commun. 2023 Feb 10;14(1):735. doi: 10.1038/s41467-023-36430-2. PMID: 36759517; PMCID: PMC9911733.