KC-5056

CHOK1-mouse-TNFRSF13B-High cell line

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Background of CHOK1-mouse-TNFRSF13B-High cell line

The protein encoded by the TNFRSF13B gene is a lymphocyte-specific member of the tumor necrosis factor (TNF) receptor superfamily. It interacts with calcium modulators and holoprinoprotein ligands (CAML). This protein induces the activation of transcription factors NFAT, AP1, and NF-kappa-B, and plays a crucial role in humoral immunity through its interaction with TNF ligands. This gene is located in the Smith-Magenis syndrome region on chromosome 17.

Specifications

Catalog Number	KC-5056
Cell Line Name	CHOK1-mouse-TNFRSF13B-High cell line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse TNFRSF13B High 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1 mouse TNFRSF13B High cell line was generated using lentiviral vector expressing mouse TNFRSF13B High sequence

Characterization

Figure 1: Characterization of mouse TNFRSF13B High overexpression in the CHOK1 mouse TNFRSF13B High stable clone using FACS.

Figure 2: Characterization of mouse TNFRSF13B High overexpression in the CHOK1 mouse TNFRSF13B High stable clone using PCR sequence

Cell Resuscitation

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

Cell Freezing

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

References

1. Pulvirenti F, Zuntini R, Milito C, Specchia F, Spadaro G, Danieli MG, Pession A, Quinti I, Ferrari S. Clinical Associations of Biallelic and Monoallelic TNFRSF13B Variants in Italian Primary Antibody Deficiency Syndromes. J Immunol Res. 2016;2016:8390356. doi: 10.1155/2016/8390356. Epub 2016 Mar 30. PMID: 27123465; PMCID: PMC4829724.
2.Cascalho M, Platt JL. TNFRSF13B in B cell responses to organ transplantation. Hum Immunol. 2023 Jan;84(1):27-33. doi: 10.1016/j.humimm.2022.09.006. Epub 2022 Nov 1. PMID: 36333165; PMCID: PMC10429825.
3.Li CY, Huang SP, Chen YT, Wu HE, Cheng WC, Huang CY, Yu CC, Lin VC, Geng JH, Lu TL, Bao BY. TNFRSF13B is a potential contributor to prostate cancer. Cancer Cell Int. 2022 May 6;22(1):180. doi: 10.1186/s12935-022-02590-2. PMID: 35524261; PMCID: PMC9074181.