KC-5231

CHOK1-TNFRSF13B-Low Cell Line

56059

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Background of CHOK1-TNFRSF13B-Low Cell Line

TNFRSF13B (TNF Receptor Superfamily Member 13B) is a Protein Coding gene. The protein encoded by this gene is a lymphocyte-specific member of the tumor necrosis factor (TNF) receptor superfamily. It interacts with calcium-modulator and cyclophilin ligand (CAML). The protein induces activation of the transcription factors NFAT, AP1, and NF-kappa-B and plays a crucial role in humoral immunity by interacting with a TNF ligand. This gene is located within the Smith-Magenis syndrome region on chromosome 17. Diseases associated with TNFRSF13B include Immunodeficiency, Common Variable, 2 and Immunoglobulin A Deficiency 2.

Specifications

Catalog Number	KC-5231
Cell Line Name	CHOK1-TNFRSF13B-Low Cell Line
Clone Number	3-5#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human TNFRSF13B gene in low level
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

Cell Line Generation

CHOK1-TNFRSF13B-Low Cell Line was generated using a lentiviral vector expressing the human TNFRSF13B sequence.

Characterization

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

Figure 2: Characterization of human TNFRSF13B and its mutants overexpressing in CHOK1 stable clones using PCR sequencing.

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. Platt JL, de Mattos Barbosa MG, Huynh D, Lefferts AR, Katta J, Kharas C, Freddolino P, Bassis CM, Wobus C, Geha R, Bram R, Nunez G, Kamada N, Cascalho M. TNFRSF13B polymorphisms counter microbial adaptation to enteric IgA. JCI Insight. 2021 Jul 22;6(14):e148208. doi: 10.1172/jci.insight.148208. PMID: 34111031; PMCID: PMC8410086.
2. Cumbo C, Orsini P, Tarantini F, Anelli L, Zagaria A, Tragni V, Coccaro N, Tota G, Parciante E, Conserva MR, Redavid I, Minervini CF, Minervini A, Attolico I, Gentile M, Pierri CL, Specchia G, Musto P, Albano F. TNFRSF13B gene mutation in familial acute myeloid leukemia: A new piece in the complex scenario of hereditary predisposition? Hematol Oncol. 2023 Dec;41(5):942-946. doi: 10.1002/hon.3212. Epub 2023 Aug 3. PMID: 37534633.
3.Went M, Duran-Lozano L, Halldorsson GH, Gunnell A, Ugidos-Damboriena N, Law P, Ekdahl L, Sud A, Thorleifsson G, Thodberg M, Olafsdottir T, Lamarca-Arrizabalaga A, Cafaro C, Niroula A, Ajore R, Lopez de Lapuente Portilla A, Ali Z, Pertesi M, Goldschmidt H, Stefansdottir L, Kristinsson SY, Stacey SN, Love TJ, Rognvaldsson S, Hajek R, Vodicka P, Pettersson-Kymmer U, Späth F, Schinke C, Van Rhee F, Sulem P, Ferkingstad E, Hjorleifsson Eldjarn G, Mellqvist UH, Jonsdottir I, Morgan G, Sonneveld P, Waage A, Weinhold N, Thomsen H, Försti A, Hansson M, Juul-Vangsted A, Thorsteinsdottir U, Hemminki K, Kaiser M, Rafnar T, Stefansson K, Houlston R, Nilsson B. Deciphering the genetics and mechanisms of predisposition to multiple myeloma. Nat Commun. 2024 Aug 5;15(1):6644. doi: 10.1038/s41467-024-50932-7. PMID: 39103364; PMCID: PMC11300596.