KC-5132

CHOK1-mouse-BAFFR cell line

55316

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Background of CHOK1-mouse-BAFFR cell line

The B lymphocyte stimulator receptor (BAFFR, also known as TNFRSF13C, BROMIX, CD268, CVID4, prolixin) is encoded by the TNFRSF13C gene and is one of the main membrane protein receptors for B cell proliferation and differentiation. It belongs to the TNF receptor (TNFR) superfamily, but BAFFR differs from other TNFRs in that its CRDs (cysteine-rich domains) contain only four cysteine residues, which are used for ligand binding and self-assembly.

Specifications

Catalog Number	KC-5132
Cell Line Name	CHOK1-mouse-BAFFR cell line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse BAFFR 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 mouse-BAFFR cell line was generated using lentiviral vector expressing mouse-BAFFR sequence

Characterization

Figure 1: Characterization of mouse-BAFFR overexpression in the CHOK1 mouse-BAFFR stable clone using FACS.

Figure 2: Characterization of mouse-BAFFR overexpression in the CHOK1 mouse-BAFFR 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. Ntellas P, Dardiotis E, Sevdali E, Siokas V, Aloizou AM, Tsinti G, Germenis AE, Hadjigeorgiou GM, Eibel H, Speletas M. TNFRSF13C/BAFFR P21R and H159Y polymorphisms in multiple sclerosis. Mult Scler Relat Disord. 2020 Jan;37:101422. doi: 10.1016/j.msard.2019.101422. Epub 2019 Sep 30. PMID: 32172995.
2.Prieto-Peña D, Genre F, Remuzgo-Martínez S, Pulito-Cueto V, Atienza-Mateo B, Llorca J, Sevilla-Pérez B, Ortego-Centeno N, Lera-Gómez L, Leonardo MT, Peñalba A, Narváez J, Martín-Penagos L, Rodrigo E, Miranda-Filloy JA, Caminal-Montero L, Collado P, Pérez JS, de Argila D, Rubio E, Luque ML, Blanco-Madrigal JM, Galíndez-Agirregoikoa E, Gualillo O, Martín J, Castañeda S, Blanco R, González- Gay MA, López-Mejías R. BAFF, APRIL and BAFFR on the pathogenesis of Immunoglobulin-A vasculitis. Sci Rep. 2021 Jun 1;11(1):11510. doi: 10.1038/s41598-021-91055-z. Erratum in: Sci Rep. 2021 Aug 16;11(1):16946. doi: 10.1038/s41598-021-96114-z. PMID: 34075170; PMCID: PMC8169776.
3.Müller-Winkler J, Mitter R, Rappe JCF, Vanes L, Schweighoffer E, Mohammadi H, Wack A, Tybulewicz VLJ. Critical requirement for BCR, BAFF, and BAFFR in memory B cell survival. J Exp Med. 2021 Feb 1;218(2):e20191393. doi: 10.1084/jem.20191393. PMID: 33119032; PMCID: PMC7604764.

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.