KC-3389

CHOK1-cyno-CD79b-Cell-Line

34366

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Background of CHOK1-cyno-CD79b-Cell-Line

CD79b, The B lymphocyte antigen receptor is a multimeric complex that includes the antigen-specific component, surface immunoglobulin (Ig). Surface Ig non-covalently associates with two other proteins, Ig-alpha and Ig-beta, which are necessary for expression and function of the B-cell antigen receptor. This gene encodes the Ig-beta protein of the B-cell antigen component.

Specifications

Catalog Number	KC-3389
Cell Line Name	CHOK1-cyno-CD79b-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHO-K1 cell line expressing exogenous cyno-CD79b 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 1:4-1:5 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

CHOK1 cyno CD79b cell line was generated using a lentiviral vector expressing the cyno CD79b sequence.

Characterization

Figure1: Characterization of CD79b overexpression in CHOK1 cyno-CD79b stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 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

Yamaguchi J, Ohka F, Lushun C, Motomura K, Aoki K, Takeuchi K, Nagata Y, Ito S, Mizutani N, Ohno M, Suzaki N,
Takasu S, Seki Y, Kano T, Wakabayashi K, Oyama H, Kurahashi S, Tanahashi K, Hirano M, Shimizu H, Kitano Y,
Maeda S, Yamazaki S, Wakabayashi T, Kondo Y, Natsume A, Saito R. CD79B Y196 mutation is a potent predictive
marker for favorable response to R-MPV in primary central nervous system lymphoma. Cancer Med. 2023
Mar;12(6):7116-7126. doi: 10.1002/cam4.5512. Epub 2022 Dec 7. PMID: 36478416; PMCID: PMC10067082.
Zheng WW, Zhou H, Li P, Ye SG, Abudureheman T, Yang LT, Qing K, Liang AB, Chen KM, Duan CW. Anti-CD79b/CD3
bispecific antibody combined with CAR19-T cells for B-cell lymphoma treatment. Cancer Immunol Immunother.
2023 Sep 14. doi: 10.1007/s00262-023-03526-z. Epub ahead of print. PMID: 37707586.