KC-5669

293T-cyno-CD79b-Flag Cell Line

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Background of 293T-cyno-CD79b-Flag 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-5669
Cell Line Name	293T-cyno-CD79b-Flag Cell Line
Clone Number	5#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous cyno CD79b Flag 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 + 150ug/mL Hygromycin B
Selection Marker	Hygromycin B
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

Cell Line Generation

293T cyno CD79b Flag cell line was generated using lentiviral vector expressing cyno CD79b Flag sequence.

Characterization

Figure 1: Characterization of cyno CD79b Flag overexpression in the 293T cyno CD79b Flag stable clone using FACS.

Figure 2: Characterization of cyno CD79b Flag expression in 293T cyno CD79b Flag stable clones using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS + 150ug/mL Hygromycin B)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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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.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.
2. 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.