KC-5250

Ramos-CD79B-KO Cell Line

55202

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Background of Ramos-CD79B-KO Cell Line

The CD79B gene encodes the beta subunit of the B cell antigen receptor (BCR) complex, which is crucial for B cell development and immune response. CD79B, along with CD79A, forms the signaling component of the BCR, facilitating antigen recognition and downstream signaling pathways such as those involving SYK and BTK. Mutations in CD79B are implicated in B cell malignancies, including diffuse large B cell lymphoma (DLBCL), where they contribute to constitutive BCR activation and cell survival. CD79B is also a therapeutic target; for example, the antibody drug conjugate polatuzumab vedotin targets CD79B in DLBCL. Studies highlight its role in immune evasion and potential as a biomarker.

Specifications

Catalog Number	KC-5250
Cell Line Name	Ramos-CD79B-KO Cell Line
Clone Number	3A1
Host Cell Line	Ramos
Description	Stable Ramos clone with CD79B gene knockout, No.3A1
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	NA
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:8 every 1-3 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

Ramos-CD79B-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Ramos-CD79B-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of Ramos-CD79B-KO cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of Ramos-CD79B-KO cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS) 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 1:4-1:8 every 1-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 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

Tkachenko A, Kupcova K, Havranek O. B-Cell Receptor Signaling and Beyond: The Role of Igα (CD79a)/Igβ (CD79b) in Normal and Malignant B Cells. Int J Mol Sci. 2023 Dec 19;25(1):10. doi: 10.3390/ijms25010010. PMID: 38203179; PMCID: PMC10779339.
Xu PP, Shen R, Shi ZY, Cheng S, Wang L, Liu Y, Zhang L, Huang R, Ma X, Wu X, Yao H, Yu Y, Zhao WL. The Prognostic Significance of CD79B Mutation in Diffuse Large B-Cell Lymphoma: A Meta-analysis and Systematic Literature Review. Clin Lymphoma Myeloma Leuk. 2022 Dec;22(12):e1051-e1058.e1. doi: 10.1016/j.clml.2022.08.006. Epub 2022 Aug 20. PMID: 36182550.
https://www.ncbi.nlm.nih.gov/gene/974