Name: CHOK1-CD79b-High-Cell-Line - Kyinno Bio
SKU: KC-2949

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Background of CHOK1-CD79b-High-Cell-Line

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-2949

Cell Line Name:

CHOK1-CD79b-High-Cell-Line

Host Cell Line:

CHOK1

Description:

Stable CHOK1 cell line expressing exogenous CD79b gene at a high level

Quantity:

One vial of frozen cells (5×10⁶ per 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 + 10µg/mL Puromycin

Selection Marker:

Puromycin

Morphology:

Epithelial-like

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

In Vivo Validation:

Cell Line Generation

CHOK1-CD79b-High Cell Line was generated using lentivirus expressing CD79b sequence

Characterization

Figure 1: Characterization of CD79b overexpression in the CHOK1-CD79b stable clone using FACS.

Figure 2: Characterization of CD79b overexpression in the CHOK1-CD79b stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (1640 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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% 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

Dornan D, Bennett F, Chen Y, Dennis M, Eaton D, Elkins K, French D, Go MA, Jack A, Junutula JR, Koeppen H, Lau J, McBride J, Rawstron A, Shi X, Yu N, Yu SF, Yue P, Zheng B, Ebens A, Polson AG. Therapeutic potential of an anti-CD79b antibody-drug conjugate, anti-CD79b-vc-MMAE, for the treatment of non-Hodgkin lymphoma. Blood. 2009 Sep 24;114(13):2721-9. doi: 10.1182/blood-2009-02-205500. Epub 2009 Jul 24. PMID: 19633198.
Palanca-Wessels MC, Czuczman M, Salles G, Assouline S, Sehn LH, Flinn I, Patel MR, Sangha R, Hagenbeek A, Advani R, Tilly H, Casasnovas O, Press OW, Yalamanchili S, Kahn R, Dere RC, Lu D, Jones S, Jones C, Chu YW, Morschhauser F. Safety and activity of the anti-CD79B antibody-drug conjugate polatuzumab vedotin in relapsed or refractory B-cell non-Hodgkin lymphoma and chronic lymphocytic leukaemia: a phase 1 study. Lancet Oncol. 2015 Jun;16(6):704-15. doi: 10.1016/S1470-2045(15)70128-2. Epub 2015 Apr 27. PMID: 25925619.