KC-3169

293T-Human-CD79a-T2A-CD79b Cell Line

34172

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Background of 293T-Human-CD79a-T2A-CD79b Cell Line

CD79 is a heterodimer molecule that is involved in signal transduction as a component of the B-cell receptor (BCR) and consists of two peptide chains, CD79a (also known as Igα) and CD79b (also known as Igβ). CD79a is one of the signaling components of the pre-B-cell receptor (preBCR). Recent studies have found that CD79a is associated with central nervous system infiltration and relapse in patients with B-cell precursor acute lymphoblastic leukemia (BCP). CD79b is a B cell surface antigen and a component of BCR. CD79b is expressed on the surface of almost all B cells and plays an important role in the expression and transport of BCR to the cell membrane. CD79b is highly expressed on a variety of B cell NHL and chronic lymphocytic leukemia B cells, which has a broad mechanism of killing B cell tumors. CD79a and CD79b also synergistically regulate apoptosis of mature B cells. Therefore, CD79a and CD79b play an important role in the internalization and presentation of BCR. A CD79a-targeted drug, bbT-369, is in phase 2 clinical study. Polatuzumab vedotin, a drug targeting CD79b, has been approved, and a variety of drugs are in clinical research.

Specifications

Catalog Number	KC-3169
Cell Line Name	293T-Human-CD79a-T2A-CD79b Cell Line
Host Cell Line	293T
Description	Stable HEK293T cell line expressing exogenous human CD79a and 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:4~1:5 every 2~3 days; seed out at about 1-3 x 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

293T Human CD79a-T2A-CD79b cell line was generated using a lentiviral vector expressing the human CD79a-T2A CD79b sequence.

Characterization

Figure 1: Characterization of human CD79a and CD79b overexpression in 293T-Human-CD79a-T2A-CD79b stable clones using FACS.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1ug/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 x 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 x 105 cells/ml.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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 x g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3 x106 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

1.Huse K, Bai B, Hilden VI, Bollum LK, Våtsveen TK, Munthe LA, Smeland EB, Irish JM, Wälchli S, Myklebust JH. Mechanism of CD79A and CD79B Support for IgM+ B Cell Fitness through B Cell Receptor Surface Expression. J Immunol. 2022 Nov 15;209(10):2042-2053. doi: 10.4049/jimmunol.2200144. PMID: 36426942; PMCID: PMC9643646. 2.Lenk L, Carlet M, Vogiatzi F, Spory L, Winterberg D, Cousins A, Vossen-Gajcy M, Ibruli O, Vokuhl C, Cario G, El Ayoubi O, Kramer L, Ritgen M, Brüggemann M, Häsler R, Schrappe M, Fuhrmann S, Halsey C, Jeremias I, Hobeika E, Jumaa H, Alsadeq A, Schewe DM. CD79a promotes CNS-infiltration and leukemia engraftment in pediatric Bcell precursor acute lymphoblastic leukemia. Commun Biol. 2021 Jan 15;4(1):73. doi: 10.1038/s42003-020-01591-z. PMID: 33452446; PMCID: PMC7810877.