KC-3086

293T-CD79a Cell Line

34112

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Background of 293T-CD79a Cell Line

After the antigen binds to the BCR, the CD79A/CD79B heterodimer can transmit activation signals, triggering the proliferation, differentiation and antibody production of B cells. The cytoplasmic region of CD79A contains the immunoreceptor tyrosine-based activation motif (ITAM), which is a key structure for signal transduction. The important regulatory role of CD79A in B cell function is involved in the processes of antigen recognition, activation, proliferation and differentiation of B cells. The expression of CD79A is also associated with the occurrence and development of certain B cell tumors, such as B-cell acute lymphoblastic leukemia (B-ALL).

Specifications

Catalog Number	KC-3086
Cell Line Name	293T-CD79a Cell Line
Host Cell Line	293T
Description	Stable HEK293T cell line expressing exogenous human CD79a 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 + 1ug/ml Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid cells growing as a monolayer
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	NA

Cell Line Generation

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

Characterization

Figure 1: Characterization of CD79a overexpression in 293T-CD79a stable clones using FACS.

Figure 2: Characterization of endogenous human CD79a expression in 293T using FACS.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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 × 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 1:4-1:8 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.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. 2.Hashimoto M, Yamashita Y, Mori N. Immunohistochemical detection of CD79a expression in precursor T cell lymphoblastic lymphoma/leukaemias. J Pathol. 2002 Jul;197(3):341-7. 3.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.