KC-6459

293T-mIgE-short Cell Line

62640

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Background of 293T-mIgE-short Cell Line

The IGHE(IGE) gene encodes the constant region of the heavy chain of IgE antibodies and is a core component of the IgE molecule. Upregulation of IGHE gene expression is closely related to IgE-mediated allergic diseases, such as allergic rhinitis, asthma, and atopic dermatitis. Monoclonal antibody drugs targeting IgE (such as omalizumab) are used to treat moderate to severe asthma and chronic spontaneous urticaria by blocking the binding of IgE to its receptors.

Specifications

Catalog Number	KC-6459
Cell Line Name	293T-mIgE-short Cell Line
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous mIgE-short 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	Epithelial
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

Cell Line Generation

293T-mIgE-short cell line was generated using lentivirus expressing mIgE-short sequence.

Characterization

Figure 1. Characterization of mIgE-short over-expression in the 293T-mIgE-short stable clone using FACS.

Figure 2. Characterization of 293T-mIgE-short cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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% 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×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

Luo L, Luo Y, Xu J, Zhu R, Wu J, Liu X, Li W, Yao X. Heterogeneous origin of IgE in atopic dermatitis and psoriasis revealed by B cell receptor repertoire analysis. Allergy. 2022 Feb;77(2):559-568. doi: 10.1111/all.15173. Epub 2021 Nov 12. PMID: 34738638.