KC-4467

293T-mouse-CD4-Cell-Line

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Background of 293T-mouse-CD4-Cell-Line

The CD4 gene encodes the CD4 membrane glycoprotein of T lymphocytes. CD4 antigen acts as a co-receptor with T cell receptors on T lymphocytes to recognize antigens displayed by antigen-presenting cells in the context of Class II MHC molecules. CD4 antigen is also a major receptor that enters the human immunodeficiency virus through interaction with the HIV Env gp120 subunit. The gene is expressed not only in T lymphocytes, but also in B cells, macrophages, granulocytes, and in various regions of the brain.

Specifications

Catalog Number	KC-4467
Cell Line Name	293T-mouse-CD4-Cell-Line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous mouse CD4 gene
Quantity	One vial 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 mouse CD4 cell line was generated using a lentiviral vector expressing the mouse CD4 sequence.

Characterization

Figure 1: Characterization of mouse CD4 in the 293T mouse CD4 stable clone using PCR sequencing.

Figure 2: Characterization of mouse CD4 overexpression in the 293T mouse CD4 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

Ruterbusch M, Pruner KB, Shehata L, Pepper M. In Vivo CD4⁺ T Cell
Differentiation and Function: Revisiting the Th1/Th2 Paradigm. Annu Rev Immunol.
2020 Apr 26;38:705-725. doi: 10.1146/annurev-immunol-103019-085803. PMID:
Dimeric arrangement of the parathyroid hormone receptor and a structural mechanism for ligand-induced dissociation. PMID: 20172855 PMCID: PMC2852981 DOI: 10.1074/jbc.M109.093138.
Tang H, Jiang X, Zhang J, Pei C, Zhao X, Li L, Kong X. Teleost
CD4⁺ helper T cells: Molecular characteristics and functions and
comparison with mammalian counterparts. Vet Immunol Immunopathol. 2021
Oct;240:110316. doi: 10.1016/j.vetimm.2021.110316. Epub 2021 Aug 25. PMID:
Lange T, Born J, Westermann J. Sleep Matters: CD4⁺ T Cell Memory
Formation and the Central Nervous System. Trends Immunol. 2019
Aug;40(8):674-686. doi: 10.1016/j.it.2019.06.003. Epub 2019 Jun 28. PMID:

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.