KC-5067

293T-mouse-CD127-Cell-Line

57527

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Background of 293T-mouse-CD127-Cell-Line

CD127 (also known as IL-7R alpha) is the alpha chain of the interleukin-7 receptor (IL-7R), and the protein encoded by this gene is the receptor for interleukin-7 (IL7). The function of this receptor requires the interleukin-2 receptor gamma chain (IL2RG), which is a common gamma chain shared by various cytokine receptors, including interleukin-2, 4, 7, 9, and 15. It is an important member of the cytokine receptor family and plays a crucial role in the development and function of the immune system. CD127 is a core target of immune regulation, with multiple roles in development, homeostasis, and disease. Its agonists or antagonists have broad application prospects in the fields of infection, tumors, and autoimmune diseases.

Specifications

Catalog Number	KC-5067
Cell Line Name	293T-mouse-CD127-Cell-Line
Clone Number	1-4#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous mouse-CD127 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-mouse-CD127-cell-line was generated using a lentiviral vector expressing the mouse-CD127 sequence.

Characterization

Figure 1: Characterization of mouse-CD127 overexpression in the 293T-mouse-CD127 stable clone using FACS.

Figure 2: Characterization of mouse-CD127 in the 293T-mouse-CD127 stable clone using PCR sequencing.

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.Lundtoft C, Seyfarth J, Jacobsen M. IL7RA genetic variants differentially affect IL-7Rα expression and alternative splicing: a role in autoimmune and infectious diseases? Genes Immun. 2020 Feb;21(2):83-90. doi: 10.1038/s41435-019-0091-y. Epub 2020 Jan 13. PMID: 31929513.
2.Campos LW, Pissinato LG, Yunes JA. Deleterious and Oncogenic Mutations in the IL7RA. Cancers (Basel). 2019 Dec 5;11(12):1952. doi: 10.3390/cancers11121952. PMID: 31817502; PMCID: PMC6966522.