KC-2645

293T-DDR1 Cell Line

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Background of 293T-DDR1 Cell Line

Discoidin domain receptor 1 (DDR1), a receptor tyrosine kinase activated by collagen, Its expression is mainly limited to epithelial cells located in several organs including skin, kidney, liver and lung. DDR1 plays a crucial role in maintaining intracellular environment homeostasis through metabolic reprogramming. In addition, it has been shown to be significantly overexpressed in several human tumors.

Specifications

Catalog Number	KC-2645
Cell Line Name	293T-DDR1 Cell Line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous DDR1 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 1:4-1:5 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 DDR1 cell line was generated using a lentiviral vector expressing the DDR1 sequence.

Characterization

Figure 1: Characterization of endogenous DDR1 expression in 293T cells using FACS.

Figure 2: Characterization of DDR1 overexpression in the 293T DDR1 stable clone 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 1:4-1:5 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

1. Xiong B, Xie Z, Song F, Chen H, Wang X, Jin Z, Han T, Li Y, Zhang D. DDR1 promotes LoVo cell proliferation by regulating energy metabolism. Acta Biochim Biophys Sin (Shanghai). 2022 May 25;54(5):615-624.
2. Duan X, Xu X, Zhang Y, Gao Y, Zhou J, Li J. DDR1 functions as an immune negative factor in colorectal cancer by regulating tumor-infiltrating T cells through IL-18. Cancer Sci. 2022 Nov;113(11):3672-3685.