KC-3119

CT26-mDDR1-KO-2C2 Cell Line

27151

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Background of CT26-mDDR1-KO-2C2 Cell Line

Discoidin Domain Receptor 1 (DDR1) is a protein that belongs to the discoidin domain receptor family. It is a transmembrane receptor tyrosine kinase that plays an important role in cell adhesion, migration, and proliferation. DDR1 is expressed in various tissues, including the heart, kidney, lung, liver, and brain, and has been implicated in several physiological and pathological processes. For example, DDR1 is involved in the regulation of collagen synthesis and deposition, which is important for tissue remodeling during wound healing and fibrosis. It is also involved in the regulation of cell survival and apoptosis, as well as in the development and progression of cancer. Mutations or dysregulation of DDR1 have been associated with several diseases, including cancer, fibrosis, and cardiovascular disease. As a result, DDR1 has become a potential therapeutic target for the treatment of these diseases.

Specifications

Catalog Number	KC-3119
Cell Line Name	CT26-mDDR1-KO-2C2 Cell Line
Host Cell Line	CT26
Description	Stable CT26 clone with mDDR1 gene knockout, No.2C2
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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640+10%FBS
Selection Marker	NA
Morphology	Fibroblast
Subculture	Split saturated culture 1:4-1:5 every 1-2 days; seed out at about 1 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CT26-mDDR1-KO cell line was generated using CRISPR method.

Characterization

Figure 1: Characterization of CT26-mDDR1-KO-2C2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of CT26-mDDR1-KO-2C2 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of CT26-mDDR1-KO-2C2 Cell Line stable clone using western blot.

Figure 4: Morphology of CT26-mDDR1-KO-2C2 Cell Line.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS) 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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Guo J, Zhang Z, Ding K. A patent review of discoidin domain receptor 1 (DDR1) modulators (2014-present). Expert Opin Ther Pat. 2020 May;30(5):341-350. doi: 10.1080/13543776.2020.1732925. Epub 2020 Feb 26. PMID: 32077340.
Moll S, Desmoulière A, Moeller MJ, Pache JC, Badi L, Arcadu F, Richter H, Satz A, Uhles S, Cavalli A, Drawnel F, Scapozza L, Prunotto M. DDR1 role in fibrosis and its pharmacological targeting. Biochim Biophys Acta Mol Cell Res. 2019 Nov;1866(11):118474. doi: 10.1016/j.bbamcr.2019.04.004. Epub 2019 Apr 5. PMID: 30954571.