KC-0960

B16/F10-mDDR1-KO-1B3-Cell-Line

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Background of B16/F10-mDDR1-KO-1B3-Cell-Line

Discoidin domain receptor 1 (DDR1) is a collagen-activated receptor tyrosine kinase. Upon collagen binding, DDR1 undergoes tyrosine autophosphorylation, which consequently triggers downstream genetic and cellular pathways and plays critical roles in the regulation of cellular morphogenesis, differentiation, proliferation, adhesion, migration, and invasion. Its expression is mainly limited to epithelial cells located in several organs including skin, kidney, liver and lung. Increasing evidence suggests the potential roles of DDR1 in various human diseases including cancer, fibrosis, atherosclerosis, and other inflammatory disorders. Modulating the activity of DDR1 may be considered as a new therapeutic strategy for human cancer and inflammation-related diseases.

Specifications

Catalog Number	KC-0960
Cell Line Name	B16/F10-mDDR1-KO-1B3-Cell-Line
Host Cell Line	B16/F10
Description	Stable B16/F10 clone with mouse DDR1 gene knockout, No.1B3
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	DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS
Selection Marker	N/A
Morphology	mixture of spindle-shaped and epithelial-like cells
Subculture	Split saturated culture 1:5-1:8 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 17 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

B16/F10-mDDR1-KO-1B3 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of B16/F10-mDDR1-KO-1B3 cell line stable clone using PCR sequencing.

Figure 2: Characterization of B16/F10-mDDR1-KO-1B3 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of B16/F10-mDDR1-KO-1B3 cell line stable clone using western blot.

Cell Resuscitation

Prewarm culture medium (DMEM + 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 1:5-1:8 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

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.