KC-6202

E0771-mDDR1-KO Cell Line

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Background of E0771-mDDR1-KO Cell Line

The mouse discoidin domain receptor 1 (mDDR1) gene, localized on chromosome 1q23.3, encodes a transmembrane receptor tyrosine kinase (RTK) belonging to the discoidin domain receptor family. This receptor is characterized by an extracellular discoidin domain that mediates binding to collagens, its primary ligands. mDDR1 plays pivotal roles in regulating cell adhesion, migration, proliferation, and matrix remodeling, with critical implications in embryonic development, tissue repair, and pathological processes such as fibrosis and cancer progression. Functionally, mDDR1 signaling intersects with key pathways including the MAPK/ERK and PI3K/Akt cascades, modulating cellular responses to extracellular matrix cues. Studies using mDDR1 knockout mice have elucidated its non-redundant functions in tissue homeostasis—particularly in the kidney, mammary gland, vascular system, and nervous system—and its roles in regulating collagen ultrastructure, platelet adhesion, and neuronal axon extension.

Specifications

Catalog Number	KC-6202
Cell Line Name	E0771-mDDR1-KO Cell Line
NCBI/UniProt Accession Number	12305
Clone Number	1B1
Host Cell Line	E0771
Description	Stable E0771-mDDR1-KO cell line with mDDR1 gene knockout, No.1B1
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
Selection Marker	NA
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

E0771-mDDR1-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of E0771-mDDR1-KO Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of E0771-mDDR1-KO Cell Line stable clone using RT-PCR sequencing.

Figure 3: Characterization of E0771-mDDR1-KO 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 1:4-1:8 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

Chavez MB, Kolli TN, Tan MH, Zachariadou C, Wang C, Embree MC, Lira Dos Santos EJ, Nociti FH Jr, Wang Y, Tatakis DN, Agarwal G, Foster BL. Loss of Discoidin Domain Receptor 1 Predisposes Mice to Periodontal Breakdown. J Dent Res. 2019 Dec;98(13):1521-1531. doi: 10.1177/0022034519881136. Epub 2019 Oct 14. PMID: 31610730; PMCID: PMC6873285.
Bhatt RS, Tomoda T, Fang Y, Hatten ME. Discoidin domain receptor 1 functions in axon extension of cerebellar granule neurons. Genes Dev. 2000 Sep 1;14(17):2216-28. doi: 10.1101/gad.821600. PMID: 10970885; PMCID: PMC316891.