KC-4019

293T-CDCP1-High-Cell-Line

40496

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Background of 293T-CDCP1-High-Cell-Line

CDCP1, also known as CD318, encodes a transmembrane protein that contains three extracellular CUB domains and serves as a substrate for Src family kinases. This protein plays a role in tyrosine phosphorylation dependent regulation of cellular events involved in tumor invasion and metastasis.

Specifications

Catalog Number	KC-4019
Cell Line Name	293T-CDCP1-High-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous CDCP1 gene in high level
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
In Vivo Validation	NA

Cell Line Generation

293T-CDCP1-High-cell-line was generated using a lentiviral vector expressing the CDCP1 sequence.

Characterization

Figure 1: Characterization of CDCP1 overexpression in the 293T-CDCP1-High stable clone using FACS.

Figure 2: Characterization of CDCP1 in the 293T-CDCP1 stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)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

Qi X, Gao J, Li Z, Zhang G, Li J, Fu Y, Cai M, Wang H, Tong T. CDCP1: A promising diagnostic biomarker and therapeutic target for human cancer. Life Sci. 2022 Jul 15;301:120600. doi: 10.1016/j.lfs.2022.120600. Epub 2022 May 2. PMID: 35504333.
Lim SA, Zhou J, Martinko AJ, Wang YH, Filippova EV, Steri V, Wang D, Remesh SG, Liu J, Hann B, Kossiakoff AA, Evans MJ, Leung KK, Wells JA. Targeting a proteolytic neoepitope on CUB domain containing protein 1 (CDCP1) for RAS-driven cancers. J Clin Invest. 2022 Feb 15;132(4):e154604. doi: 10.1172/JCI154604. PMID: 35166238; PMCID: PMC8843743.