KC-4019

293T-CDCP1-High-Cell-Line

40496

Home » 293T-CDCP1-High-Cell-Line

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	One vial 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.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.