KC-3564

293T-cyno-CLDN1-Cell-Line

26405

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Background of 293T-cyno-CLDN1-Cell-Line

The CLDN1 gene is a crucial "barrier gene." It encodes the Claudin-1 protein, which is essential for forming tight junctions between cells, acting like a "smart wall" that protects our body's internal environment. Whether it functions properly is directly related to skin health, liver function, and the ability to resist pathogens and cancer invasion. In-depth research on it not only helps us understand the pathogenesis of various diseases but also provides important targets for developing new treatments.

Specifications

Catalog Number	KC-3564
Cell Line Name	293T-cyno-CLDN1-Cell-Line
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous cyno-CLDN1 gene.
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-cyno-CLDN1-cell-line was generated using a lentiviral vector expressing the cyno-CLDN1 sequence.

Characterization

Figure 1: Characterization of cyno-CLDN1 overexpression in the 293T-cyno-CLDN1 stable clone using FACS.

Figure 2: Characterization of cyno-CLDN1 in the 293T-cyno-CLDN1 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1. Ghosh U, Agrawal A, Srinivasan VM, Manisha R, Shukla U, Jain V, Nilay M, Kumar H. Neonatal ichthyosis-sclerosing cholangitis syndrome caused by a novel CLDN1 mutation: a case report and literature review. Clin Exp Pediatr. 2025 Oct 2. doi: 10.3345/cep.2025.00906. Epub ahead of print. PMID: 41035246.
2.Boichuk S, Bikinieva F, Dunaev P, Galembikova A, Mikheeva E, Valeeva E, Mani S, Khromova N, Kopnin P, Shigapova L, Deviatiiarov R, Shagimardanova E, Ryzhkin S, Sabirov A. Claudin-1 Contributes to Gastrointestinal Stromal Tumors (GIST) Resistance to Imatinib Mesylate (IM) via Regulation of FGFR-Signaling. Int J Mol Sci. 2025 Aug 22;26(17):8138. doi: 10.3390/ijms26178138. PMID: 40943068; PMCID: PMC12428531.
3. Qu Y, Li Z, Tian L, Zhang X, Wang G, Liu R. Claudin-1, a Biomarker of Epithelial-Mesenchymal Transformation, Enhances Lipid Metabolism in Colorectal Cancer Cells to Promote Tumor Metastasis. J Gene Med. 2025 Sep;27(9):e70034. doi: 10.1002/jgm.70034. PMID: 40890958.

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.