KC-4514

CHOK1-CLDN17-Cell-Line

47008

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Background of CHOK1-CLDN17-Cell-Line

CLDN17 (Claudin 17) is a Protein Coding gene, This gene encodes a member of the claudin family. The claudin (CLDN) family, as key components of tight junctions (TJs), plays an important role in the initiation and development of cancer. CLDN17, a putative anion pore-forming CLDN based on its structural characterization, is assumed to regulate anion balance across the blood-tissue barriers. CLDN17 can reduce cell invasion and migration by inhibiting the EMT process, becoming a potential therapeutic target for oral cancer. Diseases associated with CLDN17 include Amyotrophic Lateral Sclerosis 1.

Specifications

Catalog Number	KC-4514
Cell Line Name	CHOK1-CLDN17-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human CLDN17 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-CLDN17 Cell Line was generated using a lentiviral vector expressing the human CLDN17 sequence

Characterization

Figure 1: Characterization of human overexpression in the CHOK1 CLDN17 stable clone using FACS.

Figure 2: Characterization of human CLDN17 in the CHOK1 CLDN17 stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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% RPMI1640 + 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

Adil MS, Parvathagiri V, Verma A, Liu F, Rudraraju M, Narayanan SP, Somanath PR. Claudin-17 Deficiency in Mice Results in Kidney Injury Due to Electrolyte Imbalance and Oxidative Stress. Cells. 2022 May 29;11(11):1782. doi: 10.3390/cells11111782. PMID: 35681477; PMCID: PMC9180152.
Wang W, Zhou Y, Li W, Quan C, Li Y. Claudins and hepatocellular carcinoma. Biomed Pharmacother. 2024 Feb;171:116109. doi: 10.1016/j.biopha.2023.116109. Epub 2024 Jan 6. PMID: 38185042.
Xu YN, Deng MS, Liu YF, Yao J, Xiao ZY. Tight junction protein CLDN17 serves as a tumor suppressor to reduce the invasion and migration of oral cancer cells by inhibiting epithelial-mesenchymal transition. Arch Oral Biol. 2022 Jan;133:105301. doi: 10.1016/j.archoralbio.2021.105301. Epub 2021 Oct 30. PMID: 34781072.

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.