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	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% 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.