CLDN18.2 is the isoform 2 of the claudin 18 gene, which belong to the lager claudin family and play the important role in cell tight junction in epithelial cells. CLND18.2 is found overexpressed on gastrointestinal adenocarcinoma and pancreatic tumors. The identification as a tumor target of CLDN18.2 has led to the repaid progress of antibody treatment of gastrointestinal adenocarcinoma and pancreatic tumors, such as IMAB362 (Claudiximab).
Specifications
Catalog Number:
KC-1651
Cell Line Name:
NUGC4-CLDN18.2-Luc2 Cell Line
Host Cell Line:
Human NUGC4 cell line
Description:
NUGC4 cell line stable expressing exogenous human CLDN18 isoform 2 and luciferase gene
Split saturated culture 1:3-1:6 every 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
Cell Line Generation
NUGC4 CLDN18.2-Luc3 cell line was generated using lentiviral vector expressing human CLDN18.2 sequence.
Characterization
Application
Cell Resuscitation
1. Prewarm culture medium (RPMI1640 + 10% FBS + 75µg/mL Hygromycin B + 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 1:3-1:6 every 3 days; seed out at about 1-3 × 105 cells/mL.
Cell Freezing
1. Prepare the freezing medium (70% RPMI-1640 + 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. Micke, Patrick, Johanna Sofia Margareta Mattsson, Karolina Edlund, Miriam Lohr, Karin Jirstr?m, Anders Berglund, Johan Botling, et al. 2014. ¡ùAberrantly Activated Claudin 6 and 18.2 as Potential Therapy Targets in Non-Small-Cell Lung Cancer.¡ì International Journal of Cancer 135 (9): 2206ÿ14. doi:10.1002/ijc.28857.
2. Singh, Prabhsimranjot, Sudhamshi Toom, and Yiwu Huang. 2017. ¡ùAnti-Claudin 18.2 Antibody as New Targeted Therapy for Advanced Gastric Cancer,¡ì May. Journal of Hematology & Oncology, 1ÿ5. doi:10.1186/s13045-017-0473-4.
3. Sahin, U, M Koslowski, K Dhaene, D Usener, G Brandenburg, G Seitz, C Huber, and O Tureci. 2008. ¡ùClaudin-18 Splice Variant 2 Is a Pan-Cancer Target Suitable for Therapeutic Antibody Development.¡ì Clinical Cancer Research 14 (23): 7624ÿ34. doi:10.1158/1078-0432.CCR-08-1547.
