C12 Cell Line Overexpressing Human P2X3

Tool Details

*FOR RESEARCH USE ONLY (for other uses, please contact the licensing team)

• Name: C12 Cell Line Overexpressing Human P2X3
• Research fields: Cell biology
• Parental cell: HEK293
• Organism: Human
• Gender: Female
• Tissue: Kidney
• Morphology: Epithelial morphology, typically less circular than other common cell lines, and they often display dendritic-like processes. They are slightly granular, and they tend to be adherent.
• Growth properties: Adherent
• Model: Genetically modified cell line
• Crispr: No
• Description: Cell Line is a clone that expresses hP2X3 in a HEK293 cell background. Currents evoked by ATP are reliable and of high amplitude.
• Application: Patch-clamp recording and calcium uptake assay
• Production details: When the cell culture reaches 90% confluence, the cells need to be passaged. So, cell culture medium is removed, wash once with PBS, and then treated with enough but minimun amount of 0.05% trypsin for 2 min, inside a 37C and 5% CO2 incubator. Trypsin activity is stop by addition of 10 times volume of complete cell culture medium. Aliquots of cell culture (1x10^6 cells in 1 ml frozen media per vial) can be frozen back for long time storage in liquid nitrogen. Number of cell passages can be minized by subculturing the cells once a week, with the right amount of cells according to the size of the culture dish or flask used.
• Recommended controls: Wildtype HEK293 cells

Applications

• Application: Patch-clamp recording and calcium uptake assay

Handling

• Format: Frozen
• Passage number: Passage 44 (P44)
• Growth medium: DMEM medium with 10% FBS, 1% Penicillin/Streptomycin, and G-418 (0.6 mg/ml).
• Temperature: 37° C
• Atmosphere: 5% CO2
• Shipping conditions: Dry ice
• Storage medium: Human P2X3 receptor channel
• Initial handling information: The vial containing the frozen cells is warm up in a bath. Cells are resuspended before the storage medium containing the cells are completely thawed, with 1 ml of culture cell media. The resuspension is transferred to a 15 ml tube and 8 ml of cell culture media is added and gently resuspended again. The cell suspension is centriguged at 1000 rpm for 5 min and the superntant discarded. The cells pellet is resuspended in 2 ml of cell culture media and seed it in a T-75 flask with 20 ml final volume of media. The day after, cell culture media is removed to eliminate death cells and replaced by a fresh cell culture media. If the culture is confluent, then it needs to be passaged. So, cells will be treated with 0.05% trypsin for 2 min inside a 37C and 5% CO2 incubator to dettach the cells from the botom of the flask. A gentle blow of the flask will warranty all the cells to be dettached. Check it out under the microscope.
• Cultured in antibiotics: Penicillin, Streptomycin, G-418.
• Characterisation tests: Patch-clamp recording; Ratiometric calcium uptake assay.