Cat.No. : CSC-RI0057 Host Cell: HEK293
| Cat. No. | CSC-RI0057 |
| Description | This cell line is engineered to overexpress human TRPV1. |
| Gene | TRPV1 |
| Gene Species | Homo sapiens (Human) |
| Alias | TRPV1, OTRPC1, VR1, DKFZp434K0220 |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Morphology | Epithelial |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Channelopathies research |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:3-1:6. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
Transient receptor potential vanilloid 1 (TRPV1) is a noxious, thermosensitive, chemo-nociceptive cation channel expressed in primary sensory neurons of multimodal nociceptors. The present study was devoted to analyzing the role of lipid raft components in calcium influx induced by various TRPV1 agonists in sensory neurons and rTRPV1-transfected CHO cell lines. Depletion of cholesterol by methyl β-cyclodextrin (MCD, 1-10 mM) reduced the percentage of calcium uptake in cultured trigeminal neurons in response to capsaicin or resiniferatoxin (RTX, 3 nM). In contrast, in TRPV1-transfected cells, inhibition was observed only when capsaicin or N-oleoyl dopamine (OLDA, 10 µM) was applied, but not when gated with RTX, anandamide (AEA, 10 µM), or pH 5.5. Treatment of rTRPV1-expressing cells with sphingomyelinase inhibited capsaicin-induced 45Ca uptake, whereas RTX-induced responses remained unchanged. On the other hand, in trigeminal neurons, the effects of both compounds were inhibited by sphingomyelinase treatment. Inhibition of ganglioside biosynthesis by D-threo-1-phenyl-2-decanoylamino-3-morpholine-1-propanol (D-PDMP) or myriocyn also reduced calcium uptake induced by capsaicin or RTX in cultured trigeminal neurons and rTRPV1-expressing cells. The present study demonstrates that depletion of different components of lipid rafts inhibits gating of TRPV1 cation channels by various vanilloid and non-vanilloid agents. Evidence for supporting roles of cholesterol, sphingomyelin and gangliosides was obtained in both native and TRPV1-transfected cells.
Figure 1. Effect of combined administration of low concentrations of MCD and SMase on capsaicin- and RTX-induced Ca2+-accumulation. A: Results of radioactive 45Ca-uptake experiments in TRPV1-expressing CHO cell line. Ca2+-influx is presented in % of control (untreated). B: Results of the same pretreatment on cultured trigeminal sensory neurons. Percentage of responding cells to capsaicin and RTX is presented (responsive cells vs. non-responsive cells). (Szőke É, et al. 2010)
Creative Biogene has successfully constructed TRPV1 over-expressing stable cell lines with HEK293, CHO, HeLa, 1321N1, U2OS, RH7777, etc. These TRPV1 stable cell lines have excellent in vitro assay sensitivity and reproducibility. Therefore, conducting experiments with our TRPV1 stable cell lines saves time and resources while providing more accurate results.
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I have been using the Human TRPV1 Stable Cell Line-HEK293 for my pain and sensation research projects with excellent results.
As a researcher studying pain perception, the Human TRPV1 Stable Cell Line-HEK293 is extremely valuable. The stable expression of TRPV1 in these cells ensures consistent and reliable results for electrophysiological assays and calcium imaging studies.
Using the Human TRPV1 Stable Cell Line-HEK293 has considerably improved my project on thermal pain sensation. The cell culture is easy to maintain, and the expression of TRPV1 remains consistent over multiple passages.
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