KC-2177

HEK293T-cAMP-biosensor-22F-H-1B2-Cell-Line

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Background of HEK293T-cAMP-biosensor-22F-H-1B2-Cell-Line

Cyclic adenosine monophosphate (cAMP) is a second messenger that mediates numerous biological responses. Intracellular cAMP levels are increased by activation of G(s)-coupled G protein-coupled receptors (GPCRs) and decreased by activation of G(i)-coupled GPCRs via the adenylyl cyclase. Studies on the cAMP dynamics provided useful scientific insights for drug development and treatment of cAMP-related diseases such as some cancers and prefrontal cortex disorders.

Specifications

Catalog Number	KC-2177
Cell Line Name	HEK293T-cAMP-biosensor-22F-H-1B2-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	HEK293 cell line stable expressing exogenous GloSensor™ luciferase.
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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+100μg/mL Hygromycin B
Selection Marker	HygromycinB
Morphology	Epithelial
Subculture	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
In Vivo Validation	NA

Cell Line Generation

HEK293-cAMP-biosensor-22F luciferase cell line was generated using a lentiviral vector expressing exogenous GloSensor sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 100μg/mL Hygromycin) 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 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Jesper Mosolff Mathiesen , Line Vedel, Hans Br?uner-Osborne. cAMP biosensors applied in molecular pharmacological studies of G protein-coupled receptors. Methods Enzymol. 2013;522:191-207.
Namdoo Kim, Seunghan Shin, and Se Won Bae. cAMP Biosensors Based on Genetically Encoded Fluorescent/Luminescent Proteins. Cardiovascular Toxicology. Biosensors (Basel). 2021 Feb; 11(2): 39.

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.