KC-3471

293T-NFAT-Luc2-GNRHR Cell Line

34412

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Background of 293T-NFAT-Luc2-GNRHR Cell Line

GNRHR (Gonadotropin Releasing Hormone Receptor) is a Protein Coding gene. Diseases associated with GNRHR include Hypogonadotropic Hypogonadism 7 With Or Without Anosmia and Hypogonadotropic Hypogonadism. Among its related pathways are GPCR downstream signalling and Class A/1 (Rhodopsin-like receptors). Gene Ontology (GO) annotations related to this gene include G protein-coupled receptor activity and gonadotropin-releasing hormone receptor activity.

Specifications

Catalog Number	KC-3471
Cell Line Name	293T-NFAT-Luc2-GNRHR Cell Line
Host Cell Line	293T-NFAT-Luc2
Description	Stable 293T-NFAT-Luc2 cell line expressing GNRHR and exogenous luciferase under the control of NFAT signaling pathway.
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 + 150μg/mL Hygromycin B + 1μg/mL Puromycin
Selection Marker	Hygromycin B, Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1 : 10 every 3 days; seed out at about 1 × 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

293T-NFAT-Luc2-GNRHR Cell Line was generated using a lentiviral vector expressing the GNRHR sequence.

Characterization

Figure1: 293T-NFAT-Luc2-GNRHR cells were seeded into 96-well plates, treated with LH-RH for 16 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 150μg/mL Hygromycin B and 1μg/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 : 10 every 3 days; seed out at about 1 × 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

1. Kottler ML, Lorenzo F, Bergametti F, Commerçon P, Souchier C, Counis R. Subregional mapping of the human gonadotropin-releasing hormone receptor (GnRH-R) gene to 4q between the markers D4S392 and D4S409. Hum Genet. 1995 Oct;96(4):477-80. doi: 10.1007/BF00191810. PMID: 7557974.
2. Trarbach EB, Silveira LG, Latronico AC. Genetic insights into human isolated gonadotropin deficiency. Pituitary. 2007;10(4):381-91. doi: 10.1007/s11102-007-0061-7. PMID: 17624596.

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.