KC-4117

293T-MC1R Cell Line

63921

Home » 293T-MC1R Cell Line

Background of 293T-MC1R Cell Line

MC1R (Melanocortin 1 Receptor) is a Protein Coding gene. Melanocortin 1 receptor (MC1R), a G protein-coupled receptor primarily known for regulating skin pigmentation and exhibiting anti-inflammatory effects. Melanocortins play crucial roles in regulating the stress response, inflammation, and skin pigmentation. Activation of MC1R in melanocytes by α-melanocyte-stimulating hormone (α-MSH) stimulates cAMP signalling and melanin production and enhances DNA repair after ultraviolet irradiation. Binding of MSH to its receptor activates the receptor and stimulates eumelanin synthesis. This receptor is a major determining factor in sun sensitivity and is a genetic risk factor for melanoma and non-melanoma skin cancer. Diseases associated with MC1R include Skin/Hair/Eye Pigmentation, Variation In, 2 and Melanoma, Cutaneous Malignant 5.

Specifications

Catalog Number	KC-4117
Cell Line Name	293T-MC1R Cell Line
NCBI/UniProt Accession Number	NM_002386.3
Clone Number	3#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous human MC1R gene
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 +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:5 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-MC1R cell line was generated using a lentiviral vector expressing the human MC1R sequence.

Characterization

Figure 1: Characterization of human MC1R overexpression in the 293T-MC1R stable clone using QPCR.

Figure 2: Characterization of human MC1R in the 293T-MC1R stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (DMEM+10% FBS +1μg/mL Puromycin) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:5 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Guida S, Guida G, Goding CR. MC1R Functions, Expression, and Implications for Targeted Therapy. J Invest Dermatol. 2022 Feb;142(2):293-302.e1. doi: 10.1016/j.jid.2021.06.018. Epub 2021 Aug 4. PMID: 34362555.
2. Herraiz C, Garcia-Borron JC, Jiménez-Cervantes C, Olivares C. MC1R signaling. Intracellular partners and pathophysiological implications. Biochim Biophys Acta Mol Basis Dis. 2017 Oct;1863(10 Pt A):2448-2461. doi: 10.1016/j.bbadis.2017.02.027. Epub 2017 Mar 1. PMID: 28259754..
3. Mun Y, Kim W, Shin D. Melanocortin 1 Receptor (MC1R): Pharmacological and Therapeutic Aspects. Int J Mol Sci. 2023 Jul 29;24(15):12152. doi: 10.3390/ijms241512152. PMID: 37569558; PMCID: PMC10418475.

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.