KC-6350

293T-rabbit-PTH1R Cell Line

63966

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Background of 293T-rabbit-PTH1R Cell Line

PTH1R (Parathyroid Hormone 1 Receptor) is a Protein Coding gene. After the initial signaling action of parathyroid hormone (PTH) on bone was shown to be activation of adenylyl cyclase, its target was found to be cells of the osteoblast lineage, to the exclusion of osteoclasts and their precursors. This is in addition to the effect of PTH1Rv signaling throughout the osteoblast differentiation process to favour the formation of bone-forming osteoblasts. Initial signaling in the PTH target cells through cAMP and protein kinase A (PKA) activation is extremely rapid, and marked by an amplification process in which the later event, PKA activation, precedes cAMP accumulation in time and is achieved at lower concentrations. PTH-related protein (PTHrP), that was discovered as a cancer product, shares structural similarity with PTH in the amino-terminal domain that allows the hormone, PTH, and the autocrine/paracrine agent, PTHrP, to share actions upon a common G protein coupled receptor, PTH1R, through which they activate adenylyl cyclase with equivalent potencies. Diseases associated with PTH1R include Chondrodysplasia, Blomstrand Type and Metaphyseal Chondrodysplasia, Jansen Type.

Specifications

Catalog Number	KC-6350
Cell Line Name	293T-rabbit-PTH1R Cell Line
NCBI/UniProt Accession Number	NM_001082089.1
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous rabbit PTH1R 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-rabbit-PTH1R cell line was generated using a lentiviral vector expressing the rabbit PTH1R sequence.

Characterization

Figure 1: Characterization of rabbit PTH1R overexpression in the 293T-rabbit-PTH1R stable clone using QPCR.

Figure 2: Characterization of rabbit PTH1R in the 293T-rabbit-PTH1R 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. Obiezu F, Boyce A, Jüppner H, Jha S. PTH1R-Related Jansen Metaphyseal Chondrodysplasia. 2025 Jul 10. In: Adam MP, Bick S, Mirzaa GM, Pagon RA, Wallace SE, Amemiya A, editors. GeneReviews® [Internet]. Seattle (WA): University of Washington, Seattle; 1993–2026. PMID: 40638774..
2. Portales-Castillo I, Höppner J, Jüppner H, Gardella TJ. Human diseases caused by homozygous PTH1R mutations. Front Endocrinol (Lausanne). 2025 Aug 19;16:1641292. doi: 10.3389/fendo.2025.1641292. PMID: 40904804; PMCID: PMC12401709.
3. Yang Y, Wang B. PTH1R-CaSR Cross Talk: New Treatment Options for Breast Cancer Osteolytic Bone Metastases. Int J Endocrinol. 2018 Jul 29;2018:7120979. doi: 10.1155/2018/7120979. PMID: 30151009; PMCID: PMC6087585.

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.