KC-3437

CHOK1-rat-GPRC5D-Cell-Line

26391

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Background of CHOK1-rat-GPRC5D-Cell-Line

The orphan G protein–coupled receptor class C group 5 member D (GPRC5D) is a relatively novel target for multiple myeloma immunotherapy. It is an orphan G protein–coupled receptor with unknown function that is highly expressed on malignant bone marrow plasma cells, as well as hard keratinized structures that include the hair shaft, nail, and central region of the tongue. High expression of GPRC5D has been associated with poor prognosis in multiple myeloma. GPRC5D has been used as a target in CAR-T therapy for multiple myeloma, with promising results in preclinical studies. At present, JNJ-64407564 (talquetamab), LM-305, Autologous anti-BCMA/GPRC5D CAR-T cell therapy and other drugs have entered the clinical research stage.

Specifications

Catalog Number	KC-3437
Cell Line Name	CHOK1-rat-GPRC5D-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous rat-GPRC5D gene
Quantity	Two vials 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%RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 every 2-3 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

CHOK1-rat-GPRC5D-cell-line was generated using a lentiviral vector expressing the rat-GPRC5D sequence.

Characterization

Figure 1: Characterization of rat-GPRC5D overexpression in the CHOK1-rat-GPRC5D stable clone using FACS

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70%RPMI 1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1.Lancman G, Sastow DL, Cho HJ, Jagannath S, Madduri D, Parekh SS, Richard S, Richter J, Sanchez L, Chari A. Bispecific Antibodies in Multiple Myeloma: Present and Future. Blood Cancer Discov. 2021 Aug 17;2(5):423-433. doi: 10.1158/2643-3230.BCD-21-0028. PMID: 34661161; PMCID: PMC8510808.