KC-5659

CHOK1-TMIGD2-Cell-Line

59997

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Background of CHOK1-TMIGD2-Cell-Line

TMIGD2 is an important immune co-stimulatory molecule that participates in regulating the activity of T lymphocytes by interacting with the receptor IGPR-1/CD28H, playing a core role in initiating and maintaining adaptive immune responses. TMIGD2 is mainly expressed on the surface of antigen-presenting cells such as dendritic cells, macrophages, and B cells. Additionally, it is highly expressed on certain types of tumor cells. It has a dual role in tumor immunity, being both a potential immune-enhancing target and possibly an accomplice in immune suppression, making it a "hotspot" for drug development. Antibody drugs targeting the TMIGD2-CD28H pathway as agonists and antagonists are currently in preclinical and early clinical research stages and are strong candidates for the next generation of immune checkpoint inhibitors/agonists, promising to open up new avenues for the treatment of cancer and autoimmune diseases.

Specifications

Catalog Number	KC-5659
Cell Line Name	CHOK1-TMIGD2-Cell-Line
NCBI/UniProt Accession Number	NM_144615
Clone Number	4#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous TMIGD2 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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-TMIGD2-cell-line was generated using a lentiviral vector expressing the TMIGD2 sequence.

Characterization

Figure 1: Characterization of TMIGD2 overexpression in the CHOK1-TMIGD2 stable clone using FACS.

Figure 2: Characterization of TMIGD2 in the CHOK1-TMIGD2 stable clone using PCR sequencing.

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.Rahimi N, Potluri S, Chitalia V. IGPR-1 Is Phosphorylated on the Immunoreceptor Tyrosine-Based Motif, Stimulates the AKT Pathway and Supports Melanoma Growth. Biochemistry. 2025 Oct 7;64(19):4121-4131. doi: 10.1021/acs.biochem.5c00393. Epub 2025 Sep 16. PMID: 40957723.
2.Zhao Y, Chen H, Zhang W, Shang W, Cao J, Zhao H, Zou Z. Mir-615-5p inhibits cervical cancer progression by targeting TMIGD2. Hereditas. 2025 Jan 9;162(1):4. doi: 10.1186/s41065-024-00363-7. PMID: 39789663; PMCID: PMC11715597.
3.Wang J, Wu Y. MicroRNA-486-3p affects cisplatin resistance in high-grade serous ovarian cancer by regulating TMIGD2: An experimental study. Heliyon. 2024 Jul 19;10(15):e34978. doi: 10.1016/j.heliyon.2024.e34978. PMID: 39145009; PMCID: PMC11320304.