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	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% 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.

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.