KC-1985

293T-TREM1-DAP12-Cell-Line

23179

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Background of 293T-TREM1-DAP12-Cell-Line

TREM1, full named as Triggering receptor expressed on myeloid cells 1, is an immunoglobulin (Ig) superfamily transmembrane protein which is expressed on myeloid cells. TREM1 forms a complex with transmembrane adaptor DAP12 and then mediates signaling and cell activation after binding serval ligands. TREM1 expression is higher in tumor vs non-tumor tissues, likely due to its expression by myeloid cells that infiltrate tumors, and is also believed to be a promising target for cancer therapy.

Specifications

Catalog Number	KC-1985
Cell Line Name	293T-TREM1-DAP12-Cell-Line
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous TREM1-DAP12 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
Subculture	Split saturated culture 1:4-1:8 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

293T TERM1-DAP12 cell line was generated using a lentiviral vector expressing the TERM1-DAP12 sequence.

Characterization

Figure 1: Characterization of TREM1 overexpression in 293T stable clones using FACS.

Cell Resuscitation

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

Cell Freezing

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

References

Gingras MC, Lapillonne H, Margolin JF (March 2002). TREM-1, MDL-1, and DAP12 expression is associated with a mature stage of myeloid development. Molecular Immunology. 38 (11): 817–24. doi:10.1016/S0161-5890(02)00004-4. PMID 11922939.
Bouchon A, Dietrich J, Colonna M (May 2000). Cutting edge: inflammatory responses can be triggered by TREM-1, a novel receptor expressed on neutrophils and monocytes. Journal of Immunology. 164 (10): 4991–5. doi:10.4049/jimmunol.164.10.4991.
Broz, Miranda L.; Binnewies, Mikhail; Boldajipour, Bijan; Nelson, Amanda E.; Pollack, Joshua L.; Erle, David J.; Barczak, Andrea; Rosenblum, Michael D.; Daud, Adil; Barber, Diane L.; Amigorena, Sebastian (November 2014). Dissecting the Tumor Myeloid Compartment Reveals Rare Activating Antigen-Presenting Cells Critical for T Cell Immunity. Cancer Cell. 26 (5): 638–652.
Lavin, Yonit; Kobayashi, Soma; Leader, Andrew; Amir, El-ad David; Elefant, Naama; Bigenwald, Camille; Remark, Romain; Sweeney, Robert; Becker, Christian D.; Levine, Jacob H.; Meinhof, Klaus (May 2017). Innate Immune Landscape in Early Lung Adenocarcinoma by Paired Single-Cell Analyses. Cell. 169 (4): 750–765.e17.

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.