KC-5701

U937-TREM1 Cell Line

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Background of U937-TREM1 Cell Line

TREM1 (Triggering Receptor Expressed On Myeloid Cells 1) is a Protein Coding gene, It encodes a receptor belonging to the Ig superfamily that is expressed on myeloid cells. Triggering receptor expressed on myeloid cells 1 (TREM1) is a cell surface receptor expressed on neutrophils, monocytes and some tissue macrophages, where it functions as an immunoregulator that controls myeloid cell responses. The activation of TREM1 is suggested to be an upregulation-based, ligands-induced and structural multimerization-mediated process, in which damage- and pathogen-associated molecular patterns play important roles. Activated TREM1 initiates an array of downstream signaling pathways that ultimately result in the production of pro-inflammatory cytokines and chemokines, whereby it functions as an amplifier of inflammation and is implicated in the pathogenesis of many inflammation-associated diseases. The triggering receptor expressed on myeloid cell 1 (TREM1) plays a critical role in development of chronic inflammatory disorders and the inflamed tumor microenvironment (TME) associated with most solid tumors. Diseases associated with TREM1 include Maxillary Sinusitis and Pneumonia.

Specifications

Catalog Number	KC-5701
Cell Line Name	U937-TREM1 Cell Line
Clone Number	30#
Host Cell Line	Human U937 cell line
Description	Stable U937 cell line expressing exogenous human TREM1 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture 1:2-1:5 every 2-3 days; seed out at about 3-5 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30-60 hours
Mycoplasma Status	Negative

Cell Line Generation

U937-TREM1 cell line was generated using a lentiviral vector expressing the human TREM1 sequence.

Characterization

Figure 1: Figure 1: Characterization of human TREM1 overexpression in the U937-TREM1 stable clone using FACS.

Figure 2: Characterization of TERM1 in the U937 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 + 10% FBS + 1μg/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 1:2-1:5 every 2-3 days; seed out at about 3-5 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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. Li C, Cai C, Xu D, Chen X, Song J. TREM1: Activation, signaling, cancer and therapy. Pharmacol Res. 2024 Jun;204:107212. doi: 10.1016/j.phrs.2024.107212. Epub 2024 May 13. PMID: 38749377.
2. Sun H, Feng J, Tang L. Function of TREM1 and TREM2 in Liver-Related Diseases. Cells. 2020 Dec 7;9(12):2626. doi: 10.3390/cells9122626. PMID: 33297569; PMCID: PMC7762355.
3. Ajith A, Mamouni K, Horuzsko DD, Musa A, Dzutsev AK, Fang JR, Chadli A, Zhu X, Lebedyeva I, Trinchieri G, Horuzsko A. Targeting TREM1 augments antitumor T cell immunity by inhibiting myeloid-derived suppressor cells and restraining anti-PD-1 resistance. J Clin Invest. 2023 Nov 1;133(21):e167951. doi: 10.1172/JCI167951. PMID: 37651197; PMCID: PMC10617775.
4. Li H, Yu W, Zheng X, Zhu Z. TREM1-Microglia crosstalk: Neurocognitive disorders. Brain Res Bull. 2025 Jan;220:111162. doi: 10.1016/j.brainresbull.2024.111162. Epub 2024 Dec 5. PMID: 39645047.