KC-6833

293T-ENPP3 Cell Line

67010

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Background of 293T-ENPP3 Cell Line

ENPP3 encodes ectonucleotide pyrophosphatase/phosphodiesterase 3 and is also known as B10. These ectoenzymes possess ATPase and ATP pyrophosphatase activities and are type II transmembrane proteins. Expression of the related rat mRNA has been found in a subset of immature glial cells and in the alimentary tract. The corresponding rat protein has been detected in the pancreas, small intestine, colon, and liver. The human mRNA is expressed in glioma cells, prostate, and uterus. Expression of the human protein has been detected in uterus, basophils, and mast cells. Two transcript variants, one protein coding and the other non-protein coding, have been found for this gene.

Specifications

Catalog Number	KC-6833
Cell Line Name	293T-ENPP3 Cell Line
NCBI/UniProt Accession Number	NM_005021.5
Clone Number	12#
Host Cell Line	293T
Description	293T-ENPP3 is a stable engineered cell line expressing human ENPP3
Quantity	One vial of frozen cells (≥2×10⁶ cells/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 at a ratio of 1:5–1:8 every 2–3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Estimated 16–31 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T-ENPP3 cell line was generated using a lentiviral vector expressing the human ENPP3 sequence.

Characterization

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:5–1:8 every 2–3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

Tsai SH, Takeda K Regulation of allergic inflammation by the ectoenzyme E-NPP3 (CD203c) on basophils and mast cells. Seminars in immunopathology. 2016 Sep;38(5):571-9. PMID: 27130555. doi: 10.1007/s00281-016-0564-2.
Bühring HJ, Streble A, Valent P The basophil-specific ectoenzyme E-NPP3 (CD203c) as a marker for cell activation and allergy diagnosis. International archives of allergy and immunology. 2004 Apr;133(4):317-29. PMID: 15031605. doi: 10.1159/000077351.
Okada R, Mendoza A, Nousome D et al. ENPP3 CAR T cells combined with CD206 modulation suppress adrenocortical carcinoma. Journal for immunotherapy of cancer. 2026 Jul 1;14(7):. PMID: 42386343. doi: 10.1136/jitc-2025-013726.

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.