KC-3720

293T-cyno-dectin1b Cell Line

34660

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Background of 293T-cyno-dectin1b Cell Line

Dectin-1 is one of the most intensively studied C-type lectin receptors. Dectin-1 is mainly expressed on activated myeloid cells, and as a pattern recognition receptor, dectin-1 is crucial for the regulation of immune homeostasis. It mainly exists in the cell channels that pathogens easily invade, mediates pathogen recognition and phagocytosis, and plays an important role in host defense. The Dectin-1 signaling pathway is involved in the regulation of a variety of cellular responses, including phagocytosis, autophagy, respiratory outbreaks, production of inflammatory lipids, and production of multiple cytokines and chemokines.

Specifications

Catalog Number	KC-3720
Cell Line Name	293T-cyno-dectin1b Cell Line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous cyno dectin1b 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% 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 cyno dectin1b Cell Line was generated using lentivirus expressing cynomolgus dectin1b gene sequence.

Characterization

Figure 1: Characterization of cyno dectin1b overexpression in the 293T cyno dectin1b stable clone using PCR sequence

Figure 2: Characterization of cyno dectin1b overexpression in the 293T cyno dectin1b stable clone using qPCR.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1ug/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 x 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 1:4 ~ 1:5 every 2~3 days; seed out at about 1-3 x 10 5 cells/ml.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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 x g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3 x106 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. Moerings BGJ, van Bergenhenegouwen J, Furber M, Abbring S, Schols HA, Witkamp RF, Govers C, Mes JJ. Dectin-1b activation by arabinoxylans induces trained immunity in human monocyte-derived macrophages. Int J Biol Macromol. 2022 Jun 1;209(Pt A):942-950. doi: 10.1016/j.ijbiomac.2022.04.071. Epub 2022 Apr 18. PMID: 35447262. 2. Batista PJ, Nuzzo G, Gallo C, Carbone D, dell'Isola M, Affuso M, Barra G, Albiani F, Crocetta F, Virgili R, Mazzella V, Castiglia D, d'Ippolito G, Manzo E, Fontana A. Chemical and Pharmacological Prospection of the Ascidian Cystodytes dellechiajei. Mar Drugs. 2024 Jan 31;22(2):75. doi: 10.3390/md22020075. PMID: 38393046; PMCID: PMC10890457. 3. Grünebach F, Weck MM, Reichert J, Brossart P. Molecular and functional characterization of human Dectin-1. Exp Hematol. 2002 Nov;30(11):1309-15. doi: 10.1016/s0301-472x(02)00928-1. PMID: 12423684.