KC-6035

JJN-3-Dectin-1-KO cell line

60549

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Background of JJN-3-Dectin-1-KO cell line

Dectin-1, also known as β-glucan receptor, is a type II transmembrane lectin belonging to the family of CLRs. Structurally, Dectin-1 has a single CRD that specifically recognizes polysaccharides defined as β- (1 → 3)/(1 → 6) – glucans, initially described to be present in the cell wall of certain pathogens including fungi and some bacteria.

Specifications

Catalog Number	KC-6035
Cell Line Name	JJN-3-Dectin-1-KO cell line
Clone Number	2B5
Host Cell Line	JJN-3
Description	Stable JJN-3-Dectin-1-KO cell clone with human Dectin-1 gene knockout
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/IMDM(1:1) + 20% FBS
Selection Marker	NA
Morphology	Lymphoblast
Subculture	Split the saturated culture at a ratio of 1:3-1:4 every 3-4 days; seed out at about 1-3 x 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

JJN-3-Dectin-1-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of JJN-3-Dectin-1-KO cell line stable clone using PCR sequencing.

Figure 2:Characterization of JJN-3-Dectin-1-KO cell line stable clone using RT-PCR sequencing.

Figure 3:Characterization of JJN-3-Dectin-1-KO cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM/IMDM(1:1) + 20% FBS)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 1:3-1:4 every 3-4 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

Mata-Martínez P, Bergón-Gutiérrez M, Del Fresno C. Dectin-1 Signaling Update: New Perspectives for Trained Immunity. Front Immunol. 2022 Feb 14;13:812148. doi: 10.3389/fimmu.2022.812148. PMID: 35237264; PMCID: PMC8882614.