KC-6034

JJN-3-Dectin-1-KO cell line

60545

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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-6034
Cell Line Name	JJN-3-Dectin-1-KO cell line
Clone Number	1C6
Host Cell Line	JJN-3
Description	Stable JJN-3-Dectin-1-KO cell clone with human Dectin-1 gene knockout
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/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.

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.