KC-6556

THP-1-CLEC1A-DSRed Cell Line

64154

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Background of THP-1-CLEC1A-DSRed Cell Line

CLEC1A encodes a member of the C-type lectin/C-type lectin-like domain (CTL/CTLD) superfamily. Members of this family share a common protein fold and have diverse functions, such as cell adhesion, cell-cell signaling, glycoprotein turnover, and roles in inflammation and immune response. The encoded protein may play a role in regulating dendritic cell function. This gene is closely linked to other CTL/CTLD superfamily members on chromosome 12p13 in the natural killer gene complex region. Alternative splicing results in multiple transcript variants.

Specifications

Catalog Number	KC-6556
Cell Line Name	THP-1-CLEC1A-DSRed Cell Line
NCBI/UniProt Accession Number	NM_016511.4
Clone Number	7#
Host Cell Line	THP-1
Description	THP-1 cell line stably expressing exogenous CLEC1A-DSRed gene
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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 1 μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Suspension
Subculture	Split saturated culture 1:3-1:6 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 60 hours
Mycoplasma Status	Negative

Cell Line Generation

THP-1-CLEC1A-DSRed cell line was generated using a lentiviral vector expressing the CLEC1A-DSRed sequence.

Characterization

Figure 1: Characterization of CLEC1A overexpression in THP-1-CLEC1A-DSRed stable clones using FACS.

Figure 2: Characterization of DSRed overexpression in THP-1-CLEC1A-DSRed stable clones using FACS.

Figure 3: Characterization of THP-1-CLEC1A-DSRed cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 1 μg/mL puromycin) 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 9.0 mL 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 1:3-1:6 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Zhu G, Lyu L, Yang H, Lee J, Sun J, Zhang J, Xue S, Yan H, Wang L, Chen X, Che C. CLEC-1 Acts as a Negative Regulator of Dectin-1 Induced Host Inflammatory Response Signature in Aspergillus fumigatus Keratitis. Invest Ophthalmol Vis Sci. 2021 May 3;62(6):28. doi: 10.1167/iovs.62.6.28. PMID: 34043748; PMCID: PMC8164365.

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.