KC-6559

THP-1-CLEC5A-DsRED-Low Cell Line

64228

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Background of THP-1-CLEC5A-DsRED-Low Cell Line

CLEC5A (C-Type Lectin Domain Containing 5A) is a Protein Coding gene. This gene 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 signalling, glycoprotein turnover, and roles in inflammation and immune response. The encoded type II transmembrane protein interacts with dnax-activation protein 12 and may play a role in cell activation. Alternative splice variants have been described but their full-length sequence has not been determined.

Specifications

Catalog Number	KC-6559
Cell Line Name	THP-1-CLEC5A-DsRED-Low Cell Line
NCBI/UniProt Accession Number	NM_013252.3
Clone Number	16#
Host Cell Line	THP-1
Description	Stable THP-1 cell line expressing exogenous CLEC5A-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% basal medium+20% FBS+10% DMSO
Propagation Medium	RPMI 1640+10% FBS +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:6-1:8 every 2-3 days
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

THP-1-CLEC5A-DsRED-Low cell line was generated using a lentiviral vector expressing the CLEC5A-DsRED 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 1:6-1:8 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 cell 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

1.Lin J, Ou H, Wang W, Ma J, Zhang W, Liu Q. [Overexpression of CLEC5A inhibits cell proliferation and metastasis and reverses epithelial-mesenchymal transition in hepatocellular carcinoma]. Nan Fang Yi Ke Da Xue Xue Bao. 2023 Jan 20;43(1):85-91. Chinese.
2.Xavier-Carvalho C, Cezar RDDS, Freire NM, Vasconcelos CMM, Solorzano VEF, de Toledo-Pinto TG, Fialho LG, do Carmo RF, Vasconcelos LRS, Cordeiro MT, Baptista P, de Azeredo EL, da Cunha RV, de Souza LJ, Pacheco AG, Kubelka CF, Moura PMMF, Moraes MO. Association of rs1285933 single nucleotide polymorphism in CLEC5A gene with dengue severity and its functional effects. Hum Immunol. 2017 Oct;78(10):649-656.

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.