KC-3762

BaF3-CLEC5A Cell Line

34732

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Background of BaF3-CLEC5A 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-3762
Cell Line Name	BaF3-CLEC5A Cell Line
Clone Number	3#
Host Cell Line	Ba/F3
Description	Ba/F3 cell line stably expressing exogenous CLEC5A 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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+1µg/mL Puromycin+8ng/mL mouse IL-3
Selection Marker	Puromycin
Morphology	Mostly single, round (some polymorph) cells in suspension
Subculture	Split saturated culture 1:10 every 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

BaF3-CLEC5A cell line was generated using a retrovirus vector expressing CLEC5A sequence.

Characterization

Figure 1: Characterization of CLEC5A overexpression in the BaF3 stable clone using FACS.

Figure2: Characterization of BaF3-CLEC5A cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10%FBS+1µg/mL Puromycin+8ng/mL mouse IL-3) 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 1:10 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 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

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. doi: 10.12122/j.issn.1673-4254.2023.01.11. PMID: 36856214; PMCID: PMC9978715.
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. doi: 10.1016/j.humimm.2017.07.013. Epub 2017 Jul 29. PMID: 28764923.