KC-4054

293T-human-COLEC12-Cell-Line

41865

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Background of 293T-human-COLEC12-Cell-Line

COLEC12 is a transmembrane scavenger receptor C-type lectin that is aberrantly expressed in a variety of cancers. collin subfamily member 12 May also play a role in inhibiting tumor immune response in gastric cancer, which can make it a new predictive marker and clinical therapeutic target for gastric cancer.

Specifications

Catalog Number	KC-4054
Cell Line Name	293T-human-COLEC12-Cell-Line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous human COLEC12 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 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

293T human COLEC12 cell line was generated using a lentiviral vector expressing the human COLEC12 sequence.

Characterization

Figure 1: Characterization of human COLEC12 overexpression in the 293T human COLEC12 stable clone using qPCR.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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 9.0mL 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 1:4-1:8 every 2-3 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

Sun X, Zhang Q, Shu P, Lin X, Gao X, Shen K. COLEC12 Promotes Tumor Progression and Is Correlated With Poor Prognosis in Gastric Cancer. Technol Cancer Res Treat. 2023 Jan-Dec;22:15330338231218163. doi: 10.1177/15330338231218163. PMID: 38112409; PMCID: PMC10734338.
Li GZ, Deng JF, Qi YZ, Liu R, Liu ZX. COLEC12 regulates apoptosis of osteosarcoma through Toll-like receptor 4-activated inflammation. J Clin Lab Anal. 2020 Nov;34(11):e23469. doi: 10.1002/jcla.23469. Epub 2020 Aug 21. PMID: 32822099; PMCID: PMC7676208.
Sun X, Zhang Q, Shu P, Lin X, Gao X, Shen K. COLEC12 Promotes Tumor Progression and Is Correlated With Poor Prognosis in Gastric Cancer. Technol Cancer Res Treat. 2023 Jan-Dec;22:15330338231218163. doi: 10.1177/15330338231218163. PMID: 38112409; PMCID: PMC10734338.