KC-2244

293T-cyno-Muc1-Cell-Line

23672

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Background of 293T-cyno-Muc1-Cell-Line

The mucin (MUC) family is a group of highly glycosylated macromolecules that are abundantly expressed in mammalian epithelial cells. Mucins contribute to the formation of the mucus barrier and are therefore protective against infection. Interestingly, some MUC proteins are aberrantly expressed in cancer cells and are involved in tumourigenesis and progression, including cell growth, proliferation, apoptosis inhibition, chemoresistance, metabolic reprogramming and immune evasion.

Specifications

Catalog Number	KC-2244
Cell Line Name	293T-cyno-Muc1-Cell-Line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous cyno Muc1 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 cyno Muc1 cell line was generated using a lentiviral vector expressing the cyno Muc1 sequence.

Characterization

Figure 1: Characterization of cyno Muc1 overexpression in the 293T cyno Muc1 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

Qing L, Li Q, Dong Z. MUC1: An emerging target in cancer treatment and diagnosis. Bull Cancer. 2022 Nov;109(11):1202-1216. doi: 10.1016/j.bulcan.2022.08.001. Epub 2022 Sep 30. PMID: 36184332.
Kufe DW. Emergence of MUC1 in Mammals for Adaptation of Barrier Epithelia. Cancers (Basel). 2022 Sep 30;14(19):4805. doi: 10.3390/cancers14194805. PMID: 36230728; PMCID: PMC9564314.
Kufe D. Dependence on MUC1-C in Progression of Neuroendocrine ProstateCancer. Int J Mol Sci. 2023 Feb 13;24(4):3719. doi: 10.3390/ijms24043719. PMID: 36835130; PMCID: PMC9967814.