KC-2885

293T-Muc1 Cell Line

26815

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Background of 293T-Muc1 Cell Line

MUC1 is a membrane mucin expressed on the apical surface of lung epithelial cells. MUC1 has anti-adhesion and immunosuppressive properties, prevents infection, and is involved in carcinogenic processes as well as cell signaling. Therefore, MUC1 has become a target for mouse and human tumor immunotherapy research.

Specifications

Catalog Number	KC-2885
Cell Line Name	293T-Muc1 Cell Line
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous 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 1:4-1:5 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-Muc1 cell line was generated using a lentiviral vector expressing the Muc1 sequence.

Characterization

Figure 1: Characterization of Muc1 overexpression in the 293T-Muc1 stable clone using FACS.

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 1:4-1:5 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

Kato K, Zemskova MA, Hanss AD, Kim MM, Summer R, Kim KC. Muc1 deficiency exacerbates pulmonary fibrosis in a mouse model of silicosis. Biochem Biophys Res Commun. 2017 Nov 25;493(3):1230-1235. doi: 10.1016/j.bbrc.2017.09.047. Epub 2017 Sep 12. PMID: 28916165.
Apostolopoulos V, Stojanovska L, Gargosky SE. MUC1 (CD227): a multi-tasked molecule. Cell Mol Life Sci. 2015 Dec;72(23):4475-500. doi: 10.1007/s00018-015-2014-z. Epub 2015 Aug 21. PMID: 26294353.
Kato K, Zemskova MA, Hanss AD, Kim MM, Summer R, Kim KC. Muc1 deficiency exacerbates pulmonary fibrosis in a mouse model of silicosis. Biochem Biophys Res Commun. 2017 Nov 25;493(3):1230-1235. doi: 10.1016/j.bbrc.2017.09.047. Epub 2017 Sep 12. PMID: 28916165.