KC-2885

293T-Muc1 Cell Line

26815

Home » 293T-Muc1 Cell Line

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	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% 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.

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.