KC-2785

S180-EPCAM-Luc2 Cell Line

26795

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Background of S180-EPCAM-Luc2 Cell Line

Epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein functioned as cell-cell adhesion in epithelia, cell migration, proliferation and differentiation. EpCAM have oncogenic potential through upregulation of c-myc, cyclins A & E after cleavage. Epcam is not only used as diagnostic marker for various cancer, but also a potential target for cancer therapy. Luciferase is an oxidative enzyme that can produce bioluminescence with addition of luciferin, but don’t need an external light source unlike of fluorescent proteins. Photo emission can be detected directly by light sensitive device. Such as luminometer or modified microscope. Luciferase is widely used in many fields of biological research, such as transcriptional activity, kinase or other enzyme activity, cellular ATP level, whole animal imaging.

Specifications

Catalog Number	KC-2785
Cell Line Name	S180-EPCAM-Luc2 Cell Line
Host Cell Line	S180-EPCAM
Description	Stable S180-EPCAM clone expressing exogenous Luciferase 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	Frozen in liquid nitrogen with 70% RPMI1640, 20% FBS and 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 2μg/mL puromycin+300μg/mL hygromycin B
Selection Marker	Puromycin, Hygromycin B
Morphology	Suspension and adherence
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

S180-Epcam-Luc2 cell Line was generated using a retroviral vector expressing human Epcam and luciferase sequence.

Characterization

Figure 1: Characterization of Epcam overexpression in S180 stable clones.

Figure 2: Characterization of luciferase expression in S180 using the Bright-Glo Detection.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 2μg/mL puromycin+300μg/mL Hygromycin B)in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO₂ incubator. 8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

References

1. Litvinov, Sergey; et al. (1994). Ep-CAM: a human epithelial antigen is a homophilic cellÿcell adhesion molecule. The Journal of Cell Biology. 125 (2): 437ÿ46. 2. Maetzel, Dorothea; et al. (2009). Nuclear signalling by tumour-associated antigen EpCAM. Nature Cell Biology. 11 (2): 162ÿ71. 3. Osta, WA; et al. (2004). EpCAM is overexpressed in breast cancer and is a potential target for breast cancer gene therapy. Cancer Res. 64 (16): 5818ÿ24. 2. Gaber A, Lenarčič B, Pavšič M. Current View on EpCAM Structural Biology. Cells. 2020 May 31;9(6):1361. doi: 10.3390/cells9061361. PMID: 32486423; PMCID: PMC7349879.