KC-2787

ID8-EPCAM-Luc2-Cell-Line

26799

Home » ID8-EPCAM-Luc2-Cell-Line

Background of ID8-EPCAM-Luc2-Cell-Line

Epithelial cell adhesion molecule (EpCAM/CD326) is a conserved type I transmembrane glycoprotein of 35 kDa expressed on epithelial cells of higher eukaryotes. Because EpCAM expression is limited to normal and malignant epithelia, it has been used as diagnostic marker for the detection of carcinoma cells in mesenchymal organs such as blood, bone marrow or lymph nodes.

Specifications

Catalog Number	KC-2787
Cell Line Name	ID8-EPCAM-Luc2-Cell-Line
Host Cell Line	ID8
Description	Stable ID8 clone expressing exogenous EPCAM and 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	70% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS +2μg/mL Puromycin+ 1000μg/mL Hygromycin
Selection Marker	Puromycin, hygromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:4 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

ID8-EPCAM-Luc2 cell line was generated using a lentiviral vector expressing the EPCAM and Luciferase sequence.

Characterization

Figure 1: Characterization of EPCAM-Luc2 overexpression in the ID8-EPCAM-Luc2 stable clone using FACS.

Figure 2: Characterization of luciferase overexpression in the ID8-EPCAM-Luc2 stable clone using Bright-Lite Luciferase System in the conditions of different cell number.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 2μg/mL puromycin and 1000μg/mL hygromycin)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 1:3-1:4 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

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.