KC-2613

Pan02-TROP2 Cell Line

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Background

This intronless gene encodes a carcinoma-associated antigen highly expressed on various epithelial tumors and correlates with poor prognosis. This antigen is a cell surface receptor that transduces calcium signals. Mutations of this gene have been associated with gelatinous drop-like corneal dystrophy.

Specifications

Catalog Number:
KC-2613
Cell Line Name:
Pan02-TROP2 Cell Line
Host Cell Line:
Pan02
Description:
Stable Pan02 clone expressing exogenous human TROP2 gene
Quantity:
One vial of frozen cells (5×106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% 1640 + 20% FBS + 10% DMSO
Propagation Medium:
RPMI1640+10%FBS+2μg/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:
N/A
Mycoplasma Status:
Negative

Cell Line Generation

Pan02-TROP2 Cell Line was generated using a lentiviral vector expressing the TROP2 sequence.

Characterization

Figure 1: Characterization of TROP2 overexpression in the Pan02-TROP2 stable clone using FACS.

Application

Cell Resuscitation

1. Prewarm culture medium (1640 supplemented with 10% FBS and 2μg/mL puromycin)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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% 1640 + 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. Okajima D, Yasuda S, Maejima T, Karibe T, Sakurai K, Aida T, Toki T, Yamaguchi J, Kitamura M, Kamei R, Fujitani T, Honda T, Shibutani T, Muramatsu S, Nakada T, Goto R, Takahashi S, Yamaguchi M, Hamada H, Noguchi Y, Murakami M, Abe Y, Agatsuma T. Datopotamab Deruxtecan, a Novel TROP2-directed Antibody-drug Conjugate, Demonstrates Potent Antitumor Activity by Efficient Drug Delivery to Tumor Cells. Mol Cancer Ther. 2021 Dec;20(12):2329-2340. doi: 10.1158/1535-7163.MCT-21-0206. Epub 2021 Aug 19. PMID: 34413126; PMCID: PMC9398094.
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