KC-2599-DW

RM1-Trop2 Cell Line

60541

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Background of RM1-Trop2 Cell Line

This intronless gene encodes a carcinoma-associated antigen. This antigen is a cell surface receptor that transduces calcium signals. Mutations of this gene have been associated with gelatinous drop-like corneal dystrophy. Trop2, as named as TACSTD2. Diseases associated with TACSTD2 include Corneal Dystrophy, Gelatinous Drop-Like and Corneal Dystrophy. Among its related pathways are Ca, cAMP and Lipid Signaling and Adhesion. Gene Ontology (GO) annotations related to this gene include signaling receptor activity. An important paralog of this gene is EPCAM.

Specifications

Catalog Number	KC-2599-DW
Cell Line Name	RM1-Trop2 Cell Line
NCBI/UniProt Accession Number	NM_002353.3
Clone Number	4#
Host Cell Line	RM1
Description	Stable RM1 cell line expressing exogenous Trop2 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:2-1:3 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

Cell Line Generation

RM1-Trop2 cell line was generated using a lentiviral vector expressing the Trop2 sequence.

Characterization

Figure 1: Characterization of Trop2 overexpression in RM1-Trop2 cell line stable clone using FACS.

Figure 2: Validation of in vivo tumorigenicity of RM1 cells stably expressing Trop2 via subcutaneous implantation in C57BL/6J mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.

Cell Resuscitation

Prewarm culture medium (RPMI1640 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 1:2-1:3 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Zhao P, Yu HZ, Cai JH. Clinical investigation of TROP-2 as an independent biomarker and potential therapeutic target in colon cancer. Mol Med Rep. 2015 Sep;12(3):4364-4369. doi: 10.3892/mmr.2015.3900. Epub 2015 Jun 8. PMID: 26059528.
Addati T, Achille G, Centrone M, Petroni S, Popescu O, Russo S, Grammatica L, Simone G. TROP-2 expression in papillary thyroid cancer: a preliminary cyto-histological study. Cytopathology. 2015 Oct;26(5):303-11. doi: 10.1111/cyt.12196. Epub 2014 Aug 27. PMID: 25164548.