• Origin: The MC38 cell line is derived from C57BL6 murine colon adenocarcinoma cells. The cell line is a commonly used murine model for colorectal carcinoma.
• Background Information: TROP2 is a tumor-associated calcium ion signal transduction protein and a cell surface glycoprotein, which is highly expressed in tumor tissues. It can promote the growth, proliferation and metastasis of tumor cells by regulating the calcium signaling pathway, the expression of cyclin and reducing the adhesion of fibronectin.
• Gene targeting strategy: The exogenous promoter and human TROP2 CDS was inserted into the mouse Trop2 exon 1.
• Tumorigenicity: Confirmed in B-hTROP2 mice.
• Application: B-hTROP2 high MC38 tumor models can be used for preclinical evaluation of antibody drugs, ADC targeting human TROP2.
• Notes:

Inoculated cell lines can be suspended with DMEM stock solution.

Before implementing the project, it is recommended to perform tumor growth experiments. The recommended cell inoculation amount is between 5E5~1E6.

In the experiment, it is necessary to ensure that the number of animals inoculated subcutaneously is at least 1.6 times the actual grouping number.

• Human TROP2 was detected on the surface of B-hTROP2 high MC38 but not wild-type MC38 cells.

TROP2 expression analysis in B-hTROP2 high MC38 by flow cytometry. Single cell suspensions from wild-type MC38 and B-hTROP2 high MC38 #1-H02 cultures were stained with anti-TROP2 antibody (BioLegend, 363804).

Subcutaneous tumor growth of B-hTROP2 high MC38. B-hTROP2 high MC38 (1x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into homozygous B-hTROP2 mice (8-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². Results indicate that B-hTROP2 high MC38 were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

• Human TROP2 was detected on the surface of B-hTROP2 high MC38 tumors but not wild-type MC38 cells.

TROP2 expression evaluated on B-hTROP2 high MC38 cells by flow cytometry. B-hTROP2 high MC38 cells were subcutaneously transplanted into homozygous B-hTROP2 mice (n=6). Upon conclusion of the experiment, tumor cells were harvested and analyzed with anti-TROP2 antibody (BioLegend, 363804) by flow cytometry. Therefore, B-hTROP2 high MC38 cells can be used for in vivo efficacy studies evaluating novel TROP2 antibody drugs therapeutics.