B-hEPCAM EL4

Basic Information

Description

The mouse Epcam gene was replaced by human EPCAM coding sequence in B-hEPCAM EL4 cells. Human EPCAM is highly expressed on the surface of B-hEPCAM EL4 cells.

Targeting strategy

Gene targeting strategy for B-hEPCAM EL4 cells. The exogenous promoter and human EPCAM coding sequence was inserted to replace part of murine exon 4 and all of exons 5~7. The insertion disrupts the endogenous murine Epcam gene, resulting in a non-functional transcript.

Protein Expression Analysis

EPCAM expression analysis in B-hEPCAM EL4 cells by flow cytometry. Single cell suspensions from wild-type EL4 and B-hEPCAM EL4 cultures were stained with species-specific anti-EPCAM antibody. Human EPCAM was detected on the surface of B-hEPCAM EL4 cells but not wild-type EL4 cells. The 4-B04 clone of B-hEPCAM EL4 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hEPCAM EL4 cells. B-hEPCAM EL4 cells (2×10⁵) and wild-type EL4 cells (2×10⁵) were subcutaneously implanted into C57BL/6 mice (female, 6-9-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B)  Body weight (Mean± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hEPCAM EL4 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hEPCAM EL4 cells. B-hEPCAM EL4 cells and wild-type EL4 cells were subcutaneously implanted into B-h4-1BB mice (female, 7-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. As shown in panel A, B-hEPCAM EL4 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

Protein expression analysis of tumor cells

B-hEPCAM EL4  cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human EPCAM expression by flow cytometry. As shown, human EPCAM was highly expressed on the surface of tumor cells. Therefore, B-hEPCAM EL4  cells can be used for in vivo efficacy studies of novel EPCAM therapeutics.