The mouse Her2 gene was replaced by human HER2 coding sequence in B-hHER2 MC38 cells. Human HER2 is highly expressed on the surface of B-hHER2 MC38 cells.

Gene targeting strategy for B-hHER2 MC38 cells. The exogenous CAG promoter and human HER2 coding sequence were inserted to replace part of murine exon 2 and all of exons 3-7. The insertion disrupts the endogenous murine Her2 gene, resulting in a non-functional transcript.

HER2 expression analysis in B-hHER2 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hHER2 MC38 cultures were stained with species-specific anti-HER2 antibody. Mouse HER2 was detectable in wild-type MC38 cells. Human HER2 was detected on the surface of B-hHER2 MC38 cells, but not on the surface of wild-type MC38 cells. The 2-B06 clone of B-hHER2 MC38 cells was used for in vivo tumor growth assays.

B-hHER2 MC38 cells were subcutaneously transplanted into B-hCD3E mice (n=5), and on 35 days post inoculation, tumor cells were harvested and assessed for human HER2 expression by flow cytometry. As shown, human HER2 was highly expressed on the surface of tumor cells. Therefore, B-hHER2 MC38 cells can be used for in vivo efficacy studies of HER2 therapeutics.

Subcutaneous homograft tumor growth of B-hHER2 MC38 cells. B-hHER2 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 7-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-hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

B-hHER2 MC38 tumor cells growth of individual mice. B-hHER2 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old, n=5). As shown in panel, B-hHER2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hHER2 MC38 cells.B-hHER2 MC38 cells (5x10⁵, 1x10⁶, 5x10⁶) were subcutaneously implanted into homozygous B-hCD3E mice (female, 7-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-hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hHER2 MC38 cells. B-hHER2 MC38 cells (5x10⁵, 1x10⁶, 5x10⁶) were subcutaneously implanted into homozygous B-h4-1BB mice (female, 7-week-old, n=6). 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-hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hHER2 MC38 cells. B-hHER2 MC38 cells (5x10⁵, 1x10⁶, 5x10⁶, 1x10⁷) were subcutaneously implanted into homozygous B-h4-1BB/hHER2 mice (female, 11-week-old, n=6). 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-hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hHER2 MC38 cells. B-hHER2 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into heterozygous B-hCD3E/hHER2 mice (female, 7-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-hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

B-hHER2 MC38 tumor cells growth of individual mice. B-hHER2 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into heterozygous B-hCD3E/hHER2 mice (female, 7-week-old, n=5). As shown in panel, B-hHER2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hHER2 MC38 tumor cells growth of individual mice. B-hHER2 MC38 cells (5x10⁵, 1x10⁶, 5x10⁶) were subcutaneously implanted into homozygous B-hCD3E mice (female, 7-week-old, n=5). As shown in panel, B-hHER2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hHER2 MC38 tumor cells growth of individual mice. B-hHER2 MC38 cells (5x10⁵, 1x10⁶, 5x10⁶) were subcutaneously implanted into homozygous B-h4-1BB mice (female, 7-week-old, n=6). As shown in panel, B-hHER2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hHER2 MC38 tumor cells growth of individual mice. B-hHER2 MC38 cells (5x10⁵, 1x10⁶, ⁵x10⁶, 1x10⁷) were subcutaneously implanted into homozygous B-h4-1BB/hHER2 mice (female, 11-week-old, n=6). As shown in panel, B-hHER2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.