B-hTPBG B16-F10

Basic Information

Targeting Strategy

Gene targeting strategy for B-hTPBG B16-F10 cells. The exogenous promoter and human TPBG coding sequence was inserted to replace part of murine exon 2. The insertion disrupts the endogenous murine Tpbg gene, resulting in a non-functional transcript.

Protein Expression Analysis

TPBG expression analysis in B-hTPBG B16-F10 cells by flow cytometry. Single cell suspensions from B-hTPBG B16-F10 cultures were stained with species-specific anti-TPBG antibody. Human TPBG was detected on the surface of B-hTPBG B16-F10 cells but not wild-type B16-F10 cells. The 2-E11 clone of B-hTPBG B16-F10 cells was used for in vivo experiments.

Tumor Growth Curve & Body Weight Changes

Subcutaneous homograft tumor growth of B-hTPBG B16-F10 cells. B-hTPBG B16-F10 cells (2×10⁵) and wild-type B16-F10 cells (2×10⁵) 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-hTPBG B16-F10 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hTPBG B16-F10 cells. B-hTPBG B16-F10 cells (2×10⁵) were subcutaneously implanted into B-hCD40 mice and B-hPD-1/hCD40 mice (female, 7-week-old, n=7). 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-hTPBG B16-F10 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hTPBG B16-F10 cells. B-hTPBG B16-F10 cells (1×10⁵, 2×10⁵, 5×10⁵) and wild-type B16-F10 cells (2×10⁵) were subcutaneously implanted into B-hCD3E mice (female, 7-9-week-old). 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-hTPBG B16-F10 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hTPBG B16-F10 cells. B-hTPBG B16-F10 cells and wild-type MC38 cells were subcutaneously implanted into B-h4-1BB mice (female, 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. As shown in panel A, B-hTPBG B16-F10 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-hTPBG B16-F10 cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human TPBG expression by flow cytometry. As shown, human TPBG was highly expressed on the surface of tumor cells. Therefore, B-hTPBG B16-F10 cells can be used for in vivo efficacy studies of novel TPBG therapeutics.

B-hTPBG B16-F10 cells were subcutaneously transplanted into B-hCD40 mice and B-hPD-1/hCD40 mice (n=7), and on 14 days post inoculation, tumor cells were harvested and assessed for human TPBG expression by flow cytometry. As shown, human TPBG was highly expressed on the surface of tumor cells. Therefore, B-hTPBG B16-F10 cells can be used for in vivo efficacy studies of novel TPBG therapeutics.