Basic Information
-
Application
-
B-hTPBG B16-F10 cells have the capability to establish tumors in vivo, which can be used for efficacy studies.
-
Targeting Strategy
-
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
-
Pre-inoculation
TPBG expression analysis in B-hTPBG B16-F10 cells by flow cytometry. Single cell suspensions from B-hTPBG B16-F10 cultures were stained with an anti-human 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.
Post-inoculation
B-hTPBG B16-F10 cells were subcutaneously transplanted into C57BL/6 mice (n=5). Tumor cells were harvested on day 19 post inoculation and assessed for human TPBG expression by flow cytometry. As shown, human TPBG was highly expressed on the surface of humanized B-hTPBG B16-F10 tumor cells, indicating B-hTPBG B16-F10 cells can be used for in vivo efficacy studies of novel TPBG therapeutics.
B-hTPBG B16-F10 cells were subcutaneously implanted into B-hCD40 mice and B-hPD-1/hCD40 mice (n=7). Tumor cells were harvested on day 14 post inoculation and assessed for human TPBG expression by flow cytometry. As shown, human TPBG was highly expressed on the surface of humanized B-hTPBG B16-F10 tumor cells, indicating B-hTPBG B16-F10 cells can be used for in vivo efficacy studies of novel TPBG therapeutics.
-
Tumor Growth Curve & Body Weight Changes
-
Tumor Growth Curve & Body Weight Changes in C57BL/6 Mice
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). (A) Average tumor volume ± SEM and (B) body weight (Mean± SEM) were measured twice a week. 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, which can be used for efficacy studies.
Tumor Growth Curve & Body Weight Changes in Humanized Mice
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). (A) Average tumor volume ± SEM and (B) body weight (Mean± SEM) were measured twice a week. 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, which can be used for efficacy studies.
