Basic Information
Description
The expression cassettes containing exogenous promoter and human HER2 chemic CDS and the expression cassettes containing exogenous promoter and human TROP2 CDS were randomly inserted into the genome of B-Tg(hHER2/hTROP2) MC38 cells. Human HER2 and TROP2 are highly expressed on the surface of B-Tg(hHER2/hTROP2) MC38 cells.
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Targeting strategy
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Gene targeting strategy for B-Tg(hHER2/hTROP2) MC38 cells.
The expression cassettes containing exogenous promoter and human HER2 chemic CDS and the expression cassettes containing exogenous promoter and human TROP2 CDS were randomly inserted into the genome of B-Tg(hHER2/hTROP2) MC38 cells.
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Protein expression analysis
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hHER2/hTROP2 expression analysis in B-Tg(hHER2/hTROP2) MC38 cells by flow cytometry.
Single cell suspensions from wild-type MC38 and B-Tg(hHER2/hTROP2) MC38 cultures were stained with species-specific anti-HER2 and anti-TROP2 antibody. Human HER2 and TROP2 were detected on the surface of B-Tg(hHER2/hTROP2) MC38 cells but not wild-type MC38 cells. The MIX, 1-C07 and 1-D06 clone of B-Tg(hHER2/hTROP2) MC38 cells were used for in vivo tumor growth assays.
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Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-Tg(hHER2/hTROP2) MC38 cells.
B-Tg(hHER2/hTROP2) MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into heterozygous B-hHER2/hTROP2 mice (female, 27-week-old, n=10). 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-Tg(hHER2/hTROP2) MC38 cells were able to form tumors in vivo and can be used for efficacy studies.
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Protein expression analysis of tumor cells
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B-Tg(hHER2/hTROP2) MC38 cells were subcutaneously transplanted into heterozygous B-hHER2/hTROP2 mice (female, 27-week-old, n=10), and on 42 days post inoculation, tumor cells were harvested and assessed for human TROP2 expression by flow cytometry. As shown, human TROP2 was highly expressed on the surface of tumor cells. Therefore, B-Tg(hHER2/hTROP2) MC38 cells can be used for in vivo efficacy studies of HER2/TROP2 therapeutics.
B-Tg(hHER2/hTROP2) MC38 cells were subcutaneously transplanted into heterozygous B-hHER2/hTROP2 mice (female, 27-week-old, n=10), and on 42 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-Tg(hHER2/hTROP2) MC38 cells can be used for in vivo efficacy studies of HER2/TROP2 therapeutics.
