Expression cassettes containing the exogenous promoter and human TROP2 CDS were randomly inserted into the genome of B-hTROP2 MC38 plus cells. Human TROP2 was detected to be highly expressed on the surface of B-hTROP2 MC38 plus cells.
Expression cassettes containing the exogenous promoter and human TROP2 CDS were randomly inserted into the genome of MC38 cells that do not express the mouse TROP2 gene themselves.
Protein Expression Analysis
Human TROP2 expression analysis in B-hTROP2 MC38 plus cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hTROP2 MC38 plus cultures were stained with an anti-human TROP2 antibody. Human TROP2 was detected on the surface of B-hTROP2 MC38 plus cells but not wild-type MC38 cells. The 2-B10 clone of B-hTROP2 MC38 plus cells was used for in vivo tumor growth assays.
B-hTROP2 MC38 plus cells were subcutaneously transplanted into C57BL/6 mice (n=5).Tumor cells were harvested on day 28 post inoculation and assessed for species-specific TROP2 expression by flow cytometry. As shown, human TROP2 was highly expressed on the surface of humanized B-hTROP2 MC38 tumor cells, indicating B-hTROP2 MC38 plus cells can be used for in vivo efficacy studies of TROP2 therapeutics.
Tumor Growth Curve
Tumor Growth Curve & Body Weight Changes in C57BL/6 Mice
Subcutaneous homograft tumor growth of B-hTROP2 MC38 plus cells. B-hTROP2 MC38 plus cells (5×105) and wild-type MC38 cells (5×105) 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-hTROP2 MC38 plus cells were able to form tumors in vivo, which can be used for efficacy studies.