Basic Information
Description
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.
-
Targeting strategy
-
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
-
Pre-inoculation
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.
Post-inoculation
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.
