The mouse ENPP3 gene was replaced by human ENPP3 coding sequence in B-hENPP3 MC38 cells. Human ENPP3 is highly expressed on the surface of B-hENPP3 MC38 cells.

ENPP3 expression analysis in B-hENPP3 MC38 cells by flow cytometry. 

Single-cell suspensions from B-hENPP3 MC38 cultures were stained with species-specific anti-ENPP3 antibody. Human ENPP3 was detected on the surface of B-hENPP3 MC38 cells. The 2-A12 clone of B-hENPP3 MC38 cells was used for invivo experiments.

Subcutaneous homograft tumor growth of B-hENPP3 MC38 cells. 

B-hENPP3 MC38 cells (5x105) were subcutaneously implanted into C57BL/6JNifdc mice andB-hENPP3 mice (female, 6-7 weeks old, n=8). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight(Mean±SEM). (C) B-hENPP3 MC38 tumor growth of individual mice. Volume was expressed in mm3 using the formula: V=0.5 × long diameter × short diameter2.As shown in panel A, B-hENPP3 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

ENPP3 expression evaluated on B-hENPP3 MC38 tumor cells by flow cytometry. 

B-hENPP3 MC38 cells were subcutaneously transplanted into C57BL/6JNifdcmice and B-hENPP3 mice (female, 6-7 weeks old, n=8). At the end of the experiment, tumor cells were harvested and assessed for human ENPP3 expression by flowcytometry. As shown, human ENPP3 was highly expressed on the surface of tumor cells transplanted into B-hENPP3 mice. Therefore, B-hENPP3 MC38 cells can beused for in vivo efficacy studies of novel ENPP3 therapeutics.