The mouse Mmp8 gene was replaced by human MMP-8 coding sequence in B-hMMP-8 MC38 cells. Human MMP-8 is highly expressed on the supernatant of B-hMMP-8 MC38 cells.

Gene targeting strategy for B-hMMP-8 MC38 cells. The exogenous promoter and human MMP-8 coding sequence was inserted to replace part of murine exons 1-3. The insertion disrupts the endogenous murine Mmp8 gene, resulting in a non-functional transcript.

MMP-8 expression analysis in B-hMMP-8 MC38 cells by western blot. Human MMP-8 was detected in the supernatant of B-hMMP-8 MC38 cells but not wild-type MC38 cells. The 1-D01 and 1-G12 clones of B-hMMP-8 MC38 cells was used for in vivo experiments.

Subcutaneous homograft tumor growth of B-hMMP-8 MC38 cells. B-hMMP-8 MC38 cells (5x106) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 12-13-week-old, n=6). 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-hMMP-8 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.