Gene targeting strategy for B-hCD10 MC38 cells. The exogenous promoter and human CD10 coding sequence was inserted to replace part of murine exon 2-3. The insertion disrupts the endogenous murine CD10 gene, resulting in a non-functional transcript.

CD10 expression analysis in B-hCD10 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCD10 MC38 cultures were stained with species-specific anti-CD10 antibody. Human CD10 was detected on the surface of B-hCD10 MC38 cells but not wild-type MC38 cells. The 1-G07 clone of B-hCD10 MC38 cells was used for in vivo experiments.

Subcutaneous homograft tumor growth of B-hCD10 MC38 cells. B-hCD10 MC38 cells (5x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 11-12-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-hCD10 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hCD10 MC38 tumor cells growth of individual mice. B-hCD10 MC38 cells (5x106) and wild-type MC38 cells (5x105) were subcutaneously implanted into C57BL/6 mice (female, 11-12-week-old, n=6). As shown in panel, B-hCD10 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hCD10 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6), and on 28 days post inoculation, tumor cells were harvested and assessed for human CD10 expression by flow cytometry. As shown, human CD10 was highly expressed on the surface of tumor cells. Therefore, B-hCD10 MC38 cells can be used for in vivo efficacy studies of novel CD10 therapeutics.