The mouse Vcam1 gene was replaced by human CD106 (VCAM1) coding sequence in B-hCD106 MC38 cells. Human CD106 is highly expressed on the surface of B-hCD106 MC38 cells.

Gene targeting strategy for B-hCD106 MC38 cells. The exogenous promoter and human VCAM1 coding sequence was inserted to replace part of murine exon 2. The insertion disrupts the endogenous murine Vcam1 gene, resulting in a non-functional transcript.

CD106 expression analysis in B-hCD106 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCD106 MC38 cultures were stained with species-specific anti-CD106 antibody. Human CD106 was detected on the surface of B-hCD106 MC38 cells but not wild-type MC38 cells. The 7-C02 clone of B-hCD106 MC38 cells was used for in vivo experiments.

Subcutaneous homograft tumor growth of B-hCD106 MC38 cells. B-hCD106 MC38 cells (5x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 7-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 × long diameter × short diameter². As shown in panel A, B-hCD106 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hCD106 MC38 tumor cells growth of individual mice. B-hCD106 MC38 cells (5x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old, n=6). As shown in panel, B-hCD106 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hCD106 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6). At the end of the experiment, tumor cells were harvested and assessed for human CD106 expression by flow cytometry. As shown, human CD106 was highly expressed on the surface of tumor cells. Therefore, B-hCD106 MC38 cells can be used for in vivo efficacy studies of novel CD106 therapeutics.