The mouse Itga7 gene was replaced by human ITGA7 coding sequence in B-hITGA7 MC38 cells. Human ITGA7 is highly expressed on the surface of B-hITGA7 MC38 cells.
 

Gene targeting strategy for B-hITGA7 MC38 cells. The exogenous promoter and human ITGA7 coding sequence was inserted to replace part of murine exon 1. The insertion disrupts the endogenous murine Itga7 gene, resulting in a non-functional transcript.

ITGA7 expression analysis in B-hITGA7 MC38 cells by flow cytometry and western blot. Single cell suspensions from wild-type MC38 and B-hITGA7 MC38 cultures were stained with species-specific anti-ITGA7 antibody. Mouse ITGA7 was not detected on the surface of B-hITGA7 MC38. Human ITGA7 was detectable in B-hITGA7 MC38 cells and wild-type MC38 cells, as the antibody is crossly reactive with ITGA7 in human and mice. The 1-D10 clone of B-hITGA7 MC38 cells was used for in vivo experiments.

Subcutaneous homograft tumor growth of B-hITGA7 MC38 cells. B-hITGA7 MC38 cells (5x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 8-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-hITGA7 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hITGA7 MC38 tumor cells growth of individual mice. B-hITGA7 MC38 cells (5x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 8-week-old, n=6). As shown in panel, B-hITGA7 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.