B-hB7-H3 MC38

Basic Information

Targeting strategy

The exogenous promoter and human B7-H3 coding sequence were inserted to replace part of murine exon 3 and all of exon 4. The insertion disrupts the endogenous murine B7h3 gene, resulting in a non-functional transcript.

Protein expression analysis

B7-H3 expression analysis in B-hB7-H3 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hB7-H3 MC38 cultures were stained with species-specific anti-B7-H3 antibody. Human B7-H3 was detected on the surface of B-hB7-H3 MC38 cells but not wild-type MC38 cells. The 2-F02 clone of B-hB7-H3 MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hB7-H3 MC38 cells. B-hB7-H3 MC38 cells (1×10⁶) and wild-type MC38 cells (5×10⁵) were subcutaneously implanted into C57BL/6 mice (female, 8-week-old, n=5). 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-hB7-H3 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Recommendations for inoculation in other mouse strains: 

1. Cell inoculation amount is recommended to be tried between 1×10⁶-1×10⁷; 
2. Inoculated cells can be tried to be suspended with DMEM stock solution.

Subcutaneous homograft tumor growth of B-hB7-H3 MC38 cells. B-hB7-H3 MC38 cells and wild-type MC38 cells were subcutaneously implanted into B-h4-1BB mice (female, 8-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B)  Body weight. Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hB7-H3 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

Protein expression analysis of tumor cells

B-hB7-H3 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human B7-H3 expression by flow cytometry. As shown, human B7-H3 was highly expressed on the surface of tumor cells. Therefore, B-hB7-H3 MC38 cells can be used for in vivo efficacy studies of novel B7-H3 therapeutics.

FMO Control 1: MC38 tumor cells only stained anti-mB7-H3 antibody; 

FMO Control 2: B-hB7-H3 MC38 tumor cells only stained anti-mB7-H3 antibody.