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

Subcutaneous homograft tumor growth of B-hB7-H3 MC38 cells. B-hB7-H3 MC38 cells (5×10⁵) and wild-type MC38 cells (5×10⁵) were subcutaneously implanted into C57BL/6N mice (female, 5-8-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B)  Bodyweight (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 5×10⁵-1×10⁷;

2.Inoculated cells can be tried to be suspended with DMEM stock solution.