The mouse Cd36 gene was replaced by human CD36 coding sequence in B-hCD36 MC38 cells. Human CD36 is highly expressed on the surface of B-hCD36 MC38 cells.

Gene targeting strategy for B-hCD36 MC38 cells. The exogenous promoter and human CD36 coding sequence were inserted to replace part of mouse Cd36. The insertion disrupts the endogenous murine Cd36 gene, resulting in a non-functional transcript.

CD36 expression analysis in B-hCD36 MC38 cells by flow cytometry. Single cell suspensions from B-hCD36 MC38 cultures were stained with species-specific anti-CD36 antibody. Human CD36 was detected on the surface of B-hCD36 MC38 cells but not wild-type MC38 cells. The 10-G12 clone of B-hCD36 MC38 cells was used for in vivo experiments.

Subcutaneous homograft tumor growth of B-hCD36 MC38 cells. B-hCD36 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into B-hCD36 mice (female, 7-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 × long diameter × short diameter². As shown in panel A, B-hCD36 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hCD36 MC38 cells were subcutaneously transplanted into B-hCD36 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human CD36 expression by flow cytometry. As shown, human CD36 was highly expressed on the surface of tumor cells. Therefore, B-hCD36 MC38 cells can be used for in vivo efficacy studies of novel CD36 therapeutics.