The exogenous promoter and human PD-L1 CDS was inserted into the mouse Pd-l1 exon 3, the exogenous promoter and human CLDN18.2 CDS was inserted into mouse Rosa26 locus site in B-hPD-L1 plus/hCLDN18.2 MC38. Human PD-L1 and CLDN18.2 were detected on the surface of B-hPD-L1 plus/hCLDN18.2 MC38 cells.
Application
B-hPD-L1 plus/hCLDN18.2 MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.
Targeting strategy
Gene targeting strategy for B-hPD-L1 plus/hCLDN18.2 MC38. The exogenous promoter and human PD-L1 CDS was inserted into the mouse Pd-l1 exon 3, the exogenous promoter and human CLDN18.2 CDS was inserted into mouse Rosa26 locus site in B-hPD-L1 plus/hCLDN18.2 MC38.
Protein expression analysis
PD-L1 and CLDN18.2 expression analysis in B-hPD-L1 plus/hCLDN18.2 MC38 cells by flow cytometry. Single cell suspensions from B-hPD-L1 plus/hCLDN18.2 MC38 cultures were stained with anti-PD-L1 and anti-CLDN18.2 antibodies. Human PD-L1 and CLDN18.2 were detected on the surface of B-hPD-L1 plus/hCLDN18.2 MC38 cells.
Tumor growth curve & Body weight changes
Subcutaneous homograft tumor growth of B-hPD-L1 plus/hCLDN18.2 MC38 cells. B-hPD-L1 plus/hCLDN18.2 MC38 cells and wild-type MC38 cells were subcutaneously implanted into C57BL/6N mice (female, 10-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-hPD-L1 plus/hCLDN18.2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Tumor growth curve of individual mice
B-hPD-L1 plus/hCLDN18.2 MC38 tumor growth of individual mice. B-hPD-L1 plus/hCLDN18.2 MC38 cells and wild-type MC38 cells were subcutaneously implanted into C57BL/6N mice (female, 10-week-old, n=6). As shown in panel, B-hPD-L1 plus/hCLDN18.2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.