Basic Information
Description
The mouse Pd-l1 gene was replaced by the coding sequences of human PD-L1 and luciferase in B-hPD-L1 MC38 plus cells. Human PD-L1 is highly expressed on the surface of B-hPD-L1 MC38 plus cells. B-hPD-L1 MC38 plus cells have the capability to establish tumors in vivo and can be used for efficacy studies.
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Gene targeting strategy
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Gene targeting strategy for B-hPD-L1 MC38 plus cells. The exogenous promoter, human PD-L1 and luciferase coding sequences were inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Pd-l1 gene, resulting in a non-functional transcript.
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Protein expression analysis
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PD-L1 expression analysis in B-hPD-L1 MC38 plus cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hPD-L1 MC38 plus cultures were stained with species-specific anti-PD-L1 antibody. Human PD-L1 was detected on the surface of B-hPD-L1 MC38 plus cells but not wild-type MC38 cells. The 42 clone of B-hPD-L1 MC38 plus cells was used for in vivo experiments.
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Tumor growth curve
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Subcutaneous homograft tumor growth of B-hPD-L1 MC38 plus cells. B-hPD-L1 MC38 plus cells (2×106 and 5×105) were subcutaneously implanted into C57BL/6 mice (female, n=4). 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 MC38 plus cells were able to establish tumors in vivo and can be used for efficacy studies.
Subcutaneous homograft tumor growth of B-hPD-L1 MC38 plus cells. B-hPD-L1 MC38 plus cells (5×105,1×106 and 5×106) were subcutaneously implanted into C57BL/6J mice and homozygous B-hTIM3 mice. 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 MC38 plus were able to establish tumors in vivo and can be used for efficacy studies.