Basic Information

Common Name
B-hPD-L1 low MC38 plus
Catalog number
321853
Aliases
B7-H, B7H1, PD-L1, PDCD1L1, PDCD1LG1, PDL1, hPD-L1
Disease
Colon carcinoma
Organism
Mus musculus, mouse
Strain
C57BL/6
Tissue types
Colon
Tissue
Colon
Application
B-hPD-L1 low MC38 plus cells have the capability to establish tumors in vivo and can be used for efficacy studies.
NCBI Gene ID

Description

The chimeric CDS composed of mouse signal peptide, human IgV region, mouse extracellular region except IgV domain, moue transmembrane region and mouse intracellular region of PD-L1 gene was inserted to replace the IgV domain of mouse Pdl1 gene. Human PD-L1 is highly expressed on the surface of B-hPD-L1 low MC38 plus cells.

Targeting strategy

Gene targeting strategy for B-hPD-L1 low MC38 plus cells. The chimeric CDS composed of mouse signal peptide, human IgV region, mouse extracellular region except IgV domain, moue transmembrane region and mouse intracellular region of PD-L1 gene was inserted to replace the IgV domain of mouse Pdl1 gene. The insertion disrupts the endogenous mouse Pdl1 gene, resulting in a non-functional transcript.

Protein expression analysis

PD-L1 expression analysis in B-hPD-L1 low MC38 plus cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hPD-L1 low MC38 plus cultures were stained with species-specific anti-PD-L1 antibody. Human PD-L1 was detected on the surface of B-hPD-L1 low MC38 plus cells but not wild-type MC38 cells. The 2-D10 clone of B-hPD-L1 low MC38 plus cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hPD-L1 low MC38 plus cells. B-hPD-L1 low MC38 plus cells (1×106), B-hPD-L1 low MC38 plus cells (3×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 9-week-old, n=5). 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-hPD-L1 low MC38 plus 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-hPD-L1 low MC38 plus cells were subcutaneously transplanted into C57BL/6 mice (n=5), and on 29 days post inoculation, tumor cells were harvested and assessed for human PD-L1 expression by flow cytometry. As shown, human PD-L1 was highly expressed on the surface of tumor cells. Therefore, B-hPD-L1 low MC38 plus cells can be used for in vivo efficacy studies of novel PD-L1 therapeutics.