Basic Information

Common name
B-hPD-L1 MC38 ad
Catalog number
321852
Aliases
DPPIVA, PD-L1alpha, SIMP
Disease
Colon carcinoma
Organism
Mouse
Strain
C57BL/6
Tissue types
Colon
Tissue
Colon
Application
B-hPD-L1 MC38 ad cells have the capability to establish tumors in vivo and can be used for efficacy studies.
NCBI Gene ID

Description

The exogenous promoter and 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 MC38 ad cells.

Targeting strategy

Gene targeting strategy for B-hPD-L1 MC38 ad cells. The exogenous promoter and 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 MC38 ad cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hPD-L1 MC38 ad cultures were stained with species-specific anti-PD-L1 antibody. Human PD-L1 was detected on the surface of B-hPD-L1 MC38 ad cells but not wild-type MC38 cells. The 2-A10 clone of B-hPD-L1 MC38 ad cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hPD-L1 MC38 ad cells. B-hPD-L1 MC38 ad cells (2×105), B-hPD-L1 MC38 ad cells (1×106) 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 MC38 ad cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

Tumor volume and weight measurements

Protein expression analysis of tumor cells

B-hPD-L1 MC38 ad cells were subcutaneously transplanted into C57BL/6 mice (n=5), and on 28 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 MC38 ad cells can be used for in vivo efficacy studies of novel PD-L1 therapeutics.