Basic Information

Common name
B-hPD-L1/hHER2 MC38
Catalog number
321917
Aliases
CD274, B7-H, B7H1, PD-L1, PDCD1L1, PDCD1LG1, PDL1, hPD-L1, CD274 molecule, Programmed cell death 1 ligand 1; ERBB2, CD340, HER-2, HER-2/neu, HER2, MLN 19, NEU, NGL, TKR1, VSCN2, c-ERB-2, c-ERB2, erb-b2 receptor tyrosine kinase 2
Disease
Colon carcinoma
Organism
Mouse
Strain
C57BL/6
Tissue types
Colon
Tissue
Colon
Application
B-hPD-L1/hHER2 MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.
NCBI Gene ID

Description

The mouse Pdl1 gene was replaced by human PD-L1 coding sequence in B-hPD-L1/hHER2 MC38 cells. Human PD-L1 is highly expressed on the surface of B-hPD-L1/hHER2 MC38 cells.

The mouse Erbb2 gene was replaced by human ERBB2 coding sequence in B-hPD-L1/hHER2 MC38 cells. Human ERBB2 is highly expressed on the surface of B-hPD-L1/hHER2 MC38 cells.

Gene targeting strategy

Gene targeting strategy for B-hPD-L1/hHER2 MC38 cells. The exogenous promoter and human PD-L1 coding sequence was inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Pdl1 gene, resulting in a non-functional transcript.

The exogenous CAG promoter and human ERBB2 coding sequence was inserted to replace part of murine exon 2 and all of exons 3-7. The insertion disrupts the endogenous murine Erbb2 gene, resulting in a non-functional transcript.

Protein expression analysis

 

 

 

 

 

 

PD-L1 and HER2 protein expression analysis in B-hPD-L1/hHER2 MC38 cells. Single cell suspensions from wild-type MC38 and B-hPD-L1/hHER2 MC38 cultures were stained with species-specific anti-PD-L1 and anti-HER2 antibodies. Human PD-L1 and HER2 were detected on the surface of B-hPD-L1/hHER2 MC38 cells but not wild-type MC38 cells. The 3-B08 clone of B-hPD-L1/hHER2 MC38 cells was used for in vivo tumor growth assays.

Tumor growth curve in wild-type mice

Subcutaneous homograft tumor growth of B-hPD-L1/hHER2 MC38 cells. B-hPD-L1/hHER2 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 9-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/hHER2 MC38 cells were able to form tumors in vivo, which can be used for efficacy studies.

Tumor volume and weight measurements

Protein expression analysis post-inoculation

 

 

 

 

 

 

 

 

 

B-hPD-L1/hHER2 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6), 39 days post inoculation, tumor cells were harvested and assessed for human PD-L1 and HER2 protein expression by flow cytometry. As shown, human PD-L1 and HER2 were highly expressed on the surface of tumor cells. Therefore, B-hPD-L1/hHER2 MC38 cells can be used for in vivo efficacy studies to test novel PD-L1 and HER2 therapeutics.