B-CAG-hPD-L1(v2)/hCEA MC38

Basic Information

Strain Name
MC38 Ceacam5tm1(hCEACAM5)Cd274tm2(hCD274)/Bcgen
Stock Number
320722
Common Name
B-CAG-hPD-L1(v2)/hCEA MC38
Disease
Colon Carcinoma
Species
Mus musculus, mouse
Growth factors
Adherent
Culture medium
DMEM+10%FBS+1%L-Glutamine+1%NEAA+1%Sodium pyruvate+1%HEPES
Frozen
90% FBS+10% DMSO

Targeting Strategy

Gene targeting strategy for B-CAG-hPD-L1(v2)/hCEA MC38. The human full-length CEACAM5 CDS region was inserted in the Ceacam5 exon1 of wild-type MC38. The human full-length CD274 CDS region was inserted in the Cd274 exon3 of wild-type MC38.

Details

Protein expression analysis

The PD-L1 expression analysis in B-CAG-hPD-L1(v2)/hCEA MC38 cells by flow cytometry. Single cells were collected from B-CAG-hPD-L1(v2) MC38 and B-CAG-hPD-L1(v2)/hCEA MC38 cells, and analyzed by flow cytometry with species-specific anti-PD-L1 antibody. Human PD-L1 were detectable in B-CAG-hPD-L1(v2)/hCEA MC38 and B-CAG-hPD-L1(v2) MC38 cells.

Protein expression analysis

The CEACAM5 expression analysis in B-CAG-hPD-L1(v2)/hCEA MC38 cells by flow cytometry. Single cells were collected from B-CAG-hPD-L1(v2) MC38 and B-CAG-hPD-L1(v2)/hCEA MC38 cells, and analyzed by flow cytometry with species-specific anti-CEACAM5 antibody. Human CEACAM was exclusively detectable in B-CAG-hPD-L1(v2)/hCEA MC38 but not B-CAG-hPD-L1(v2) MC38 cells.

Tumor Growth Curve & Body Weight Changes

Subcutaneous xenograft tumor growth of B-CAG-hPD-L1(v2)/hCEA MC38 cells. B-CAG-hPD-L1(v2)/hCEA MC38 cells and B-CAG-hPD-L1(v2) MC38 cells (5×105) were subcutaneously implanted into the C57BL/6 mice (female, 5-8 week-old, n=5).Tumor size and mice body weight were measured twice a week. (A) Mice body weight (Mean± SEM), (B) Tumor average volume ± SEM .Volume was expressed in mm3 using the formula: V=0.5a X b2, where a and b were the long and short diameters of the tumor, respectively. As shown in panel A, B-CAG-hPD-L1(v2)/hCEA MC38 were able to establish tumor in vivo and can be used for efficacy study.

Protein expression analysis in tumor cells

B-CAG-hPD-L1(v2)/hCEA MC38 cells were transplanted into C57BL/6 mice, and on 31 days post inoculation, certain volumes of tumors as was taken to detect the expression level of the cells. As Shown,human PD-L1 and CEACAM5 were highly expressed on the surface of tumor cells. B-CAG-hPD-L1(v2)/hCEA MC38 can be used for in vivo efficacy validation.

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