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
Culture Properties
Adherent
Culture medium
DMEM+10%FBS+1%L-Glutamine+1%NEAA+1%Sodium pyruvate+1%HEPES
Frozen
90% FBS+10% DMSO
Notice
Biocytogen provides engineering service to generate the humanized tumor cell lines from customer’s own cell lines. Our humanized tumor cell lines are for pharmacology services ONLY.

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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