B-hPD-L1 plus/hCEA MC38_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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B-hPD-L1 plus/hCEA MC38

Common name	B-hPD-L1 plus/hCEA MC38	Catalog number	320722
Aliases	PD-L1: CD274, PDCD1L1 CEA: CD66e, CEACAM5	Disease	Colon carcinoma
Organism	Mouse	Strain	C57BL/6
Tissue types	Colon	Tissue	Colon

Description

The mouse Pdl1 gene and Ceacam5 gene were replaced by human PD-L1 and CEACAM5 coding sequence in B-hPD-L1 plus/hCEA MC38 cells. Human PD-L1 and CEACAM5 are highly expressed on the surface of B-hPD-L1 plus/hCEA MC38 cells.

Application

B-hPD-L1 plus/hCEA MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.

Targeting strategy

Gene targeting strategy for B-hPD-L1 plus/hCEA 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 promoter and human CEACAM5 coding sequence was inserted into the ROSA26 locus in murine.

Protein expression analysis

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PD-L1 and CEACAM5 expression analysis in B-hPD-L1 plus/hCEA MC38 cells by flow cytometry. Single cell suspensions from MC38 and B-hPD-L1 plus/hCEA MC38 cultures were stained with species-specific anti-PD-L1 and anti-CEACAM5 antibodies. Human PD-L1 was detected on the surface of B-hPD-L1 plus/hCEA MC38 cells. Human CEACAM5 was detected on the surface of B-hPD-L1 plus/hCEA MC38 cells. The 2-D05 clone of B-hPD-L1 plus/hCEA MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

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Subcutaneous homograft tumor growth of B-hPD-L1 plus/hCEA MC38 cells. B-hPD-L1 plus/hCEA MC38 cells (5x10⁵) and B-hPD-L1 MC38 plus cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 5-8-week-old, n=5). 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 plus/hCEA MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.