Basic Information

Common Name
B-hPD-L1 low MC38
Aliases
CD274, PDCD1L1
Catalog Number
310889
Cell Type/ Disease
Colon Carcinoma
Species/ Strain
Mus musculus, C57BL/6
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.

Description

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

Targeting Strategy

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.

Protein expression analysis

PD-L1 expression analysis in B-hPD-L1 low MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38, B-hPD-L1 MC38 plus and B-hPD-L1 low MC38 cultures were stained with species-specific anti-PD-L1 antibody. Mouse PD-L1 was detectable in wild-type MC38 cells. Human PD-L1 was detectable in B-hPD-L1 low MC38 cells, and human PD-L1 was expressed highly on the surface of B-hPD-L1 MC38 plus cells. The 2-C09 clone of B-hPD-L1 low MC38 cells was used for in vivo experiments.

Tumor growth curve

Subcutaneous homograft tumor growth of B-hPD-L1 low MC38 cells. B-hPD-L1 low MC38 cells (1×106), B-hPD-L1 MC38 plus cells (5×105) and wild-type MC38 cells (1×106) were subcutaneously implanted into C57BL/6N mice (female, 7-week-old, n=5). Tumor volume and body weight were measured twice a week. 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 low MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. (A) Average tumor volume ± SEM. (B)  Body weight (Mean± SEM).

The subcutaneous tumor formation of B-hPD-L1 low MC38 cells was slow, and the inoculation dose was recommended to be increased to 1×106.