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B-hPD-L1 plus/hCD47 MC38
Common name
B-hPD-L1 plus/hCD47 MC38 Catalog number     320714
Aliases CD274, PDCD1L1
  IAP, MER6, OA3
Disease  Colon carcinoma
Organism
Mouse
Strain  C57BL/6
Tissue types Colon Tissue  Colon

Description 

The mouse Pdl1 gene and Cd47 gene were replaced by human PD-L1 and CD47 coding sequence in B-hPD-L1 plus/hCD47 MC38 cells. Human PD-L1 and CD47 are highly expressed on the surface of B-hPD-L1 plus/hCD47 MC38 cells.

Targeting strategy

Gene targeting strategy for B-hPD-L1 plus/hCD47 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 CD47 coding sequence was inserted to replace part of murine exon 2 and all of exon 3. The insertion disrupts the endogenous murine Cd47 gene, resulting in a non-functional transcript.

Protein expression analysis


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PD-L1 and CD47 expression analysis in B-hPD-L1 plus/hCD47 MC38 cells by flow cytometry. Single cell suspensions from B-hPD-L1 plus/hCD47 MC38 cultures were stained with species-specific anti-PD-L1 and anti-CD47 antibodies. Human PD-L1 and CD47 were detected on the surface of B-hPD-L1 plus/hCD47 MC38 cells. The 3-D07 clone of B-hPD-L1 plus/hCD47 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/hCD47 MC38 cells. B-hPD-L1 plus/hCD47 MC38 cells (5x105) were subcutaneously implanted into B-hPD-L1/hSIRPA/hCD47 mice (female, 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/hCD47 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.