Basic Information
Description
The mouse Cd19 gene was replaced by human CD19 coding sequence in B-hCD19-luc GFP A20 cells. Human CD19 is highly expressed on the surface this cell. And this cell expresses firefly luciferase as a marker.
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Targeting strategy
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Gene targeting strategy for B-hCD19-luc GFP A20 cells. The exogenous promoter, human CD19 coding sequence and luciferase was inserted to replace part of murine Cd19 gene in B-hCD19-luc GFP A20 cells. Mouse Cd19 transcript was disrupted, the protein no longer be expressed.
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Protein expression analysis
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CD19 expression analysis in B-hCD19-luc GFP A20 cells by flow cytometry. Single cell suspensions from wild-type A20 and B-hCD19-luc GFP A20 #2-C02 were stained with anti-CD19 antibody (BioLegend, anti-hCD19 antibody Cat.302212, anti-mCD19 antibody Cat.15555). Human CD19 was detected on the surface of B-hCD19-luc GFP A20 cells but not wild-type A20 cells, Mouse CD19 was not detected on B-hCD19-luc GFP A20 cells.
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In vitro luciferase activity of B-hCD19-luc GFP A20 cells
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Luminescence signal intensity of B-hCD19-luc GFP A20 cells. Cell lysates of wild-type A20 and B-hCD19-luc GFP A20 cells #2-C02 were measured using the Bright-GloTM luciferase Assay (Promega, Catalog No. E4030). B-hCD19-luc GFP A20 cells have a strong luminescence signal that is not present in wild-type A20 cells. -
Quantitative imaging data and body weight changes
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Growth kinetics of B-hCD19-luc GFP A20 tumors determined by bioluminescence imaging (BLI). B-hCD19-luc GFP A20 cells (5×105) were injected into the tail vein of wild-type BALB/c mice (female, 9-week-old, n=8). Signal intensity and body weight were measured twice a week. (A) Signal intensity. (B) Body weight. (C) Raw bioluminescence images. These results indicate that B-hCD19-luc GFP A20 cells can be used for in vivo efficacy evaluation. Values are expressed as mean ± SEM.
