Basic Information
Description
The mouse Cd19 gene was replaced by human CD19 coding sequence in B-hCD19 EL4 cells. Human CD19 is highly expressed on the surface of B-hCD19 EL4 cells.
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Targeting strategy
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Gene targeting strategy for B-hCD19 EL4 cells.
The exogenous promoter and human CD19 coding sequence was inserted to replace part of murine exons 2-12. The insertion disrupts the endogenous murine CD19 gene, resulting in a non-functional transcript.
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Protein expression analysis
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The CD19 expression analysis in B-hCD19 EL4 cells by flow cytometry.
Single cells were collected from wild-type EL4 and B-hCD19 EL4 cells, and analyzed by flow cytometry with species-specific anti-CD19 antibody. Human CD19 was detectable in B-hCD19 EL4 cells but not wild-type EL4 cells. The 2-D07 clone of B-hCD19 EL4 cells was used for in vivo experiments.
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Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-hCD19 EL4 cells.
B-hCD19 EL4 cells (2×105) and wild-type EL4 cells (2×105) were subcutaneously implanted into C57BL/6N mice (female, 6-8-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-hCD19 EL4 cells were able to establish tumors in vivo and can be used for efficacy studies. Animals with tumor volume over 3000mm3 in the control group were euthanized.
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Protein expression analysis of tumor cells
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B-hCD19 EL4 cells were subcutaneously transplanted into C57BL/6 mice (n=8), and on 25 days post inoculation, tumor cells were harvested and assessed for human CD19 expression by flow cytometry. As shown, human CD19 was highly expressed on the surface of tumor cells. Therefore, B-hCD19 EL4 cells can be used for in vivo efficacy studies of novel CD19 therapeutics.
