Basic Information
Description
The mouse B7-h3 gene was replaced by the human B7-H3 coding sequence in B-hB7-H3 CT26.WT cells. Human B7-H3 is highly expressed on the surface of B-hB7-H3 CT26.WT cells.
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Application
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B-hB7-H3 CT26.WT cells have the capability to establish tumors in vivo, which can be used for efficacy studies.
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Targeting strategy
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Gene targeting strategy for B-hB7-H3 CT26.WT cells. The chemic human and mouse B7-H3 coding sequence was inserted to replace part of murine exon 3 and exon 4. The insertion disrupts the endogenous murine B7-h3 gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
B7-H3 expression analysis in B-hB7-H3 CT26.WT cells by flow cytometry. Single cell suspensions from B-hB7-H3 CT26.WT cultures were stained with an anti-human B7-H3 antibody. Human B7-H3 was detected on the surface of B-hB7-H3 CT26.WT cells but not wild-type CT26.WT cells. The 3-H02 clone of B-hB7-H3 CT26.WT cells was used for in vivo experiments.
Post-inoculation
B-hB7-H3 CT26.WT cells were subcutaneously implanted into BALB/c mice (n=5). Tumor cells were harvested on day 34 post inoculation and assessed for human B7-H3 expression by flow cytometry. As shown, human B7-H3 was highly expressed on the surface of humanized B-hB7-H3 CT26.WT tumor cells, indicating B-hB7-H3 CT26.WT cells can be used for in vivo efficacy studies of novel B7-H3 therapeutics.
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Tumor Growth Curve
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Tumor Growth Curve & Body Weight Changes in BALB/c Mice
Subcutaneous homograft tumor growth of B-hB7-H3 CT26.WT cells. B-hB7-H3 CT26.WT cells and wild-type CT26.WT cells were subcutaneously implanted into BALB/c mice (female, 7-week-old, n=5). (A) Average tumor volume ± SEM and (B) body weight (mean ± SEM) 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-hB7-H3 CT26.WT cells were able to establish tumors in vivo, which can be used for efficacy studies.
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Protein expression analysis of tumor cells
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B-hB7-H3 CT26.WT cells were subcutaneously transplanted into BALB/c mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human B7-H3 expression by flow cytometry. As shown, human B7-H3 was highly expressed on the surface of tumor cells. B-hB7-H3 CT26.WT cells can be used for in vivo efficacy studies of novel B7-H3 therapeutics.
