Basic Information
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Application
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B-hCD24 MC38 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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The exogenous promoter and human CD24 coding sequence was inserted to replace part of murine exon 1 and all of exon 2. The insertion disrupts the endogenous murine Cd24 gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
CD24 expression analysis in B-hCD24 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCD24 MC38 cultures were stained with an anti-human CD24 antibody. Human CD24 was detected on the surface of B-hCD24 MC38 cells but not wild-type MC38 cells. The 1-F02 clone of B-hCD24 MC38 cells was used for in vivo experiments.
Post-inoculation
B-hCD24 MC38 cells were subcutaneously implanted into C57BL/6 mice (n=5). Tumor cells were harvested on day 34 post-inoculation and assessed for CD24 expression by flow cytometry using species-specific antibodies. As shown, human CD24 was highly expressed on the surface of humanized B-hCD24 MC38 tumor cells, indicating B-hCD24 MC38 cells can be used for in vivo efficacy studies of novel CD24 therapeutics.
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Tumor Growth Curve and Body Weight Changes
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Tumor Growth Curve and Body Weight Changes in C57BL/6 Mice
Subcutaneous homograft tumor growth of B-hCD24 MC38 cells. B-hCD24 MC38 cells (1×106) and wild-type MC38 cells (1×106) were subcutaneously implanted into C57BL/6 mice (female, 6-7-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 × long diameter × short diameter2. As shown in panel A, B-hCD24 MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
