Basic Information
Description
The mouse Cxcr5 gene was replaced by the human CXCR5 coding sequence in B-hCXCR5 MC38 cells. Human CXCR5 is highly expressed on the surface of B-hCXCR5 MC38 cells.
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Application
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B-hCXCR5 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 CXCR5 coding sequence was inserted to replace part of murine exons 1~2. The insertion disrupts the endogenous murine Cxcr5 gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
CXCR5 expression analysis in B-hCXCR5 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCXCR5 MC38 cultures were stained with an anti-human CXCR5 antibody. Human CXCR5 was detected on the surface of B-hCXCR5 MC38 cells but not wild-type MC38 cells. The 2-A03 clone of B-hCXCR5 MC38 cells was used for in vivo experiments.
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Tumor Growth Curve & Body Weight Changes
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Tumor Growth Curve & Body Weight Changes in C57BL/6 Mice
Subcutaneous homograft tumor growth of B-hCXCR5 MC38 cells. B-hCXCR5 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 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-hCXCR5 MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
