Basic Information
Description
The mouse Cd36 gene was replaced by the human CD36 coding sequence in B-hCD36 MC38 cells. Human CD36 is highly expressed on the surface of B-hCD36 MC38 cells.
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Application
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B-hCD36 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 CD36 coding sequence was inserted to replace part of mouse Cd36. The insertion disrupts the endogenous murine Cd36 gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
CD36 expression analysis in B-hCD36 MC38 cells by flow cytometry. Single cell suspensions from B-hCD36 MC38 cultures were stained with an anti-human CD36 antibody. Human CD36 was detected on the surface of B-hCD36 MC38 cells but not wild-type MC38 cells. The 10-G12 clone of B-hCD36 MC38 cells was used for in vivo experiments.
Post-inoculation
B-hCD36 MC38 cells were subcutaneously implanted into B-hCD36 mice (n=5). Tumor cells were harvested on day 35 post inoculation and assessed for human CD36 expression by flow cytometry. As shown, human CD36 was highly expressed on the surface of humanized B-hCD36 MC38 tumor cells, indicating B-hCD36 MC38 cells can be used for in vivo efficacy studies of novel CD36 therapeutics. *Note: B-hCD36 MC38 cells can not establish tumors in wild-type C57BL/6 mice.
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Tumor Growth Curve
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Tumor Growth Curve & Body Weight Changes in Humanized Mice
Subcutaneous homograft tumor growth of B-hCD36 MC38 cells. B-hCD36 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into B-hCD36 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-hCD36 MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies. *Note: B-hCD36 MC38 cells can not establish tumors in wild-type C57BL/6 mice.
