Basic Information
Description
The mouse Fgl1 gene was replaced by the human FGL1 coding sequence in B-hFGL1 MC38 cells. Human FGL1 is highly expressed and secreted by B-hFGL1 MC38 cells.
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Application
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B-hFGL1 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 FGL1 coding sequence was inserted to replace part of murine exon 4 and all of exons 5-7. The insertion disrupts the endogenous murine Fgl1 gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
FGL1 expression analysis in B-hFGL1 MC38 cells by ELISA. Supernatant was collected from wild-type MC38 and B-hFGL1 MC38 cultures and analyzed using a species-specific FGL1 ELISA kit. Human FGL1 was detectable in the supernatant of B-hFGL1 MC38 cells but not in that of wild-type MC38 cells. The 3-A07 clone of B-hFGL1 MC38 cells was used for in vivo experiments.
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Tumor Growth Curve
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Tumor Growth Curve & Body Weight Changes in C57BL/6N Mice
Subcutaneous homograft tumor growth of B-hFGL1 MC38 cells. B-hFGL1 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N mice (female, 8-week-old, n=6). (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-hFGL1 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
