Basic Information
Description
The mouse TF gene was replaced by the human TF coding sequence in B-hTF MC38 cells. Human TF is highly expressed on the surface of B-hTF MC38 cells.
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Application
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B-hTF 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 TF coding sequence was inserted to replace part of murine exon 3 and all of exons 4~5. The insertion disrupts the endogenous murine TF gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
TF expression analysis in B-hTF MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hTF MC38 cultures were stained with species-specific anti-TF antibodies. Human TF was detected on the surface of B-hTF MC38 cells but not wild-type MC38 cells. The 2-D03 clone of B-hTF MC38 cells was used for in vivo experiments.
Post-inoculation
B-hTF MC38 cells were subcutaneously implanted into C57BL/6 mice (n=5). Tumor cells were harvested on day 31 post inoculation and assessed for TF expression by flow cytometry using species-specific antibodies. As shown, human TF was highly expressed on the surface of humanized B-hTF MC38 tumor cells, indicating B-hTF MC38 cells can be used for in vivo efficacy studies of novel TF therapeutics.
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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-hTF MC38 cells. B-hTF 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-hTF MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
