Basic Information
Description
The mouse Mpl gene was replaced by human c-MPL coding sequence in B-hc-MPL MC38 cells. Human c-MPL is highly expressed on the surface of B-hc-MPL MC38 cells.
-
Targeting strategy
-
Gene targeting strategy for B-hc-MPL MC38 cells. The exogenous promoter and human c-MPL coding sequence was inserted to replace part of murine exon 3 and all of exon 4. The insertion disrupts the endogenous murine Mpl gene, resulting in a non-functional transcript.
-
Protein expression analysis
-
c-MPL expression analysis in B-hc-MPL MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hc-MPL MC38 cultures were stained with species-specific anti-c-MPL antibody. Human c-MPL was detected on the surface of B-hc-MPL MC38 cells but not wild-type MC38 cells. The 1-H01 clone of B-hc-MPL MC38 cells was used for in vivo experiments.
-
Tumor growth curve & Body weight changes
-
Subcutaneous homograft tumor growth of B-hc-MPL MC38 cells. B-hc-MPL MC38 cells (5×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 11-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hc-MPL MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
-
Protein expression analysis of tumor cells
-
B-hc-MPL MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6), and on 35 days post inoculation, tumor cells were harvested and assessed for human c-MPL expression by flow cytometry. As shown, human c-MPL was highly expressed on the surface of tumor cells. Therefore, B-hc-MPL MC38 cells can be used for in vivo efficacy studies of novel c-MPL therapeutics.
