Basic Information
Description
The mouse Alb gene was replaced by human ALB coding sequence in B-hALB MC38 cells. Human ALB can be secreted by B-hALB MC38 cells.
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Targeting strategy
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Gene targeting strategy for B-hALB MC38 cells. The exogenous promoter and human ALB coding sequence was inserted to replace part of murine exon 1~2. The insertion disrupts the endogenous murine Alb gene, resulting in a non-functional transcript.
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Protein expression analysis
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ALB expression analysis in B-hALB MC38 cells by ELISA. (A) Mouse ALB was detected in the supernatant of B-Tg(mAlb) MC38 cells but not wild-type MC38 cells. It has been verified that MC38 cells do not express the mouse ALB. (B) Human ALB was detected in the supernatant of B-hALB MC38 cells but not wild-type MC38 cells. The 3-B01 clone of B-hALB MC38 cells was used for in vivo experiments.
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Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-hALB MC38 cells. B-hALB MC38 cells (1×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N mice (female, 6-week-old, n=5). 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-hALB MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
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Protein expression analysis of tumor cells
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B-hALB MC38 cells were subcutaneously transplanted into C57BL/6N mice (n=5), and on 35 days post inoculation, tumor cells were harvested and assessed for mouse and human ALB expression by ELISA. As shown, human ALB was highly expressed tumor homogenate. Therefore, B-hALB MC38 cells can be used for in vivo efficacy studies of novel ALB therapeutics.
