Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD93 mice. The exon 1 of mouse Cd93 gene that encode the extracellular domain was replaced by human CD93 exon 1 in B-hCD93 mice.
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Protein expression analysis
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Strain specific CD93 expression analysis in heterozygous B-hCD93 mice by flow cytometry. Bone marrow were collected from wild type and heterozygous B-hCD93 mice (H/+) , and analyzed by flow cytometry with species-specific CD93 antibody. Mouse CD93 was detectable in wild type mice and heterozygous B-hCD93 mice. Human CD93 was exclusively detectable in heterozygous B-hCD93 mice but not in wild type mice.
Strain specific CD93 expression analysis in homozygous B-hCD93 mice by flow cytometry. Bone marrow were collected from C57BL/6 wild type and homozygous B-hCD93 mice (H/H) , and analyzed by flow cytometry with species-specific CD93 antibody. Mouse CD93 was detectable in wild type mice. Human CD93 was exclusively detectable in homozygous B-hCD93 mice but not in wild type mice.
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Immune Cell Profile
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Analysis of leukocytes cell subpopulation in spleen
Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hCD93 mice (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hCD93 mice were similar to those in the C57BL/6 mice, demonstrating that CD93 humanized does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.
Analysis of T cell subpopulation in spleen
Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hCD93 mice (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hCD93 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD93 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.
Analysis of leukocytes cell subpopulation in lymph node
Analysis of lymph node leukocyte subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hCD93 mice (n=3, 7-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, and NK cells in homozygous B-hCD93 mice were similar to those in the C57BL/6 mice, demonstrating that CD93 humanized does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.
Analysis of T cell subpopulation in lymph node
Analysis of lymph node T cell subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hCD93 mice (n=3, 7-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hCD93 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD93 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.
Analysis of leukocytes cell subpopulation in blood
Analysis of blood leukocyte subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hCD93 mice (n=3, 7-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hCD93 mice were similar to those in the C57BL/6 mice, demonstrating that CD93 humanized does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.
Analysis of T cell subpopulation in blood
Analysis of blood T cell subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hCD93 mice (n=3, 7-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hCD93 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD93 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.
