Basic Information
Description
Immunodeficient B-NDG mice were generated by deleting the IL2rg gene from NOD-scid mice. This mouse model has the highest degree of immunodeficiency and is most suitable for engraftment and growth of human tumor cells or tissues.
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Targeting strategy
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Gene targeting strategy for B-NDG hTHPO mice.
The full sequences of mouse Thpo gene except the UTRs were replaced with the coding sequences (CDS) of human THPO gene.
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mRNA expression analysis
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Strain specific analysis of THPO gene expression in C57BL/6 mice, B-NDG mice and B-NDG hTHPO mice by RT-PCR. Mouse Thpo mRNA was detectable in liver and kidney of C57BL/6 mice (+/+) and B-NDG mice (+/+). Human THPO mRNA was detectable only in homozygous B-NDG hTHPO mice (H/H) but not in wild-type C57BL/6 mice and B-NDG mice.
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Human immune system engraftment
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Human CD34+ HSCs (5E5) were intravenous engrafted into wild-type B-NDG mice and homozygous B-NDG hTHPO mice (female, 6 week-old, n=20). B-NDG mice were treated with 2Gy-irradiation. B-NDG hTHPO mice were not irradiated. Flow cytometric analysis of peripheral blood lymphocyte from the two mice after engraftment with human CD34+ HSCs were shown in the figure above. Results showed that the survival rate of B-NDG hTHPO mice was higher than that of B-NDG mice. The percentages of human CD45+ leukocytes, T cells, B cells, NK cells, myeloid cells, monocytes, macrophages and neutrophils in B-NDG hTHPO mice were similar to those in B-NDG mice, demonstrating that the humanized mouse model engrafted with human CD34+ HSCs was successfully constructed. Values are expressed as mean ± SEM. HSCs: hematopoietic stem cells.
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Summary
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- mRNA expression analysis:
Mouse Thpo mRNA was detectable in liver and kidney of C57BL/6 mice and B-NDG mice. Human THPO mRNA was detectable in homozygous B-NDG hTHPO mice.
- Human immune system engraftment ：
The survival rate of B-NDG hTHPO mice was higher than that of B-NDG mice. The percentages of human CD45+ leukocytes, T cells, B cells, NK cells, myeloid cells, monocytes, macrophages and neutrophils in B-NDG hTHPO mice were similar to those in B-NDG mice, demonstrating that the humanized mouse model engrafted with human CD34+ HSCs was successfully constructed.
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References
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1.Kuter, D. J. The biology of thrombopoietin and thrombopoietin receptor agonists. International journal of hematology 98, 10-23, doi:10.1007/s12185-013-1382-0 (2013).
2.Rongvaux A, et al., Human thrombopoietin knockin mice efficiently support human hematopoiesis in vivo. Proc Natl Acad Sci U S A. 2011 Feb 8;108(6):2378-83
