Establishment Of EGFP Immunodeficient Congenic Inbred Mice Strain And Preliminary Application In Glioma Xenograft Model

Part Ⅰ: Establishment and verification of new congenic inbred mouse strain namedFoxn1nu.B6-CAG-EGFP/SU and NOD/SCID /EGFPObject To establish a new inbred mouse strain carrying and expressing EGFP and Foxn1 nu gene for cancer research, including human glioma as well.Methods According to criterion of GB14923-2010, the male Foxn1 nu nude mice backcross female C57BL/6-Tg(CAG-EGFP) transgenic mice for 10 times. Then identify the phenotype using the methods and equipment as below: fluorescent flashlight and matching glasses; multifunction vivo imager; fluorescence microscopy.Results The inbred strain named Foxn1 nu.B6-CAG-EGFP/SU(Soochow University). All the 14 biochemical loci are homozygous, yet in compare with the BALB/c strain, one biochemical loci “Pep3” is still different(“b” type instead of “a” type). Peripheral blood lymphocyte count shows the lymphocytes occupy 15% of nucleated cells; T lymphocytes occupy 0.3%, meet the requirement of inbred strain of EGFP nude mice.Conclusions Established a new inbred strain-Foxn1 nu.B6-CAG-EGFP/SU, which both express EGFP stably, and own immunodeficiency with lack of T lymphocytes. The phenotype “b” of biochemical loci “Pep3” is the unique characteristic that distinguishes SU to Foxn1 nu.Part Ⅱ: Analysis of fluorescence expression in different brain regions of EGFP inbred mice and its application in brain tumor transplantation modelObject: To analyze the expression of EGFP in different brain regions of inbred line for brain tumor orthotopic transplantation research.Methods: The expression of EGFP in brain tissue was detected by immunofluorescence assay. Establish an orthotopic transplantation model of U87- RFP in BALB/c inbred strain. A brain metastasis tumor tissue of clinical case was inoculated into NOD/SCID/EGFP mice, and the orthotopic transplantation tumor model was established.Results: The EGFP espress stably and intensively in the brain of the novel inbred strain. And the success rate of tumor transplantation in inbred strain was 100%. Besides, the cells of NOD/SCID/EGFP mice inoculated clinical samples were found to be well preserved in this model.Conclusion: The new inbred strain express EGFP stably and intensively, which can be used as a foundation for the glioma and tumor microenvironment research. NOD/SCID/EGFP mice could be a better host for clinical tumor specimens.