Establishment of orthotopic model of SHG-44 in enhanced green fluorescent protein nude mice

Objective To establish the orthotopic model of SHG-44 in the enhanced green fluorescent protein (EGFP) nude mice.Methods The C57BL/6J-EGFP transgenic mice were crossed with nude mice,then the hairless male nude mouse expressing EGFP was crossed with the hairy female pubescence mouse to obtain the EGFP nude mouse.The expression status of EGFP in the EGFP nude mice was evaluated under the fluorescence microscopy and by using polymerase chain reaction (PCR).The human glioma cell line SHG-44 cells ( 15 μL cell suspensions,including 1 × 106 cells) were stereotaxically implanted into the brain of the EGFP nude mouse to produce orthotopic models.Results Each mouse could give birth to 7-8 litter size on average,and there were 3-4 naked young homozygous mice,the ratio close to 1∶ 1.Through the small animal in vivo imaging system 2 days after birth,the expression of EGFP gene could be identified.Green fluorescence was seen in the frozen sections of organs and tissues under the fluorescent microscopy.PCR results showed that the brain tissue,subcutaneous muscle,tail and toes all expressed the EGFP gene.The tumor formation rate of the orthotopic model was about 100％.The cells of orthotopic model still had the basic characteristics of human glioma cells.Conclusion The EGFP nude mouse can be used for human tumors transplantation experiments. Key words: Green fluoresent protein; Nude mice; Transgenic animal