CAJ | 학술논문

目的建立SHG-44绿色荧光裸鼠原位模型.方法将C57BL/6J-增强绿色荧光蛋白(ECFP)转基因小鼠与裸小鼠进行交配,在继代时严格挑选都表达EGFP基因的无毛雄鼠(♂)与有毛母鼠(♀)交配,通过肉眼、荧光显微镜和分子生物学等方法观察EGFP基因在裸小鼠全身各组织中的表达；将胶质瘤细胞株SHG-44(15μl,含1×106个细胞)注射于绿色荧光裸鼠的脑内制成原位模型.结果每只种鼠平均产仔7～8只,其中纯合裸仔鼠有3～4只,比例接近1∶1；每代仔鼠通过小动物活体成像系统出生后2d就可鉴别其是否表达EGFP基因；器官及组织冰冻切片荧光显微镜下观察均见有绿色荧光；聚合酶链反应(PCR)结果表明脑组织、皮下肌肉、鼠尾和脚趾全部表达EGFP基因；原位模型的成瘤率达100％,瘤细胞依然具有人脑胶质瘤的基本特征.结论表达EGFP基因的裸小鼠可用于肿瘤移植实验.
목적 건립SHG-44록색형광라서원위모형.방법 장C57BL/6J-증강록색형광단백(ECFP)전기인소서여라소서진행교배,재계대시엄격도선도표체EGFP기인적무모웅서(♂)여유모모서(♀)교배,통과육안、형광현미경화분자생물학등방법관찰EGFP기인재라소서전신각조직중적표체；장효질류세포주SHG-44(15μl,함1×106개세포)주사우록색형광라서적뇌내제성원위모형.결과 매지충서평균산자7～8지,기중순합라자서유3～4지,비례접근1∶1；매대자서통과소동물활체성상계통출생후2d취가감별기시부표체EGFP기인；기관급조직빙동절편형광현미경하관찰균견유록색형광；취합매련반응(PCR)결과표명뇌조직、피하기육、서미화각지전부표체EGFP기인；원위모형적성류솔체100％,류세포의연구유인뇌효질류적기본특정.결론 표체EGFP기인적라소서가용우종류이식실험.
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.