摘要
目的运用异体移植动物模型及活体成像技术,构建人子宫内膜异位症的可示踪裸鼠模型。方法采集人子宫在位内膜组织,制成悬液。应用慢病毒载体将荧光素酶和红色荧光蛋白基因转入内膜组织并培养48h,将等体积的悬液(分成3组:A组已转染的内膜组织,B组未转染的内膜组织,C组等体积的DMEM/F12培养基)注射到BALB/C雌性裸鼠腹腔内,运用活体成像仪监测异位病灶的发展情况。28d后分离异位病灶,运用苏木素-伊红(HE)染色和免疫组织化学鉴定异位病灶的来源并分析成模情况。结果已成功转染内膜组织的裸鼠在活体成像仪监测下显示,异位病灶的荧光信号在种植后第7天至第14天减小(P<0.01);第14天到第28天增大(P<0.05)。HE染色和免疫组织化学显示异位病灶均来自于人子宫内膜组织。结论人子宫内膜异位症的可示踪裸鼠模型构建成功。
Objective To establish tracing nude mouse model of human endometriosis,which was transfected by lentivirus-RFP-luciferase and detected by in vivo bioluminescence imaging(BLI)system.Methods Human endometrial tissues were collected and made into suspension,which were successfully transfected by lentivirus-RFP-luciferase and cultured for 48 hin vitro.Isometric suspension,which was divided into three groups:the transfected group(group A),the untransfected group(group B)and the DMEM/F12 group(group C),was intraperitoneally injected into female BALB/C nude mice.The development of ectopic lesion was dynamically detected by BLI system.Twenty-eight days later,in order to learn the establishment of model,ectopic lesions were separated and determinated by biotechnology such as HE stain and immunohistochemical analysis.Results The BLI system showed that the ROI of ectopic lesions decreased from the 7 th to the14 th day(P<0.01),and increased from the 14 th to the 28 th day in group A(P<0.05).Both HE stain and immunohistochemical analysis showed that the ectopic lesions were derived from human endometrial tissues.Conclusion The tracing nude mouse model of human endometriosis is successfully established.
引文
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