视网膜母细胞瘤细胞系裸小鼠玻璃体腔的异种移植

目的 建立裸小鼠玻璃体腔异种视网膜母细胞瘤(retinoblastoma,RB)移植瘤模型，用于RB的体内基因治疗试验。 方法 将来源于人的RB细胞系HXO-Rb₄₄制成细胞悬液注射于10只裸小鼠20只眼的玻璃体腔。随机将裸小鼠分为2组，第1组5只鼠10只眼，每只眼注5μl；第2组5只鼠10只眼，每只眼注10μl。用肉眼和直接检眼镜对裸小鼠RB模型作活体观察和半定量分级；用光镜、电镜对移植瘤作组织病理形态观察；用卵白素-生物素-过氧化物酶复合法(avidin-biotin-complex method,ABC法)进行神经元特异性烯醇酶(neuron specific enolase,NSE)、神经胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的免疫组织化学染色检查和流式细胞仪测定其DNA指数(DNA index,DI)及S期细胞百分比(s-phase fraction,SPF)。 结果 在裸小鼠眼玻璃体腔内均成功地形成了移植性RB肿瘤。移植瘤对NSE呈阳性反应，对GFAP呈阴性反应。DI值大于1.1,SPF检测表明移植瘤细胞有增殖活性。结论 裸小鼠玻璃体腔内RB异种移植瘤形态学和免疫组织化学特征均与人类RB细胞相似，是进一步进行RB治疗研究的一个较好动物模型。(中华眼底病杂志,1998,14:144-148)

Heterotransplantation of retinoblastoma cell line into the vitreous cavity of nude mice

PurposeTo establish orthotopic heterotransplanted model of retinoblastoma(RB)into the vitreous cavity of nude mice for experimental studies of gene therapies in vivo.Methods To prepare the concentrated cell suspensions of human RB cell line HXO-Rb₄₄ and inject them into the vitreous cavities of 10 nude mice(20 eyes).The experimental mice were randomly divided into 2 groups:the first group,5 mice (10 eyes) were injected with 5 μl of the cell suspensions,and the second group,5 mice (10 eyes)with 10 μl．The transplanted RB was observed by inspection and ophthalmoscopy,and graded semi-quantitatively in vivo.Light and electrone microscopes were used for histopathological examination,ABC immunohistochemical methods for NSE and GFAP,and flow cytometric investigations for DNA index(DI) and s-phase fraction (SPF). ResultsAll of the models of RB were successfully established by injecting the cell suspensions of human RB cell line into the mice vitreous cavities.The immunohistochemical reaction of the transplanted tumor cells to NSE was positive,and negative to GFAP．The DNA index was found to be more than 1.1,and the determination of SPF indicated the presence of proliferative ability of the transplanted RB cells. ConclusionThe above findings reveal that the transplanted tumor cells were similar to those of the parent cells,hence the orthotopic heterotransplantation of RB via nude mice vitreous cavity injection is a superior method in establishing the animal model for the further experimental studies of gene therapies in vivo.(Chin J Ocul Fundus Dis,1998,14:144-148)