目的 建立视网膜母细胞瘤(retinoblastoma,RB)模型,为研究RB新疗法提供实验动物模型。方法 选取出生第1天的CD1小鼠40只,随机分为A、B、C、D 4组,每组10只。设每组小鼠的右眼为实验眼,左眼为对照眼,将RB-Y79细胞以4 μl/只注射至右眼玻璃体腔内,A、B、C、D组注射的RB-Y79浓度分别是2.0×107/ml、3.0×107/ml、3.5×107/ml、4.0×107/ml。4组小鼠左眼均注射等量重悬细胞的溶剂;观察小鼠生长状态及移植瘤生长情况,并与对照眼比较,选择出最佳建模细胞浓度;取出的组织常规石蜡切片HE染色,观察移植瘤组织细胞形态学特征。结果 注射RB-Y79细胞后,实验小鼠右眼可形成肿瘤,而对照眼左眼无肿瘤形成;注射的RB-Y79细胞浓度不同,成瘤率和存活率不同,第10天A、B、C、D组的成瘤率分别为20%、80%、100%和100%,存活率分别为100%、80%、80%和50%。C组3.5×107/ml为最佳建模浓度;移植瘤组织病理检查:右眼眼球结构被破坏。瘤细胞大小不等,形态为圆形、椭圆形、多边形或不规则形;细胞质少;胞核大,圆形,卵圆形或不规则形,染色深,有1~2个以上核样结构。结论 新生小鼠玻璃体腔内注射RB-Y79细胞可成功建立RB小鼠模型。
Abstract
Objective To establish a retinoblastoma model and provide an animal model for the new treatment of RB. Methods Forty CD1 postnatal day 1 mice were randomly divided into 4 groups ( A,B, C, D), 10 in each. We set the right eye of each mouse as experiment group and the left as control. Different density of RB-Y9 cells(A 2.0×107/ml,B 3.0×107/ml,C 3.5×107/ml,D 4.0×107/ml, 4 μl/one) were injected into the vitreous cavity of right eyes, and the left eyes were injected with the same volume of cell culture medium. We closely monitored the tumor growth after injection and found the most appropriate density of the cells for the model construction. We used HE staining to analyze the tumor formation and histology and convinced that the tumor was RB. Results After the cell injection, we found no tumor formation in control group and most mice in the experiment group had tumors. Different density of injeced cells resulted in different rates of survival and tumor formation: the rate of tumor formation in the above 4 groups were 20%,80%,100% and 100% and the survival rates were100%,80%,80% and 50%. We regarded the C group (the density of the cells was 3.5×107/ml) as the best model for the experiment. HE staining of the tumor tissues implied that the structure of right eyes were severely destroyed and the cells had different size and were irregularly shaped: some were circular, oval and others were polygonal or irregular. The tumor cells had less cytoplasm and bigger nuclei when compared with normal controls. Cells with more than 1-2 nuclei can be frequently observed. Conclusions By injecting RB-Y79 cells in the vitreous cavity of neonatal mice can successfully develop the RB mouse model.
关键词
CD1新生小鼠 /
视网膜母细胞瘤 /
移植瘤模型
Key words
CD1 postnatal day 1mice /
retinoblastoma /
transplanted tumor model
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