Doege Annika, Steens Rebecca, Dünker Nicole, Busch Maike Anna
Center for Translational Neuro- and Behavioral Sciences (C-TNBS), Institute of Anatomy II, Department of Neuroanatomy, Medical Faculty, University of Duisburg-Essen, 45147 Essen, Germany.
Methods Protoc. 2022 Mar 2;5(2):21. doi: 10.3390/mps5020021.
Retinoblastoma (RB) is a primary intraocular malignancy in childhood. Relapses may develop and cause secondary cancers during later development. This study was set up to identify optimal cell culture conditions for RB cell growth and to optimize tumor growth in an model. RB cell lines (Y79 and WERI-Rb1) were cultivated under three different conditions and apoptosis, proliferation and cell growth, as well as expression profiles of two epithelial-mesenchymal transition (EMT) markers, were analyzed. EMT gene expression profiles were not generally changed, whereas apoptosis levels, tumor cell proliferation, and growth were significantly influenced by different cell culture conditions. In order to optimize the time-limited chick chorioallantoic membrane (CAM) assay, we investigated two different time points of tumor cell inoculation (embryonic development day EDD8 and EDD10) as well as three different cell concentrations. We showed that inoculation at EDD8 led to decreased tumor formation and chicken viability, whereas different cell concentrations did not change size and weight of developing tumors. Our findings demonstrate that medium conditions as well as the starting point for CAM inoculation significantly influence the experimental outcome of investigations using RB cell lines.
视网膜母细胞瘤(RB)是儿童期原发性眼内恶性肿瘤。复发可能会出现,并在后期发展过程中引发继发性癌症。本研究旨在确定视网膜母细胞瘤细胞生长的最佳细胞培养条件,并在一个模型中优化肿瘤生长。视网膜母细胞瘤细胞系(Y79和WERI-Rb1)在三种不同条件下培养,并分析了细胞凋亡、增殖和生长情况,以及两种上皮-间质转化(EMT)标志物的表达谱。EMT基因表达谱一般没有变化,而细胞凋亡水平、肿瘤细胞增殖和生长受到不同细胞培养条件的显著影响。为了优化限时鸡胚绒毛尿囊膜(CAM)试验,我们研究了肿瘤细胞接种的两个不同时间点(胚胎发育第8天和第10天)以及三种不同细胞浓度。我们发现,在胚胎发育第8天接种会导致肿瘤形成减少和鸡的存活率降低,而不同细胞浓度并未改变发育中肿瘤的大小和重量。我们的研究结果表明,培养基条件以及CAM接种的起始点会显著影响使用视网膜母细胞瘤细胞系进行研究的实验结果。