Array comparative genomic hybridization of spheres derived from TH-MYCN (hemizygote) mice [Agilent-027441]

It has been already reported that there are undifferentiated/proliferating neuroblasts in the postnatal sympathetic ganglia in TH-MYCN mice, a neuroblastoma model. We established suitable spheroid culture condition that selectively isolates undifferentiated neuroblasts from superior mesenteric ganligon (SMG) of TH-MYCN mice. In order to investigate the chromosomal alterations (gains or losses) of spheres derived from TH-MYCN mice, we carried out array comparative genomic hybridization. We investigated whether chromosomal alterations occured during early neuroblastoma tumorigenesis in TH-MYCN mice. Primary spheres were cultured from SMG of E13.5 and 3-week-old TH-MYCN mice, and from tumor tissues of TH-MYCN mice. Eight biological replicates of primary spheres from tumor tissues of TH-MCYN mice were subjected to the array CGH. Passageable spheres were obtained by subculturing primary spheres derived from E13.5 and 3-week-old TH-MYCN SMG. Six biological replicates of E13.5 TH-MYCN spheres and eight biological replicates of 3-week-old TH-MYCN spheres were subjected to array CGH.

已有研究证实，作为神经母细胞瘤模型的TH-MYCN小鼠的产后交感神经节内存在未分化/增殖性成神经细胞。本研究建立了可从TH-MYCN小鼠肠系膜上神经节（SMG）中选择性分离未分化成神经细胞的适配球体培养体系。为探究TH-MYCN小鼠来源球体的染色体畸变（拷贝数增加或缺失）特征，我们开展了阵列比较基因组杂交（array CGH）实验，以明确染色体畸变是否发生于TH-MYCN小鼠神经母细胞瘤的早期肿瘤发生过程中。我们分别从E13.5胎龄及3周龄TH-MYCN小鼠的肠系膜上神经节，以及TH-MYCN小鼠的肿瘤组织中分离培养原代球体。针对TH-MYCN小鼠肿瘤组织来源的原代球体，我们设置了8个生物学重复样本用于array CGH检测。通过对E13.5胎龄与3周龄TH-MYCN小鼠肠系膜上神经节来源的原代球体进行传代培养，成功获得可稳定传代的球体。针对E13.5胎龄TH-MYCN小鼠球体及3周龄TH-MYCN小鼠球体，我们分别设置6个和8个生物学重复样本用于array CGH检测。