A SIX6 reporter for studies of early optic development

Human pluripotent stem cells represent an exciting tool to investigate the mechanisms of human eye development and to build human models of retinal dystrophy. To explore the dynamics of early eye development in a carefully controlled setting and optimize this process, we have developed an eyefield expressed SIX6-GFP reporter that enables the systematic optimization of early eye-field development allowing the identification of conditions that favor early optic development. Pooled day 65 retina cups (n>3/sample) were separated as either GFP- (n=4), GFP+ (n=4) or SIX6+/POU4F2+ (n=4) samples (total of 12 samples run for RNA-seq).

人类多能干细胞是研究人类眼球发育机制、构建视网膜营养不良疾病人体模型的极具潜力的研究工具。为在严格受控的实验环境下探究早期眼球发育的动态变化并优化该过程，我们构建了眼域特异性表达SIX6基因标记绿色荧光蛋白（SIX6-GFP）的报告细胞系，可系统性优化早期眼域发育进程，从而筛选出有利于早期视神经发育的培养条件。将培养至第65天的混合视网膜杯样本（每样本包含＞3个视网膜杯）分为三组：绿色荧光蛋白阴性（GFP-，n=4）、绿色荧光蛋白阳性（GFP+，n=4）以及SIX6+/POU4F2+双阳性样本（n=4），共计12份样本用于转录组测序（RNA-seq）。