Expression data of human eGFP-SOX17+ endoderm cells. Homo sapiens

Using homologous recombination in human ESC, we inserted an enhanced green fluorescent protein (eGFP) transgene into a locus encoding a postulated marker of human endoderm, SOX17 in H9 human embryonic stem cells. This allowed purification of SOX17+ hESC endodermal progeny by fluorescence activated cell sorting (FACS) to generate microarray gene expression profile. Overall design: Using Wnt3 and Activin to differentiate hSOX17-2 to stage 1 cells, and subsequently FGF10 and cyclopamine to stage 2 cells, we isolated eGFP+ cells by FACS at each stage, performed microarray analysis.

本研究借助人类胚胎干细胞（human embryonic stem cells, hESC）的同源重组技术，将增强型绿色荧光蛋白（enhanced green fluorescent protein, eGFP）转基因插入至H9人类胚胎干细胞中编码人类内胚层推定标记物SOX17的基因座。该策略可通过荧光激活细胞分选术（fluorescence activated cell sorting, FACS）纯化SOX17阳性的人类胚胎干细胞内胚层子代细胞，以制备基因表达微阵列谱。实验整体设计：采用Wnt3与激活素（Activin）诱导hSOX17-2细胞分化为第1阶段细胞，后续辅以成纤维细胞生长因子10（FGF10）与环巴胺处理得到第2阶段细胞；于每个阶段均通过FACS分离eGFP阳性细胞，并开展微阵列基因表达分析。