Targeting SOX17 in Human Embryonic Stem Cells Creates Unique Strategies for Isolating and Analyzing Developing Endoderm

Wang, Pei; Rodriguez, Ryan T.; Wang, Jing; Ghodasara, Amar; Kim, Seung K.

doi:10.1016/j.stem.2011.01.017

Cited by 132 publications

(103 citation statements)

References 42 publications

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“…Because activin A-induced differentiation does not generate homogenous cell population (Supplementary information, Figure S1B), we used CD184 (also named CXCR4), a well-known definitive endoderm marker [4,[19][20][21], to sort the cell population and confirmed successful endoderm differentiation by RT-qPCR analysis (Supplementary information, Figure S1C). In addition to CD184, another report [20] used both CD184 and CD117 (also named c-KIT) to mark the definitive endoderm cell population.…”

Section: Resultsmentioning

confidence: 99%

Histone H3K27me3 demethylases KDM6A and KDM6B modulate definitive endoderm differentiation from human ESCs by regulating WNT signaling pathway

2012

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Definitive endoderm differentiation is crucial for generating respiratory and gastrointestinal organs including pancreas and liver. However, whether epigenetic regulation contributes to this process is unknown. Here, we show that the H3K27me3 demethylases KDM6A and KDM6B play an important role in endoderm differentiation from human ESCs. Knockdown of KDM6A or KDM6B impairs endoderm differentiation, which can be rescued by sequential treatment with WNT agonist and antagonist. KDM6A and KDM6B contribute to the activation of WNT3 and DKK1 at different differentiation stages when WNT3 and DKK1 are required for mesendoderm and definitive endoderm differentiation, respectively. Our study not only uncovers an important role of the H3K27me3 demethylases in definitive endoderm differentiation, but also reveals that they achieve this through modulating the WNT signaling pathway.

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Section: Resultsmentioning

confidence: 99%

Histone H3K27me3 demethylases KDM6A and KDM6B modulate definitive endoderm differentiation from human ESCs by regulating WNT signaling pathway

2012

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“…Wang et al established a Sox17/GFP knock-in human ES cell line, and carried out gene expression analysis of Sox17/GFP+ cells that were differentiated based on the procedure established by the D'Amour group. The results of their gene expression analysis, in vitro differentiation, and transplantationbased assays showed that CD49e+CD141+CD238+ cells are primitive gut tube endoderm cells [26]. Human ES cell lines were established, with a Sox17/GFP or Pdx1/GFP transgene introduced via BAC vectors.…”

Section: Gene Profiling Of Human Es Cell-derived Cellsmentioning

confidence: 99%

“…G protein-coupled receptor 50 (GPR50) and tumorassociated calcium signal transducer 2 (TROP2) were identified as cell surface proteins that were highly enriched in pancreatic progenitor cells [27]. The identification of cell surface marker genes enabled the isolation of DE [11,28], primitive gut tube endoderm [26], and pancreatic progenitor cells [27], without genetic manipulation of ES cells. This method represents a powerful tool for future characterization of similar cell populations.…”

Section: Gene Profiling Of Human Es Cell-derived Cellsmentioning

confidence: 99%

Profiling of Embryonic Stem Cell Differentiation

2014

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■ AbstractEmbryonic stem (ES) cells have been shown to recapitulate normal developmental stages. They are therefore a highly useful tool in the study of developmental biology. Profiling of ES cell-derived cells has yielded important information about the characteristics of differentiated cells, and allowed the identification of novel marker genes and pathways of differentiation. In this review, we focus on recent results from profiling studies of mouse embryos, human islets, and human ES cell-derived differentiated cells from several research groups. Global gene expression data from mouse embryos have been used to identify novel genes or pathways involved in the developmental process, and to search for transcription factors that regulate direct reprogramming. We introduce gene expression databases of human pancreas cells (Beta Cell Gene Atlas, EuroDia database), and summarize profiling studies of islet-or human ES cell-derived pancreatic cells, with a focus on gene expression, microRNAs, epigenetics, and protein expression. Then, we describe our gene expression profile analyses and our search for novel endoderm, or pancreatic, progenitor marker genes. We differentiated mouse ES cells into mesendoderm, definitive endoderm (DE), mesoderm, ectoderm, and Pdx1-expressing pancreatic lineages, and performed DNA microarray analyses. Genes specifically expressed in DE, and/or in Pdx1-expressing cells, were extracted and their expression patterns in normal embryonic development were studied by in situ hybridization. Out of 54 genes examined, 27 were expressed in the DE of E8.5 mouse embryos, and 15 genes were expressed in distinct domains in the pancreatic buds of E14.5 mouse embryos. Akr1c19, Aebp2, Pbxip1, and Creb3l1 were all novel, and none has been described as being expressed, either in the DE, or in the pancreas. By introducing the profiling results of ES cell-derived cells, the benefits of using ES cells to study early embryonic development will be discussed.

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“…Expression data were obtained from the Gene Expression Omnibus (GEO) database. 42 The used data sets 19,[43][44][45][46][47][48][49][50][51][52][53][54][55] are summarized in Table 1. Primary data analysis was performed with Expression Console (Affymetrix) applying the Microarray Suite 5 algorithm.…”

Section: Cells and Cell Culture Pbmc DC Eft Cell Lines A673mentioning

confidence: 99%