Project description:Our RNA-seq results show the changes in gene expression during differentiation and maturation of iPS-derived enterocytes, as well as that treatment during the last stage with Vitamin D3 (VD3) enhanced enterocyte-related gene expression. Caco-2 and RNA from human adult small intestine were also sequenced as comparison.
Project description:Our RNA-seq results showed that valproic acid (VPA) works synergistically with activated Vitamin D3 (VD3) to enhance the expression of enterocyte-function-related genes in iPS-derived enterocyte-like cells.
Project description:We report the application of MNase-seq (Kent, Adams et al. 2011) to construct genome-wide nucleosome maps from human cells. To date, genome-wide changes in chromatin structure that occur during development in human cells have not been investigated widely. We have constructed and compared genome-wide chromatin maps from undifferentiated human iPS cells and and iPS cells differentiated to neural progenitor cells (NPC).
Project description:Here, we report the generation of human induced Pluripotent Stem (iPS) cell reporter line in which a venus fluorescent protein have been introduced into the PAX7 locus. We use microarrays to compare the transcriptome of PAX7-venus+ cells after 3 weeks of myogenic differentiation to that of undifferentiated iPS Satellite cells (SC) are muscle stem cells which can regenerate adult muscles upon injury. Most SC originate from PAX7-positive myogenic precursors set aside during development. While myogenesis has been studied in mouse and chicken embryos, little is known about human muscle development. Here, we use microarrays to compare the transcriptome of myogenic cells differentiated in vitro from human and mouse ES and iPS cells reporter cell lines. We generated fluorescent reporter lines in which a fluorescent protein was introduced into the loci coding for Pax7 (mouse ES and human iPS) or MYOG (human iPS). Mouse ES and human iPS cells were differentiated to the myogenic lineage in vitro for three weeks according to the protocols described in Chal et al, Nature Biotech 2015 and to Chal, Al Tanoury et al, Nature Protocols, 2016. Fluorescent Pax7 or MYOG-positive cells were FACS-sorted after 3-weeks of differentiation in vitro and we used Affymetrix microarrays to analyze the transcriptome of the purified mouse and human cell populations and compare it to the transcriptome of undifferentiated mouse ES or human iPS cells.
Project description:Here, we report the generation of human induced Pluripotent Stem (iPS) cell reporter line in which a venus fluorescent protein have been introduced into the MYOGENIN (MYOG) locus. We use microarrays to compare the transcriptome of MYOG-venus+ cells after 3 weeks of myogenic differentiation to that of undifferentiated iPS Satellite cells (SC) are muscle stem cells which can regenerate adult muscles upon injury. Most SC originate from PAX7-positive myogenic precursors set aside during development. While myogenesis has been studied in mouse and chicken embryos, little is known about human muscle development. Here, we use microarrays to compare the transcriptome of myogenic cells differentiated in vitro from human and mouse ES and iPS cells reporter cell lines. We generated fluorescent reporter lines in which a fluorescent protein was introduced into the loci coding for Pax7 (mouse ES and human iPS) or MYOG (human iPS). Mouse ES and human iPS cells were differentiated to the myogenic lineage in vitro for three weeks according to the protocols described in Chal et al, Nature Biotech 2015 and to Chal, Al Tanoury et al, Nature Protocols, 2016. Fluorescent Pax7 or MYOG-positive cells were FACS-sorted after 3-weeks of differentiation in vitro and we used Affymetrix microarrays to analyze the transcriptome of the purified mouse and human cell populations and compare it to the transcriptome of undifferentiated mouse ES or human iPS cells.
Project description:This experiment was designed to show the similarity among normal human epidermal melanocytes, melanocytes derived from human 3F-induced pluripotent stem (iPS) cells, and human 3F-iPS cells. Human 3F-iPS cells without c-myc were established and then differentiated into melanocytes. The differentiated cells were collected and the gene expression profile was compared to normal human epidermal melanocytes (NHEMs) and orginal 3F-iPS cells.
Project description:We used microarrays to detail the differentail gene expression between intestinal Lgr5(hi) stem cells and differentiated cells Assay the differential gene expression using total RNA from flow cytometry sorted Lgr5(hi) cells and EDTA isolated enterocytes from Atoh1 conditional knockout
Project description:Hepatocyte-like cells differentiated from human iPS cells are expected to be utilized in pharmaceutical research and regenerative medicine. Recently, various culture methods for human iPS cell maintenance have been developed. However, it is not well known whether human iPS cell maintenance method affects hepatic differentiation potency. In this study, we cultured human iPS cells using four maintenance methods: ReproStem medium with feeder cells (mouse embryonic fibroblasts), AK02N medium with iMatrix-511 (E8 fragments of laminin511), Essential 8 medium with Vitronectin N (N-terminal domain of vitronectin), TeSR-E8 medium with Vitronectin XF (xeno-free vitronectin). Then, these human iPS cells were differentiated into the hepatocyte-like cells. Interestingly, the gene expression levels of definitive endoderm markers in the definitive endoderm cells generated from human iPS cells cultured with ReproStem or TeSR-E8 medium were higher than those in other groups. The gene expression level of foregut marker, HHEX, in the definitive endoderm cells generated from human iPS cells cultured with ReproStem medium was higher than that in other groups. Consistently, the expression levels of hepatocyte markers, albumin and urea secretion capacity, and CYP3A4 activity in the hepatocyte-like cells generated from human iPS cells cultured with ReproStem medium were higher than those in other groups. Our data indicated that the most suitable human iPS cell maintenance method for efficient hepatic differentiation was the on-feeder method which uses mouse embryonic fibroblasts, but not feeder-free methods. In conclusion, human iPS cell maintenance method largely affects hepatic differentiation potency.