Pancreatic islet, pancreas and gallbladder from human and mouse
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PROVIDER: PRJNA639277 | ENA |
REPOSITORIES: ENA
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Project description:Collection of pancreatic islet, pancreas and gallbladder from human and mouse whole transcriptome data.
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Project description:We report here an improved protocol to reprogram mouse gallbladder cells (GBCs) into pancreatic beta cells. To fully understand the extent of reprogramming, mRNA was extracted from FACS-purified MIP-GFP positive insulin-producing cells (namely rGBC2) for RNA-seq after 10-days of in vitro reprogramming. The global gene expression profile of rGBC2 was compared with that of primary gallbladder cells, GBC reprogrammed with the rGBC1 protocol (Hickey et al., 2013) and mouse pancreatic β cells (Benner et al., 2014). We show that rGBC2 from four independent cell batches showed a unique gene expression phenotype. Compared with the rGBC1 protocol, rGBC2 expressed many additional pancreatic β cell genes, suppressed many gallbladder genes and resulted in an expression profile closer to pancreatic β cells.
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Project description:Identification of genes enriched in putative stem/progenitor cells (CD133highPDGFRb- cell population) from the mouse embryonic pancreas that are purified by fluorescence activated cell sorting (FACS). Success in islet transplantation-based therapies for type 1 diabetes mellitus and an extreme shortage of pancreatic islets has motivated efforts to develop renewable sources of islet-replacement tissue. Only a few attempts have been made at prospective isolation of pancreatic stem/progenitor cells, due to the lack of specific markers and the development of cell culture method. This study demonstrates the isolation of pancreatic stem/progenitor cells from the embryonic pancreas by cell sorting. RT-PCR and microarray analysis demonstrated that pancreatic stem/progenitor cells are enriched in CD133highPDGFRb- cell population. During in vivo differentiation, these cell populations have the ability for self-renewal and multipotency, including the formation of insulin-producing cells. Since the strategy is based on the cell sorting using cell surface markers common to human and rodents, it may promote strategies to derive transplantable islet-replacement tissues from human pancreatic stem/progenitor cells. Experiment Overall Design: A couple of total RNAs derived from the different status of PDGFRb- cells was subjected to a dual-color microarray analysis, in which the RNAs from CD133highPDGFRb- cell population was labeled with Cy3 and that of CD133negPDGFRb- cell population with Cy5 respectively.
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2012-07-30 | E-MEXP-3691 | biostudies-arrayexpress
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2015-04-16 | E-GEOD-63020 | biostudies-arrayexpress
Project description:Early postnatal overnutrition causes persistent dysregulation of endocrine pancreas function. We used genome-scale DNA methylation profiling in the suckling-period small litter (SL) mouse model to test whether this occurs via persistent epigenetic changes in pancreatic islets. Although SL islets showed DNA methylation changes directly after weaning and in adulthood, few of these were present at both ages, contrary to our hypothesis. Most interestingly, we discovered that genomic regions that are hypermethylated in exocrine relative to endocrine pancreas tend to gain methylation in islets during aging. Focusing on a subset of genes relevant to β cell function, we showed that these methylation differences are strongly correlated with expression. Together, our results provide the novel insight that DNA methylation changes that occur as islets age indicate an overall epigenetic drift toward the exocrine pancreas epigenome. These concerted shifts in the islet methylome could contribute to the age-associated decline in endocrine pancreas function. Pancreatic islets were isolated from P21/P180 SL or C mice. To ensure purity of islets, 3 rounds of manual picking were performed in each samples. Whole pancreas samples, ~98% of which is exocrine pancreas, were used as exocrine pancreas. There are 5 mice per group.
2014-12-03 | E-GEOD-63811 | biostudies-arrayexpress
Project description:In the past decade, several transcription factors critical for pancreas development have been identified. Despite this success, many of the cell surface and extracellular factors necessary for proper islet morphogenesis and function remain uncharacterized. Previous studies have shown that transgenic over-expression of the transcription factor HNF6 specifically in the pancreatic endocrine cell lineage resulted in the disruption of islet morphogenesis, including dysfunctional endocrine cell sorting, increased islet size, and failure of islets to migrate away from the ductal epithelium. We exploited the dysmorphic islets in pdx1PBHnf6 animals as a tool to identify factors important for islet morphogenesis. Genome-wide microarray analysis was used to identify differences in the gene expression profiles of late gestation and early postnatal pancreas tissue from wild type and pdx1PBHnf6 animals. We report the identification of genes with an altered expression in HNF6 Tg animals and highlight factors with potential importance in islet morphogenesis. Experiment Overall Design: Whole genome microarray analysis was used to identify differences in the gene expression profiles of late gestation and early postnatal pancreas tissue from wild type (WT) animals and HNF6 transgenic animals at a time when the events of normal islet morphogenesis are occurring (e18.5 and postnatal day 1). RNA was isolated from individual pancreata at the appropriate developmental stage. Highly pure samples were pooled according to their genotype (3-5 animals per pool) in order to obtain an adequate quantity of RNA for Affymetrix GeneChip analysis. Twelve samples were run in total, 3 replicates each of 4 groups: embryonic day 18.5, wild type mouse; embryonic day 18.5, HNF6 Transgenic mouse; post-natal day 1, wild type mouse; post-natal day 1, HNF6 Transgenic mouse.
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