Project description:This experiment used RNA-Seq technology to explore gene expression in mouse Insm1^GFP/Pdx1^CFP HIGH [het/het] FACS sorted pancreatic cells (pre-beta cells) and Insm1^GFP/Pdx1^CFP LOW [het/het] cells (other endocrine progenitors) at E15.5 and E18.5. Comparison of Insm1 +/Pdx HIGH and Insm1 +/Pdx LOW cells revealed a set of differentially expressed genes that are required for beta cell specification.
Project description:This experiment used RNA-Seq technology to examine transcription profile in FACS-sorted MIP-EGFP+ mouse pancreatic cells at E16.5 (nascent beta cells) and P60 (mature beta cells). Such an analysis should reveal the gene transcription alterations for beta cell development and for function.
Project description:This experiment used RNA-Seq technology to explore gene expression in mouse Ngn3^GFP/+ [het] FACS sorted pancreatic cells at E15.5 (commited endocrine progenitor cells) and in Ngn3^GFP/GFP [null] at E15.5 (defective endocrine progenitor cells). This experiment is designed to understand the gene expression alteration in the endocrine lineage at different embryonic days. The aim is to understand both Ngn3 dependent and independent gene expression profiles so as to reveal the instructive signals that specfy the collective endocrine islet cell fate or specific islet cell type.
Project description:Aim: To determine the baseline gene expression profile of human gallbladder and cystic duct cells at various stages from isolation in fresh biopsy samples to early and later passages in culture.
Project description:Knowledge of the genetic signatures of specific pancreatic cellular populations, both during development and in the adult animal, provides an objective basis for evaluating genetic signatures of cultured human embryonic stem cells (hESCs) and reprogrammed pancreatic mouse cells. Previous transcriptome profiling studies have utilized DNA microarrays, which have well-defined limitations, and few have utilized FACS-purified cell populations. To overcome both of these limitations we utilized a series of mice with fluorescent reporter alleles (Sox17, Pdx1, Ptf1a, Ngn3, Insm1) or transgene (Ins1) to isolate three biological replicates of 12 different pancreatic progenitor or adult cell populations between embryonic day 8.0 and post-natal day 60. These RNAs were then used to make 36 libraries which were sequenced to an average depth of 50 million reads each. After mapping these reads to mouse genome using a standardized analysis pipeline the resulting datasets were analyzed in several ways.
Project description:The stage at which beta cells become functional were first determined with GSIS and Ca++ imaging. Then beta cells at key stages of maturation were purified and their gene expression alterations were determined with RNAseq and exhaustive bioinformatics analyses.
Project description:Mouse gall bladder cells from MIP-GFP mice were expanded in vitro and transduced with adenoviral vectors expressing the transcription factors Neurog3, Pdx1 and MafA. Reprogrammed GFP+ cells were then FACS-sorted for RNA isolation.
Project description:Islet β-cells from newborn mammals need a maturation process to become mature functional beta cells. The detailed molecular mechanisms were not completely understood. This study was designed to reveal the dynamic gene expression changes during pancreatic beta-cell maturation in postnatal mice. We also want to understand how genetic mutations that impair beta-cell function change the genetic networks involved in the beta-cell maturation process. For these aims, pancreatic beta cells were isolated at P1 islets based on the expression of a MipeGFP transgene in a genetic background with pancreatic specific inactivation of Myt1, Myt1L, and St18 (denoted as MytDelpanc; MipeGFP).