Unknown,Transcriptomics,Genomics,Proteomics

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Cells with Surface Expression of CD133highCD71low are Enriched for Colony-Forming Progenitor Cells in the Adult Murine Pancreas

ABSTRACT: Previously, we identified tripotent colony-forming progenitor cells in the adult murine pancreas that were capable of self-renewal and differentiation into duct, acinar and endocrine cells in vitro. However, the incidence of these progenitor cells was low (~1% among pancreatic cells in adult 2-4 month-old mice). The goal of the current study was to find cell-surface markers that can distinguish and enrich pancreatic colony-forming progenitor cells. It was found that pancreatic cells could be divided into CD133+CD71- , CD133highCD71low and CD133lowCD71low and CD133-CD71- cells. CD133highCD71low cells, but not other cell populations, were the most enriched for colony-forming progenitors. Interestingly, both CD133+CD71- and CD133highCD71low cells expressed ductal markers as shown by gene expression RT-PCR analysis. To further characterize CD133+CD71- (designated as R1 cells) and CD133highCD71low (designated as R2 cells) ductal cell populations, genome-wide gene expression analysis using RNA-seq was performed. The genes that were differentially expressed by R1 and R2 cells are deposited here. Subsequent pathway analysis of the RNA-seq data revealed that R2 and R1 cells have propensity for cell migration and immune response, respectively. These gene expression analyses suggested ductal cells of the adult pancreas are heterogeneous in nature. Two populations of cells are sorted from the pancreas of adult 2-4 month old C57Bl/6 mice and their RNA profiles are determined. These cell populations are CD133(high)CD71(low) cells (3 samples) and CD133(positive)CD71(negative) (2 samples).

ORGANISM(S): Mus musculus

SUBMITTER: Xiwei Wu

PROVIDER: E-GEOD-72990 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:It is unclear how epigenetic changes regulate the induction of erythroid-specific genes during terminal erythropoiesis. Here we use global mRNA sequencing (mRNA-seq) and chromatin immunoprecipitation coupled to high-throughput sequencing (CHIP-seq) to investigate the changes that occur in mRNA levels, RNA Polymerase II (Pol II) occupancy and multiple post-translational histone modifications when erythroid progenitors differentiate into late erythroblasts. Among genes induced during this developmental transition, there was an increase in the occupancy of Pol II, the activation marks H3K4me2, H3K4me3, H3K9Ac and H4K16Ac, and the elongation methylation mark H3K79me2. In contrast, genes that were repressed during differentiation showed relative decreases in H3K79me2 levels yet had levels of Pol II binding and active histone marks similar to those in erythroid progenitors. We also found that relative changes in histone modification levels-in particular, H3K79me2 and H4K16ac-were most predictive of gene expression patterns. Our results suggest that in terminal erythropoiesis both promoter and elongation-associated marks contribute to the induction of erythroid genes, while gene repression is marked by changes in histone modifications mediating Pol II elongation. Our data maps the epigenetic landscape of terminal erythropoiesis and suggests that control of transcription elongation regulates gene expression during terminal erythroid differentiation. Mouse fetal liver cells are double-labeled for erythroid-specific TER119 and non erythroid-specific transferrin receptor (CD71) and then sorted by flow-cytometry. E14.5 fetal livers contain at least five distinct populations of cells (R1 through R5); as they progressively differentiate they gain TER119 and then gain and subsequently lose CD71. CFU-E cells and proerythroblasts make up the R1 population; R2 consists of proerythroblasts and early basophilic erythroblasts; R3 includes early and late basophilic erythroblasts; R4 is mostly polychromatophilic and orthochromatophilic erythroblasts; and R5 is comprised of late orthochromatophilic erythroblasts and reticulocytes. We have sorted for R2-R5 cells for RNA-seq experiment.

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Cells with Surface Expression of CD133highCD71low are Enriched for Colony-Forming Progenitor Cells in the Adult Murine Pancreas

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