Transcriptional profiling by 4SU-seq in mouse ESCs and ESC-derived neural progenitor cells.
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ABSTRACT:
PROVIDER: PRJNA475990 | ENA |
REPOSITORIES: ENA
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Project description:Nascent RNA was metabolically labelled with 4SU in undifferentiated and ESC-derived neural progenitor cells (NPCs). 4SU incorporated RNA was isolated and deep-sequenced at day 0 (ESCs), 3, 5 and 7 of differentiation. NPC differentiation was monitored through expression of a GFP reporter insereted into the Sox1 locus (46C reporter ESC line; PMID: 12524553). The aim was to monitor changes in transcription as ESCs differentiate into NPCs and relate this to enhancer activity.
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Project description:Tumor formation constitutes a major obstacle to the clinical application of embryonic stem cells (ESCs). As P-RPCs could successfully integrate into host eyes without development of teratomas or NOG, we sought to identify differentially expressed genes between P-RPCs and ESC-RPCs through genome-wide transcript profiling. Inhibition of Wnt signaling by DKK1 promotes the commitment of ESC-RPCs to more mature retinal cells and reduces the occurrence of NOG to 3%. DKK1-treated ESC-RPCs efficiently integrate to the host retina, form synaptic connections and restore visual function. Here, we report that further differentiation of ESC-derived neural progenitors into retinal progenitor cells (ESC-RPCs) completely eliminates teratomas in ocular transplantation. However, tumor-like neural overgrowth (NOG) occurs in 61% of transplanted eyes.
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Project description:The transplantation of ESCs into living recipients causes teratoma formation which accompanies with several biological reactions in both the transplanted cells and recipient. Co-injection of ESCs with somatic cells that influence donor-recipient homogeneity can affect transcriptional profile of transplanted ESCs. We investigated global gene expression of parental ESCs and ESC-like cells derived from teratomas which were formed by different injection conditions to find transcriptional differences according to donor-recipient homogeneity and somatic cell co-injection. Parental ESCs or teratoma-derived ESC-like cells from different treatments were retrieved for RNA extraction and hybridization on Affymetrix microarrays.
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Project description:Endodermal progenitor cells (EP cells) are derived from human embryonic stem cell(ESC)-derived definitive endoderm (DE) cells. EP cells are cultured in high BMP media and DE cells are in high Activin media. Both cells can be further differentiated to liver, pancreas, etc. We used microarray to detail the global gene expression profile of DE cells and EP cells to delineate the difference of DE cells and EP cells. ESCs were sorted by SSEA3 and SSEA4. DE cells and EP cells were sorted by CXCR4 and CD117. Sorted cells were lysed in RLT buffer. RNA was extracted and hybridized to Affimetrix microarrays. Each group had 3 biological replicates.
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