Chromatin accessibility in canine stromal cells and its implications for canine somatic cell reprogramming.

Stem Cells Translational Medicine
Maria QuestaAmir Kol

Abstract

Naturally occurring disease in pet dogs is an untapped and unique resource for stem cell-based regenerative medicine translational research, given the many similarities and complexity such disease shares with their human counterparts. Canine-specific regulators of somatic cell reprogramming and pluripotency maintenance are poorly understood. While retroviral delivery of the four Yamanaka factors successfully reprogrammed canine embryonic fibroblasts, adult stromal cells remained resistant to reprogramming in spite of effective viral transduction and transgene expression. We hypothesized that adult stromal cells fail to reprogram due to an epigenetic barrier. Here, we performed assay for transposase-accessible chromatin using sequencing (ATAC-seq) on canine stromal and pluripotent stem cells, analyzing 51 samples in total, and establishing the global landscape of chromatin accessibility before and after reprogramming to induced pluripotent stem cells (iPSC). We also studied adult stromal cells that do not yield iPSC colonies to identify potential reprogramming barriers. ATAC-seq analysis identified distinct cell type clustering patterns and chromatin remodeling during embryonic fibroblast reprogramming. Compared with embryonic f...Continue Reading

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Methods Mentioned

BETA
Assay
PCR
reverse transcription PCR
Transfection

Software Mentioned

HTStream
Cluster
Bioconductor package
Ensembl
GraphPad Prism
voom Bioconductor
ChIPseeker
Java
TreeView
deeptools

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