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PubMed · 39821804

A Protocol for Detecting DNA Methylation Changes at CpG Sites of Stemness-Related Genes in Aging Stem Cells.

Mahshid Hodjat·Seyed Mojtaba Daghighi·Mohammad Abdollahi
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Abstract

Aging adversely affects the self-renewal and differentiation capabilities of stem cells, which impairs tissue regeneration as well as the homeostasis. Epigenetic mechanisms, specifically DNA methylation, play a key role in the maintenance of pluripotency in stem cells and regulation of pluripotency-related gene expression. Age-related modifications in methylation patterns could influence the expression of genes critical for stem cell potency maintenance, including transcription factors Nanog and Sox2. The following chapter describes a step-by-step bisulfite sequencing protocol for detection of methylation changes in the aging stem cells and provides valuable insights into the stem cells epigenetic profile. Further, the methodology describes the steps of genomic DNA extraction, bisulfite conversion, real-time PCR amplification, and sequencing for an in-depth view of the epigenetic profile derived from aging stem cells.

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DNA MethylationCpG IslandsHumansEpigenesis, GeneticStem CellsCellular SenescenceAgingReal-Time Polymerase Chain ReactionSequence Analysis, DNASulfitesAnimalsSOXB1 Transcription FactorsAging stem cellsBisulfite sequencingDNA methylationEpigeneticsNanog homeobox proteinProtocolSox2Journal Article

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Mahshid Hodjat, Seyed Mojtaba Daghighi, Mohammad Abdollahi. 2025. A Protocol for Detecting DNA Methylation Changes at CpG Sites of Stemness-Related Genes in Aging Stem Cells.. https://doi.org/10.1007/7651_2024_590

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