title: Involvement of gene methylation changes in the differentiation of human amniotic epithelial cells into islet-like cell clusters.
abstract: Insulin-dependent diabetes results from destruction of the insulin-producing β-cells of the pancreas. Islet cell transplantation is a promising cure for diabetes. Here, we induced human amniotic epithelial cells (hAECs) to differentiate into islet-like cell clusters by nicotinamide plus betacellulin in vitro, and further investigated the DNA methylation status by a Nimble MeDIP microarray before and after cell differentiation to shed light on the molecular mechanisms of this differentiation. In addition, 5-Aza-2'-deoxycytidine was used to investigate whether the differentiation of hAECs into islet-like cells occurred through demethylation. Purified hAECs (CK18(+)/E-cadherin(+)/CD29(+)/CD90(-)/CD34(-)/CD45(-)) were isolated from human amnia. After induction, hAECs were found to be insulin positive and sensitive to glucose, indicating successful induction to islet-like cells. The methylation status of cell cytoskeleton-related genes was down-regulated and that of negative regulation of cell adhesion-related genes was up-regulated. The methylation status of pancreas development-related genes such as HNF1α and DGAT1 was decreased in hAECs after induction. After brief demethylation, INS gene expression was up-regulated in islet-like cell clusters, suggesting that DNA methylation changes were associated with the differentiation of hAECs into islet-like cell clusters.
authors.affiliation: 1 Institute of Human Reproduction and Stem Cell Engineering, Central South University , Changsha, People's Republic of China .
authors.forename: Lin
authors.initials: L
authors.lastname: Peng
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Guangxiu
authors.initials: G
authors.lastname: Lu
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D019175:DNA Methylation; D004847:Epithelial Cells / Q000502:physiology*; D015870:Gene Expression; D063990:Gene Ontology; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D018874:Spheroids, Cellular / Q000378:metabolism / Q000382:microbiology*