Transdifferentiation of human periodontal ligament stem cells into pancreatic cell lineage
Transdifferentiation of human periodontal ligament stem cells into pancreatic cell lineage
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DOI：
10.1002/cbf.3057
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发表时间：
2014-10-01
影响因子：
3.6
通讯作者：
Heo, Jung Sun
中科院分区：
文献类型：
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作者：
Lee, Jeong Seok;An, Seong Yeong;Heo, Jung Sun
人牙周膜来源的干细胞（PDLSC）表现出自我更新能力和多谱系分化潜力。在这项研究中，我们研究了人 PDLSC 向胰岛细胞的转分化潜力。为了形成三维 (3D) 簇，PDLSC 在含有分化诱导剂的基质胶中培养。我们发现培养 6 天后，PDLSC 经历了类似于胰岛样细胞簇 (ICC) 的形态变化。使用扫描电子显微镜分析进一步评估 PDLSC 衍生的 ICC 的形态特征。通过逆转录聚合酶链反应分析，我们发现与未分化的PDLSC相比，PDLSC衍生的ICC中多能性基因下调，而早期内胚层和胰腺分化基因上调。此外，我们发现PDLSC衍生的ICC能够响应高浓度葡萄糖而分泌胰岛素，验证了它们向胰岛细胞的功能分化。最后，我们还进行了双硫腙染色，以及胰腺分化标记物的免疫荧光测定和荧光激活细胞分选分析，以确认 PDLSC 衍生的 ICC 的分化状态。这些结果表明 PDLSC 可以转分化为功能性胰岛样细胞，并为胰腺修复提供新型替代细胞群。版权所有 (c) 2014 约翰·威利父子有限公司
Human periodontal ligament-derived stem cells (PDLSCs) demonstrate self-renewal capacity and multilineage differentiation potential. In this study, we investigated the transdifferentiation potential of human PDLSCs into pancreatic islet cells. To form three-dimensional (3D) clusters, PDLSCs were cultured in Matrigel with media containing differentiation-inducing agents. We found that after 6days in culture, PDLSCs underwent morphological changes resembling pancreatic islet-like cell clusters (ICCs). The morphological characteristics of PDLSC-derived ICCs were further assessed using scanning electron microscopy analysis. Using reverse transcription-polymerase chain reaction analysis, we found that pluripotency genes were downregulated, whereas early endoderm and pancreatic differentiation genes were upregulated, in PDLSC-derived ICCs compared with undifferentiated PDLSCs. Furthermore, we found that PDLSC-derived ICCs were capable of secreting insulin in response to high concentrations of glucose, validating their functional differentiation into islet cells. Finally, we also performed dithizone staining, as well as immunofluorescence assays and fluorescence-activated cell sorting analysis for pancreatic differentiation markers, to confirm the differentiation status of PDLSC-derived ICCs. These results demonstrate that PDLSCs can transdifferentiate into functional pancreatic islet-like cells and provide a novel, alternative cell population for pancreatic repair. Copyright (c) 2014 John Wiley & Sons, Ltd.