Spontaneous differentiation of periodontal ligament stem cells into myofibroblast during ex vivo expansion
Spontaneous differentiation of periodontal ligament stem cells into myofibroblast during ex vivo expansion
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DOI：
10.1002/jcp.28639
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发表时间：
2019-11-01
影响因子：
5.6
通讯作者：
Morita, Ikuo
中科院分区：
文献类型：
--
作者：
Iwasaki, Kengo;Komaki, Motohiro;Morita, Ikuo
牙周炎的特征是牙齿支持组织的慢性炎症和破坏。牙周膜干细胞（periodicalligamentstemcell，PDLSC）是从牙周膜中分离的间充质干细胞（mesenchymalstemcell，MSC），是牙周组织再生的关键组织。尽管PDLSC的移植被提出作为新的再生疗法，但关于PDLSC在离体扩增期间的特征变化的信息有限。本研究以PDLSCs在标准细胞培养过程中发生的形态学变化为研究对象，旨在探讨PDLSCs干细胞特性的变化，寻找维持干细胞特性的培养条件。采用体外成骨细胞和脂肪细胞分化的方法检测PDLSCs的特性。肌成纤维细胞的分化证实了免疫组织化学和胶原凝胶收缩试验。通过β-gal染色检查复制衰老。PDLSC在体外扩增过程中形态从梭形变为扁平和宽形。形态学改变后，PDLSC显示出肌成纤维细胞的几个特征，包括广泛的应力纤维形成、收缩活性和肌成纤维细胞标志物表达。形态学改变后，成骨细胞和脂肪细胞分化能力降低，干细胞相关基因表达减少。β-Gal染色与PDLSCs形态学改变不一定相关。此外，外源性添加bFGF和PDGF-BB有助于维持PDLSCs的梭形和成骨分化潜能。本研究证实了PDLSCs在体外扩增过程中的自发分化，为PDLSCs的临床应用提供了细胞培养条件的重要信息。
Periodontitis is characterized by the chronic inflammation and destruction of tooth-supporting tissues. Periodontal ligament stem cell (PDLSC) is the mesenchymal stem cell (MSC) population isolated from periodontal ligament, which is the key tissue for regeneration of periodontal tissues. Although transplantation of PDLSCs is proposed as novel regenerative therapy, limited information is available, regarding the characteristic change of PDLSCs during ex vivo expansion. In this study, we encountered morphological change of PDLSCs during standard cell culture and aimed to investigate the change of PDLSCs in stem cell characteristics and to search for the culture condition to maintain stem cell properties. Characteristics of PDLSCs were examined using in vitro osteoblast and adipocyte differentiation. Myofibroblast differentiation was confirmed using immunohistochemistry and collagen gel contraction assay. Replicative senescence was examined by beta-gal staining. PDLSCs changed their morphology from spindle to flat and wide during ex vivo expansion. After the morphological change, PDLSCs showed several features of myofibroblast including extensive stress fiber formation, contraction activity, and myofibroblast marker expression. Upon the morphological change, osteoblastic and adipocyte differentiation capacity were reduced and expression of stem cell-related genes were decreased. beta-Gal staining was not always correlated with the morphological change of PDLSCs. Moreover, exogenous addition of bFGF and PDGF-BB served to maintain spindle shape and osteoblastic differentiation potential of PDLSCs. This study demonstrates that spontaneous differentiation of PDLSCs during ex vivo expansion and may provide the important information of cell culture condition of PDLSCs for clinical use.