Heng Boon Chin, Gong Ting, Wang Shuai, Lim Lee Wei, Wu Wutian, Zhang Chengfei
Endodontology, Faculty of Dentistry, University of Hong Kong, Pokfulam, Hong Kong, China; Department of Biological Sciences, Sunway University, Bandar Sunway, Selangor Darul Ehsan, Malaysia.
Endodontology, Faculty of Dentistry, University of Hong Kong, Pokfulam, Hong Kong, China.
J Endod. 2017 Mar;43(3):409-416. doi: 10.1016/j.joen.2016.10.033.
Dental follicle stem cells (DFSCs) possess neurogenic potential because they originate from the embryonic neural crest. This study investigated whether neural differentiation of DFSCs can be enhanced by culture on decellularized matrix substrata (NSC-DECM) derived from neurogenesis of human embryonic stem cells (hESCs).
The hESCs were differentiated into neural stem cells (NSCs), and NSC-DECM was extracted from confluent monolayers of NSCs through treatment with deionized water. DFSCs seeded on NSC-DECM, Geltrex, and tissue culture polystyrene (TCPS) were subjected to neural induction during a period of 21 days. Expression of early/intermediate (Musashi1, PAX6, NSE, and βIII-tubulin) and mature/late (NGN2, NeuN, NFM, and MASH1) neural markers by DFSCs was analyzed at the 7-, 14-, and 21-day time points with quantitative real-time polymerase chain reaction. Immunocytochemistry for detection of βIII-tubulin, PAX6, and NGN2 expression by DFSCs on day 7 of neural induction was also carried out.
Quantitative RT-PCR showed that expression of PAX6, Musashi1, βIII-tubulin, NSE, NGN2, and NFM by DFSCs was enhanced on NSC-DECM versus either the Geltrex or TCPS groups. Immunocytochemistry showed that DFSCs in the NSC-DECM group displayed more intense staining for βIII-tubulin, PAX6, and NGN2 expression, together with more neurite outgrowths and elongated morphology, as compared with either Geltrex or TCPS.
DECM derived from neurogenesis of hESCs can enhance the neurogenic potential of DFSCs.
牙囊干细胞（DFSCs）具有神经源性潜能，因为它们起源于胚胎神经嵴。本研究调查了在源自人类胚胎干细胞（hESCs）神经发生的脱细胞基质底物（NSC-DECM）上培养是否能增强DFSCs的神经分化。
将hESCs分化为神经干细胞（NSCs），并通过用去离子水处理从NSCs的汇合单层中提取NSC-DECM。接种在NSC-DECM、Geltrex和组织培养聚苯乙烯（TCPS）上的DFSCs在21天内进行神经诱导。在第7、14和21天时间点，用定量实时聚合酶链反应分析DFSCs早期/中期（Musashi1、PAX6、NSE和βIII-微管蛋白）和成熟/晚期（NGN2、NeuN、NFM和MASH1）神经标志物的表达。还进行了免疫细胞化学检测神经诱导第7天DFSCs中βIII-微管蛋白、PAX6和NGN2的表达。
定量RT-PCR显示，与Geltrex或TCPS组相比，DFSCs在NSC-DECM上PAX6、Musashi1、βIII-微管蛋白、NSE、NGN2和NFM的表达增强。免疫细胞化学显示，与Geltrex或TCPS相比，NSC-DECM组中的DFSCs对βIII-微管蛋白、PAX6和NGN2表达的染色更强，同时有更多的神经突生长和更长的形态。
源自hESCs神经发生的DECM可增强DFSCs的神经源性潜能。
