Zeng Sujuan, Zhao Xuedan, Zhang Lingling, Pathak Janak L, Huang Wenyan, Li Yunyang, Guan Hongbing, Zhao Wanghong, Ge Lihong, Shu Yan
Department of Pediatric Dentistry, Affiliated Stomatology Hospital of Guangzhou Medical University, Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou, 510182, China.
GuangDong Second Traditional Chinese Medicine Hospital, Guangzhou, 510095, China.
J Biol Eng. 2020 Dec 9;14(1):29. doi: 10.1186/s13036-020-00251-4.
The stem cells of human exfoliated deciduous teeth (SHEDs) are considered to be one of the main sources of seed cells in stem cell therapy. The aim of this study was to examine the effect of ciliary neurotrophic factor (CNTF) on neurogenic differentiation of SHEDs. With the consent of parents, SHEDs from 6 to 8 year old children were isolated and cultured. The mesenchymal stemness and the potential of multidirectional (adipogenic and osteogenic) differentiation for the isolated SHEDs were firstly determined. The effect of CNTF on specific neurogenic differentiation of SHEDs was then examined by detecting the expression of marker genes and proteins via RT-PCR, immunoblotting, and immunofluorescence microscopy. The isolated SHEDs expressed specific surface markers of mesenchymal stem cells, and their potential of osteogenic and adipogenic differentiation were confirmed. CNTF promoted the differentiation of SHEDs into neuron-like cells with a high expression of acetylcholine transferase (CHAT), a marker of cholinergic neurons. The expression of other neuron markers including nestin, microtubule-associated protein 2 (MAP 2), and β-tublin III was also detected. Interestingly, the expression of neurogenic markers was maintained at a high level after neurogenic induction. SHEDs can be induced by CNTF to differentiate into cholinergic neuron-like cells under appropriate culture conditions. Our findings have laid a foundation for future use of SHEDs to treat neurological diseases.
人乳牙脱落干细胞（SHEDs）被认为是干细胞治疗中种子细胞的主要来源之一。本研究的目的是检测睫状神经营养因子（CNTF）对SHEDs神经源性分化的影响。在获得家长同意后，分离并培养了6至8岁儿童的SHEDs。首先确定了分离出的SHEDs的间充质干性以及多向（成脂和成骨）分化潜能。然后通过逆转录聚合酶链反应（RT-PCR）、免疫印迹和免疫荧光显微镜检测标记基因和蛋白质的表达，来研究CNTF对SHEDs特定神经源性分化的影响。分离出的SHEDs表达间充质干细胞的特异性表面标志物，并证实了它们的成骨和成脂分化潜能。CNTF促进SHEDs分化为具有高表达乙酰胆碱转移酶（CHAT，胆碱能神经元的标志物）的神经元样细胞。还检测了包括巢蛋白、微管相关蛋白2（MAP 2）和β-微管蛋白III在内的其他神经元标志物的表达。有趣的是，神经源性诱导后神经源性标志物的表达维持在高水平。在适当的培养条件下，SHEDs可被CNTF诱导分化为胆碱能神经元样细胞。我们的研究结果为未来使用SHEDs治疗神经疾病奠定了基础。
