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从人脱落乳牙干细胞(SHED)来源的诱导多能干细胞分化外周感觉神经元。

Differentiation of peripheral sensory neurons from iPSCs derived from stem cells from human exfoliated deciduous teeth (SHED).

作者信息

Oliveira Nathalia C, Russo Fabiele B, Beltrão-Braga Patricia C B

机构信息

Disease Modeling Laboratory, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.

Neurobiology Laboratory, Scientific Platform Pasteur-USP, São Paulo, Brazil.

出版信息

Front Cell Dev Biol. 2023 Jul 13;11:1203503. doi: 10.3389/fcell.2023.1203503. eCollection 2023.

Abstract

Peripheral nervous system (PNS) sensory alterations are present in several pathologies and syndromes. The use of induced pluripotent stem cell (iPSC) technology is an important strategy to produce sensory neurons in patients who are accomplished in terms of sensory symptoms. The iPSC technology relies on manipulating signaling pathways to resemble what occurs , and the iPSCs are known to carry a transcriptional memory after reprogramming, which can affect the produced cell. To this date, protocols described for sensory neuron production start using iPSCs derived from skin fibroblasts, which have the same ontogenetic origin as the central nervous system (CNS). Since it is already known that the cells somehow resemble their origin even after cell reprogramming, PNS cells should be produced from cells derived from the neural crest. This work aimed to establish a protocol to differentiate sensory neurons derived from stem cells from human exfoliated deciduous teeth (SHED) with the same embryonic origin as the PNS. SHED-derived iPSCs were produced and submitted to peripheral sensory neuron (PSN) differentiation. Our protocol used the dual-SMAD inhibition method, followed by neuronal differentiation, using artificial neurotrophic factors and molecules produced by human keratinocytes. We successfully established the first protocol for differentiating neural crest and PNS cells from SHED-derived iPSCs, enabling future studies of PNS pathologies.

摘要

外周神经系统(PNS)感觉改变存在于多种病理状态和综合征中。诱导多能干细胞(iPSC)技术的应用是在有感觉症状的患者中产生感觉神经元的重要策略。iPSC技术依赖于操纵信号通路以模拟自然发生的情况,并且已知iPSC在重编程后会携带转录记忆,这可能会影响所产生的细胞。迄今为止,描述的感觉神经元产生方案开始使用源自皮肤成纤维细胞的iPSC,其与中枢神经系统(CNS)具有相同的个体发生起源。由于已知即使在细胞重编程后细胞仍在某种程度上类似于其起源,因此PNS细胞应由源自神经嵴的细胞产生。这项工作旨在建立一种方案,以分化来自人脱落乳牙(SHED)的干细胞的感觉神经元,其与PNS具有相同的胚胎起源。产生了SHED衍生的iPSC并进行外周感觉神经元(PSN)分化。我们的方案使用双SMAD抑制方法,随后使用人工神经营养因子和人角质形成细胞产生的分子进行神经元分化。我们成功建立了第一个从SHED衍生的iPSC分化神经嵴和PNS细胞的方案,为未来PNS病理学研究提供了可能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/067c/10374323/f80481a4e9ce/fcell-11-1203503-g001.jpg

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