Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations.
Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations.
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DOI：
10.1371/journal.pone.0183358
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Fricker RA
中科院分区：
文献类型：
--
作者：
Griffin SM;Pickard MR;Orme RP;Hawkins CP;Williams AC;Fricker RA
维生素B3已被证明在胚胎发育过程中发挥重要作用。具体而言，越来越多的证据表明，烟酰胺，维生素B3的生物活性形式，在干细胞分化为成熟细胞表型（包括中枢神经系统（CNS）的表型）中起着关键作用。详细了解小分子在神经元分化过程中的作用不仅对于揭示谱系特化的潜在机制至关重要，而且对于建立更有效的分化方案以获得用于神经退行性疾病如亨廷顿病（HD）的再生疗法的临床相关细胞也至关重要。因此，本研究旨在研究烟酰胺促进干细胞转化为成熟CNS神经元的潜力。在小鼠胚胎干细胞（mESC; Sox 1GFP敲入46 C细胞系）向神经命运转化期间，将烟酰胺用于分化小鼠胚胎干细胞。使用免疫细胞化学和形态学分析方法评估细胞增殖、分化和成熟的变化。结果表明，当在分化的初始阶段加入10 mM烟酰胺时，可促进贴壁单层培养物中ESC向神经谱系的加速进展。到体外（DIV）14天，早期暴露于烟酰胺显示出增加了分化的β III-微管蛋白阳性神经元的数量。烟酰胺降低了多能干细胞的比例，同时在4 DIV时增加了神经祖细胞的数量。然后，这些祖细胞经历快速转化为神经元，通过在14 DIV时培养物中Sox 1表达的减少和神经祖细胞数量的减少来观察。此外，在烟酰胺存在下产生的GABA能神经元在14 DIV时显示出神经突的成熟度和复杂性增加。因此，单独添加烟酰胺导致多能细胞加速传代通过谱系特化并进一步达到非分裂成熟神经元。我们的研究结果表明，在最佳剂量范围内，烟酰胺能够单独和选择性地指导胚胎干细胞向成熟神经元的转化，因此可能是正常脑发育的关键因素，从而支持维生素及其代谢产物在早期CNS发育过程中的基本作用的先前证据。此外，烟酰胺可以提供一种简单有效的补充剂，以增强干细胞向临床相关神经元的转化。
Vitamin B3 has been shown to play an important role during embryogenesis. Specifically, there is growing evidence that nicotinamide, the biologically active form of vitamin B3, plays a critical role as a morphogen in the differentiation of stem cells to mature cell phenotypes, including those of the central nervous system (CNS). Detailed knowledge of the action of small molecules during neuronal differentiation is not only critical for uncovering mechanisms underlying lineage-specification, but also to establish more effective differentiation protocols to obtain clinically relevant cells for regenerative therapies for neurodegenerative conditions such as Huntington’s disease (HD). Thus, this study aimed to investigate the potential of nicotinamide to promote the conversion of stem cells to mature CNS neurons. Nicotinamide was applied to differentiating mouse embryonic stem cells (mESC; Sox1GFP knock-in 46C cell line) during their conversion towards a neural fate. Cells were assessed for changes in their proliferation, differentiation and maturation; using immunocytochemistry and morphometric analysis methods. Results presented indicate that 10 mM nicotinamide, when added at the initial stages of differentiation, promoted accelerated progression of ESCs to a neural lineage in adherent monolayer cultures. By 14 days in vitro (DIV), early exposure to nicotinamide was shown to increase the numbers of differentiated βIII-tubulin-positive neurons. Nicotinamide decreased the proportion of pluripotent stem cells, concomitantly increasing numbers of neural progenitors at 4 DIV. These progenitors then underwent rapid conversion to neurons, observed by a reduction in Sox 1 expression and decreased numbers of neural progenitors in the cultures at 14 DIV. Furthermore, GABAergic neurons generated in the presence of nicotinamide showed increased maturity and complexity of neurites at 14 DIV. Therefore, addition of nicotinamide alone caused an accelerated passage of pluripotent cells through lineage specification and further to non-dividing mature neurons. Our results show that, within an optimal dose range, nicotinamide is able to singly and selectively direct the conversion of embryonic stem cells to mature neurons, and therefore may be a critical factor for normal brain development, thus supporting previous evidence of the fundamental role of vitamins and their metabolites during early CNS development. In addition, nicotinamide may offer a simple effective supplement to enhance the conversion of stem cells to clinically relevant neurons.
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影响因子：
2.6
作者：
Giammona, Lisa M.;Panuganti, Swapna;Kemper, Jan M.;Apostolidis, Pani A.;Lindsey, Stephan;Papoutsakis, Eleftherios T.;Miller, William M.
通讯作者：
Miller, William M.
影响因子：
9.8
作者：
Lowell, Sally;Benchoua, Alexandra;Heavey, Barry;Smith, Austin G
通讯作者：
Smith, Austin G
影响因子：
9.8
作者：
Chatzi, Christina;Brade, Thomas;Duester, Gregg
通讯作者：
Duester, Gregg
影响因子：
5.2
作者：
Nefzger, Christian M.;Su, Colin T.;Pouton, Colin W.
通讯作者：
Pouton, Colin W.
影响因子：
3.4
作者：
Anderson, D. W.;Bradbury, K. A.;Schneider, J. S.
通讯作者：
Schneider, J. S.