Neural differentiation of mouse embryonic stem cells studied by FTIR spectroscopy
Neural differentiation of mouse embryonic stem cells studied by FTIR spectroscopy
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DOI：
10.1016/j.molstruc.2010.01.007
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发表时间：
2010-04-01
影响因子：
3.8
通讯作者：
Heraud, Philip
中科院分区：
文献类型：
--
作者：
Tanthanucha, Waraporn;Thumanu, Kanjana;Heraud, Philip
胚胎干细胞衍生的神经细胞（ESNCs）具有作为神经元来源的潜力，用于未来治疗脑肿瘤和其他神经系统疾病和障碍的细胞疗法。神经细胞类型的分选被认为是这些细胞在中枢神经系统（CNS）的基于细胞的疗法中的临床应用的最重要的处理之一。本研究采用实验室FTIR和同步辐射FTIR（SR-FTIR）显微光谱技术，对小鼠胚胎干细胞（mESCs）分化为不同类型神经细胞的FTIR标记物进行鉴定。主成分分析（PCA）和无监督层次聚类分析（UHCA）显示能够将mESCs的发育阶段区分为三种细胞类型：胚状体（EB）、神经祖细胞（NPC）和ESNCs。此外，PCA提供了识别潜在FTIR“标记带”的手段，这些标记带在干细胞沿着神经谱系分化过程中发生了巨大变化。这些似乎与脂质（来自CH 2和CH 3伸缩振动的谱带，位于2959、2923和2852 cm（-1）附近）和蛋白质（酰胺I谱带的变化，位于1659和1637 cm（-1）附近）的变化有关。结果表明，随着分化时间的推移，细胞的脂质含量显著增加，表明甘油磷脂的表达增加。酰胺I谱的变化表明，随着mESC向ESNCs分化，富含α-螺旋的蛋白质随之增加，富含β-折叠的蛋白质相应减少，这与可能与神经结构和功能的建立有关的细胞骨架蛋白的变化相对应。皇冠版权所有（C）2010由爱思唯尔B. V.出版保留所有权利。
Embryonic Stem-derived Neural Cells (ESNCs) hold potential as a source of neurons for a cell-based therapy for the treatment of brain tumors, and other neurological diseases and disorders in the future. The sorting of neural cell types is envisaged to be one of the most important processed for clinical application of these cells in cell-based therapies of the central nervous system (CNS). In this study, laboratory-based FTIR and Synchrotron-FTIR (SR-FTIR) microspectroscopy were used to identify FTIR marker for distinguishing different neural cell types derived from the differentiation of mouse embryonic stem cells (mESCs). Principal Component Analysis (PCA) and Unsupervised Hierarchical Cluster Analysis (UHCA) were shown to be able to distinguish the developmental stage of mESCs into three cell types: embryoid bodies (EBs), neural progenitor cells (NPCs), and ESNCs. Moreover, PCA provided the mean for identifying potential FTIR "marker bands" that underwent dramatic changes during stem cell differentiation along neural lineages. These appeared to be associated with changes in lipids (bands from CH2 and CH3 stretching vibrations at similar to 2959, 2923 and 2852 cm(-1)) and proteins (changes in the amide I band at similar to 1659 and 1637 cm(-1)). The results suggested that lipid content of cells increased significantly over the time of differentiation, suggesting increased expression of glycerophospholipids. Changes in the amide I profile, suggested concomitant increases in alpha-helix rich proteins as mESCs differentiated towards ESNCs, with a corresponding decrease in beta-sheet rich proteins, corresponding with changes in cytoskeleton protein which may have been taking place involved with the establishment of neural structure and function. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.