Zuccoli Giuliana S, Martins-de-Souza Daniel, Guest Paul C, Rehen Stevens K, Nascimento Juliana Minardi
Lab of Neuroproetomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, SP, 13083-970, Brazil.
UNICAMP's Neurobiology Center, Campinas, Brazil.
Adv Exp Med Biol. 2017;974:279-287. doi: 10.1007/978-3-319-52479-5_26.
The mechanisms underlying the pathophysiology of psychiatric disorders are still poorly known. Most of the studies about these disorders have been conducted on postmortem tissue or in limited preclinical models. The development of human induced pluripotent stem cells (iPSCs) has helped to increase the translational capacity of molecular profiling studies of psychiatric disorders through provision of human neuronal-like tissue. This approach consists of generation of pluripotent cells by genetically reprogramming somatic cells to produce the multiple neural cell types as observed within the nervous tissue. The finding that iPSCs can recapitulate the phenotype of the donor also affords the possibility of using this approach to study both the disease and control states in a given medical area. Here, we present a protocol for differentiation of human pluripotent stem cells to neural progenitor cells followed by subcellular fractionation which allows the study of specific cellular organelles and proteomic analysis.
精神疾病病理生理学的潜在机制仍知之甚少。关于这些疾病的大多数研究都是在死后组织或有限的临床前模型中进行的。人类诱导多能干细胞(iPSC)的发展通过提供类人神经组织,有助于提高精神疾病分子谱研究的转化能力。这种方法包括通过对体细胞进行基因重编程来产生多能细胞,以产生神经组织中观察到的多种神经细胞类型。iPSC能够重现供体表型这一发现,也为使用这种方法研究特定医学领域的疾病和对照状态提供了可能性。在此,我们展示了一种将人类多能干细胞分化为神经祖细胞,随后进行亚细胞分级分离的方案,该方案可用于研究特定细胞器并进行蛋白质组分析。