Regenerative Medicine Program, Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada.
Institute of Cardiovascular Sciences, Albrechtsen St. Boniface Research Centre, University of Manitoba, Winnipeg, MB, Canada.
Methods Mol Biol. 2024;2835:111-120. doi: 10.1007/978-1-0716-3995-5_10.
The advancement in technology has allowed us to identify and accurately detect new mutations causing genetic disorders. However, their underlying physiological mechanisms of manifestation are not well understood. This chapter is a non-invasive blueprint to how iPSC-based disease modeling can be used to understand the neural activity and provide mechanistic insights for inborn disorder patients with neurological dysfunction seen more prominently with metabolic disorder patients. It has increasingly become easier to create personalized iPSCs from both specific patients and corresponding age and sex-matched controls by using their blood samples. These iPSCs can be used to generate any cell type of the body. This chapter covers how iPSCs can be generated from blood cells and their characterization followed by instructions on differentiating these iPSCs into mature neurons in a petri dish. The chapter most importantly describes how these mature neurons can be evaluated for their activity by using multi-well microelectrode array system and its analysis. This method of generating personalized iPSC derived neurons and their endpoint assessment can be applied to many clinical and preclinical studies. This iPSC-based application can be extrapolated to study any condition which can affect neuronal activity.
技术的进步使我们能够识别和准确检测导致遗传疾病的新突变。然而,它们的表现的潜在生理机制尚不清楚。本章介绍了一种非侵入性的蓝图,展示了如何使用基于 iPSC 的疾病建模来理解神经活动,并为具有神经功能障碍的遗传障碍患者提供机制见解,这些患者在代谢障碍患者中更为明显。现在,通过使用血液样本,越来越容易从特定患者以及相应的年龄和性别匹配的对照者中创建个性化的 iPSC。这些 iPSC 可以用来生成身体的任何细胞类型。本章涵盖了如何从血细胞中生成 iPSC 及其特征描述,以及如何在培养皿中将这些 iPSC 分化为成熟神经元的说明。本章最重要的部分描述了如何使用多井微电极阵列系统及其分析来评估这些成熟神经元的活性。这种生成个性化 iPSC 衍生神经元及其终点评估的方法可应用于许多临床和临床前研究。这种基于 iPSC 的应用可以推广到研究任何可能影响神经元活动的情况。