Chie Stefanie Elke, Szentpetery Zsofia, Generali Melanie, Kuhlmann Tanja, Natalucci Giancarlo, Miletta Maria Consolata
Larsson-Rosenquist Foundation Center for Neurodevelopment, Growth and Nutrition of the Newborn, Department of Neonatology, University of Zurich and University Hospital Zurich, Zurich, Switzerland.
ZNZ PhD program, University of Zurich, Zurich, Switzerland.
Bio Protoc. 2025 May 5;15(9):e5227. doi: 10.21769/BioProtoc.5227.
Neurons and oligodendrocytes are the building blocks of the brain. Neurons form synaptic connections and transmit signals, while oligodendrocytes, including oligodendrocyte precursor cells (OPCs) and their derivatives, are vital for central nervous system maintenance and myelination. The demand for human-specific neuron-oligodendrocyte model systems to study these interactions has grown, yet co-culture protocols remain limited. Recent advancements in the field provide methods for deriving co-cultures of neurons and OPCs from human induced pluripotent stem cells (hiPSC), each with distinct benefits and challenges. This study presents a time-efficient, reproducible method to derive neurons and O4-expressing oligodendrocytes, followed by a straightforward co-culture system that minimizes astrocyte differentiation and ensures robust neuron and oligodendrocyte populations. Key features • Reliable, stable generation of neurons and O4-expressing oligodendrocytes within a practical timeframe. • Co-culture system utilizing hIPSC-derived neurons and O4-expressing oligodendrocytes. • Maturation of neurons and oligodendrocytes achieved within 10 days of co-culturing. Graphical overview The diagram outlines the sequential steps involved in the preparation, differentiation, and analysis phases. Key stages include the differentiation of neural progenitor cells (NPCs) into O4-expressing oligodendrocytes and neurons separately and then combining them into a co-culture, which can then be used for further experiments.
神经元和少突胶质细胞是大脑的组成部分。神经元形成突触连接并传递信号，而少突胶质细胞，包括少突胶质前体细胞（OPC）及其衍生物，对于中枢神经系统的维持和髓鞘形成至关重要。研究这些相互作用的人类特异性神经元-少突胶质细胞模型系统的需求不断增加，但共培养方案仍然有限。该领域的最新进展提供了从人类诱导多能干细胞（hiPSC）中获得神经元和OPC共培养物的方法，每种方法都有不同的优点和挑战。本研究提出了一种高效、可重复的方法来获得神经元和表达O4的少突胶质细胞，随后是一个简单的共培养系统，该系统可最大限度地减少星形胶质细胞分化，并确保强大的神经元和少突胶质细胞群体。关键特性 • 在实际时间范围内可靠、稳定地生成神经元和表达O4的少突胶质细胞。 • 利用hIPSC衍生的神经元和表达O4的少突胶质细胞的共培养系统。 • 共培养10天内实现神经元和少突胶质细胞的成熟。图形概述 该图概述了制备、分化和分析阶段所涉及的顺序步骤。关键阶段包括将神经祖细胞（NPC）分别分化为表达O4的少突胶质细胞和神经元，然后将它们组合成共培养物，然后可用于进一步实验。
