Dooves Stephanie, Nadadhur Aishwarya G, Gasparotto Lisa, Heine Vivi M
Pediatric Neurology, Emma Children's Hospital, Amsterdam UMC, Amsterdam, Neuroscience, Vrije Universiteit Amsterdam, The Netherlands.
Department of Functional Genomics, Center for Neurogenomics and Cognitive Research, Amsterdam Neuroscience, Vrije Universiteit Amsterdam, The Netherlands.
Bio Protoc. 2019 Sep 5;9(17):e3350. doi: 10.21769/BioProtoc.3350.
Crosstalk between neurons and oligodendrocytes is important for proper brain functioning. Multiple co-culture methods have been developed to study oligodendrocyte maturation, myelination or the effect of oligodendrocytes on neurons. However, most of these methods contain cells derived from animal models. In the current protocol, we co-culture human neurons with human oligodendrocytes. Neurons and oligodendrocyte precursor cells (OPCs) were differentiated separately from pluripotent stem cells according to previously published protocols. To study neuron-glia cross-talk, neurons and OPCs were plated in co-culture mode in optimized conditions for additional 28 days, and prepared for OPC maturation and neuronal morphology analysis. To our knowledge, this is one of the first neuron-OPC protocols containing all human cells. Specific neuronal abnormalities not observed in mono-cultures of Tuberous Sclerosis Complex (TSC) neurons, became apparent when TSC neurons were co-cultured with TSC OPCs. These results show that this co-culture system can be used to study human neuron-OPC interactive mechanisms involved in health and disease.
神经元与少突胶质细胞之间的相互作用对大脑的正常功能至关重要。人们已经开发出多种共培养方法来研究少突胶质细胞的成熟、髓鞘形成或少突胶质细胞对神经元的影响。然而，这些方法大多包含来自动物模型的细胞。在本实验方案中，我们将人类神经元与人类少突胶质细胞进行共培养。神经元和少突胶质前体细胞（OPCs）根据先前发表的方案分别从多能干细胞分化而来。为了研究神经元与神经胶质细胞之间的相互作用，将神经元和OPCs以共培养模式接种在优化条件下，再培养28天，并为OPC成熟和神经元形态分析做准备。据我们所知，这是首批包含所有人类细胞的神经元-OPC实验方案之一。当结节性硬化症（TSC）神经元与TSC OPCs共培养时，在TSC神经元单培养中未观察到的特定神经元异常变得明显。这些结果表明，该共培养系统可用于研究健康和疾病中涉及的人类神经元-OPC相互作用机制。
