Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells
Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells
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DOI：
10.21769/bioprotoc.3350
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发表时间：
2019-09-05
期刊：
影响因子：
0.8
通讯作者：
Heine, Vivi M.
中科院分区：
文献类型：
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作者：
Dooves, Stephanie;Nadadhur, Aishwarya G.;Heine, Vivi M.
神经元和少突胶质细胞之间的相互作用对于大脑的正常运作是很重要的。已经开发了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，这些方法中的大多数包含来自动物模型的细胞。在目前的协议中，我们共同培养人类神经元与人类少突胶质细胞。神经元和少突胶质细胞前体细胞（OPC）分别从多能干细胞分化根据先前公布的协议。为了研究神经元-神经胶质串扰，将神经元和OPC在优化条件下以共培养模式铺板另外28天，并制备用于OPC成熟和神经元形态学分析。据我们所知，这是第一个包含所有人类细胞的神经元OPC协议之一。当TSC神经元与TSC OPCs共培养时，在单培养的硬化复合体（TSC）神经元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人类神经元与OPC的相互作用机制，参与健康和疾病。
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 cocultured 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.