Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA.
Departments of Neurology and Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
Cell Rep Methods. 2022 Jul 18;2(7):100255. doi: 10.1016/j.crmeth.2022.100255.
Neuronal cultures provide a basis for reductionist insights that rely on molecular and pharmacological manipulation. However, the inability to culture mature CNS neurons limits our understanding of adult neuronal physiology. Here, we report methods for culturing central nervous system neurons in large numbers and across multiple brain regions for extended time periods. Primary adult neuronal cultures develop polarity; they establish segregated dendritic and axonal compartments, maintain resting membrane potentials, exhibit spontaneous and evoked electrical activity, and form neural networks. Cultured adult neurons isolated from different brain regions such as the hippocampus, cortex, brainstem, and cerebellum exhibit distinct cell morphologies, growth patterns, and spontaneous firing characteristics reflective of their regions of origin. Using adult motor cortex cultures, we identify a CNS "conditioning" effect after spinal cord injury. The ability to culture adult neurons offers a valuable tool for studying basic and therapeutic science of the brain.
神经元培养为依赖于分子和药理学操作的简化观点提供了基础。然而,培养成熟中枢神经系统神经元的能力限制了我们对成年神经元生理学的理解。在这里,我们报告了在多个脑区大量培养中枢神经系统神经元并延长培养时间的方法。原代成年神经元培养物具有极性;它们建立了分离的树突和轴突区室,保持静息膜电位,表现出自发和诱发的电活动,并形成神经网络。从不同脑区(如海马体、皮层、脑干和小脑)分离的培养成年神经元表现出独特的细胞形态、生长模式和自发放电特征,反映了它们的起源区域。使用成年运动皮层培养物,我们在脊髓损伤后鉴定出中枢神经系统“调节”效应。培养成年神经元的能力为研究大脑的基础科学和治疗科学提供了有价值的工具。