Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA; Stanford Brain Organogenesis Program, Wu Tsai Neurosciences Institute, Stanford University, Stanford, CA 94305, USA.
Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA.
Cell. 2020 Dec 23;183(7):1913-1929.e26. doi: 10.1016/j.cell.2020.11.017. Epub 2020 Dec 16.
Neurons in the cerebral cortex connect through descending pathways to hindbrain and spinal cord to activate muscle and generate movement. Although components of this pathway have been previously generated and studied in vitro, the assembly of this multi-synaptic circuit has not yet been achieved with human cells. Here, we derive organoids resembling the cerebral cortex or the hindbrain/spinal cord and assemble them with human skeletal muscle spheroids to generate 3D cortico-motor assembloids. Using rabies tracing, calcium imaging, and patch-clamp recordings, we show that corticofugal neurons project and connect with spinal spheroids, while spinal-derived motor neurons connect with muscle. Glutamate uncaging or optogenetic stimulation of cortical spheroids triggers robust contraction of 3D muscle, and assembloids are morphologically and functionally intact for up to 10 weeks post-fusion. Together, this system highlights the remarkable self-assembly capacity of 3D cultures to form functional circuits that could be used to understand development and disease.
大脑皮层中的神经元通过下行通路与后脑和脊髓相连，从而激活肌肉并产生运动。虽然该通路的组成部分以前已经在体外生成并进行了研究，但尚未用人细胞实现这个多突触回路的组装。在这里，我们生成类似于大脑皮层或后脑/脊髓的类器官，并将它们与人类骨骼肌球体组装在一起，以生成 3D 皮质运动类器官。通过狂犬病毒追踪、钙成像和膜片钳记录，我们表明皮质投射神经元投射并与脊髓球体相连，而脊髓衍生的运动神经元与肌肉相连。谷氨酸光解或皮质球体的光遗传学刺激会引发 3D 肌肉的强烈收缩，融合后，类器官的形态和功能可保持完整长达 10 周。总之，该系统突出了 3D 培养物形成功能性回路的惊人自组装能力，可用于了解发育和疾病。
