Graduate Program in Neuroscience and Behavior, UMass Amherst, Amherst, MA 01003, USA; Department of Biochemistry and Molecular Biology, UMass Amherst, Amherst, MA 01003, USA.
Department of Biochemistry and Molecular Biology, UMass Amherst, Amherst, MA 01003, USA.
Cell Rep Methods. 2024 Jan 22;4(1):100689. doi: 10.1016/j.crmeth.2023.100689. Epub 2024 Jan 15.
In early neurodevelopment, the central nervous system is established through the coordination of various neural organizers directing tissue patterning and cell differentiation. Better recapitulation of morphogen gradient production and signaling will be crucial for establishing improved developmental models of the brain in vitro. Here, we developed a method by assembling polydimethylsiloxane devices capable of generating a sustained chemical gradient to produce patterned brain organoids, which we termed morphogen-gradient-induced brain organoids (MIBOs). At 3.5 weeks, MIBOs replicated dorsal-ventral patterning observed in the ganglionic eminences (GE). Analysis of mature MIBOs through single-cell RNA sequencing revealed distinct dorsal GE-derived CALB2 interneurons, medium spiny neurons, and medial GE-derived cell types. Finally, we demonstrate long-term culturing capabilities with MIBOs maintaining stable neural activity in cultures grown up to 5.5 months. MIBOs demonstrate a versatile approach for generating spatially patterned brain organoids for embryonic development and disease modeling.
在早期神经发育过程中，中枢神经系统通过各种神经组织者的协调来建立，这些组织者指导组织模式形成和细胞分化。更好地再现形态发生梯度的产生和信号传递对于建立体外大脑发育模型至关重要。在这里，我们开发了一种通过组装聚二甲基硅氧烷器件来产生持续化学梯度的方法，从而产生图案化的脑类器官，我们将其称为形态发生梯度诱导的脑类器官（MIBO）。在 3.5 周时，MIBO 复制了神经节隆起（GE）中观察到的背腹模式。通过单细胞 RNA 测序对成熟 MIBO 的分析显示出明显的背侧 GE 衍生的 CALB2 中间神经元、中等棘突神经元和内侧 GE 衍生的细胞类型。最后，我们证明了 MIBO 具有长期培养能力，在培养物中培养长达 5.5 个月，其神经活动保持稳定。MIBO 为胚胎发育和疾病建模生成空间图案化脑类器官提供了一种通用方法。
