Three-dimensional neural constructs: a novel platform for neurophysiological investigation
Three-dimensional neural constructs: a novel platform for neurophysiological investigation
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DOI：
10.1088/1741-2560/5/3/006
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发表时间：
2008-09-01
影响因子：
4
通讯作者：
LaPlaca, Michelle C.
中科院分区：
文献类型：
--
作者：
Irons, Hillary R.;Cullen, D. Kacy;LaPlaca, Michelle C.
神经细胞的形态学和电生理学特性受到其直接的细胞外环境的影响，但这种环境的特征很难在体外模拟。因此，迫切需要开发新一代的培养系统，更接近模拟神经组织的复杂性。为此，我们设计了新的电生理活性的3D神经结构，由神经元和星形胶质细胞组成的生物活性细胞外基质为基础的支架。这些结构中的神经元表现出广泛的3D神经突生长，表达成熟的神经元特异性细胞骨架蛋白，并保持活力数周。此外，神经元呈现复杂的3D形态，具有丰富的神经突分支和明确的网络连接指示，包括突触连接。此外，我们修改了全细胞膜片钳技术，允许电生理探测的神经元深处的3D结构，揭示了这些神经元显示自发和诱发的电生理动作电位，并表现出功能性突触的形成和网络特性。这是第一次报告的个别膜片钳记录的神经元深处的三维支架。这些组织工程细胞构建体为神经生物学和电生理学研究提供了创新平台，为开发更具生理相关性的神经组织模型迈出了重要一步。
Morphological and electrophysiological properties of neural cells are substantially influenced by their immediate extracellular surroundings, yet the features of this environment are difficult to mimic in vitro. Therefore, there is a tremendous need to develop a new generation of culture systems that more closely model the complexity of nervous tissue. To this end, we engineered novel electrophysiologically active 3D neural constructs composed of neurons and astrocytes within a bioactive extracellular matrix-based scaffold. Neurons within these constructs exhibited extensive 3D neurite outgrowth, expressed mature neuron-specific cytoskeletal proteins, and remained viable for several weeks. Moreover, neurons assumed complex 3D morphologies with rich neurite arborization and clear indications of network connectivity, including synaptic junctures. Furthermore, we modified whole-cell patch clamp techniques to permit electrophysiological probing of neurons deep within the 3D constructs, revealing that these neurons displayed both spontaneous and evoked electrophysiological action potentials and exhibited functional synapse formation and network properties. This is the first report of individual patch clamp recordings of neurons deep within 3D scaffolds. These tissue engineered cellular constructs provide an innovative platform for neurobiological and electrophysiological investigations, serving as an important step towards the development of more physiologically relevant neural tissue models.