The development of 3D Cell-printing technology contributes to the application of tissue constructs in vitro in neuroscience. Collecting neural cells from patients is an efficient way of monitoring health of an individual target, which, in turn, benefits the enhancement of medicines. The fabricated sample of neural cells is exposed to potential drugs for the analysis of neuron responses. 3D cell-printing as an emerging biofabrication technology has been widely used to mimic natural 3D models in in vitro tissue research, especially in vitro brain-like tissue constructs in neuroscience. Fabricated brain-like tissue constructs provide a 3D microenvironment for primary neural cells to grow within. After more than several weeks of in vitro culturing, the formation of neural circuits in structures equips them with the capability of sensitively responding to a stimulus. In this study, an in vitro layered brain-like tissue construct is first proposed and later developed by 3D cell-printing technology. The layered structure is systematically analyzed, starting from printing parameter optimization to biological functionality performance. The optimized diameter of printing nozzle and printing speed are 0.51 mm and 5 μl s-1, respectively, and the elastic modulus is approximately 6 kPa. Live/dead and immunostaining imaging is used to verify the growth of neural cells in the printed structure. The survival rate of neural cells in 2D and 3D samples is compared, and the results demonstrate that the 3D-printed structures exhibit a better artificial culturing environment and a higher survival rate. Both 2D and 3D samples are directly cultured in a 4 × 4 multiple electrode array. Local field potentials are collected and validated by the Med64 recording system. Tetrodotoxin is used to test the drug sensitivity of the printed structure, and the excitatory postsynaptic potential signals of the physiological performance indicate that the 3D-printed structure has great potential as a drug testing model in the pharmaypeceutical study.

中文翻译：

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通过3D细胞打印技术对类似大脑的组织构造进行工程设计。

3D细胞打印技术的发展有助于组织构造物在神经科学中的体外应用。从患者那里收集神经细胞是监测单个靶标健康的有效方法，从而有利于增强药物。将制造的神经细胞样本暴露于潜在药物以分析神经元反应。3D细胞打印作为一种新兴的生物制造技术已被广泛用于模拟体外组织研究中的自然3D模型，尤其是神经科学中的体外脑样组织构造。预制的大脑样组织构造可为原代神经细胞在其中生长提供3D微环境。经过数周的体外培养，结构中神经回路的形成使它们具有对刺激敏感地反应的能力。在这项研究中，首先提出了体外分层的脑样组织构造，然后通过3D细胞打印技术进行了开发。从打印参数优化到生物学功能性能，系统地分析了分层结构。最佳的打印喷嘴直径和打印速度分别为0.51 mm和5μls-1，弹性模量约为6 kPa。活/死和免疫染色成像用于验证印刷结构中神经细胞的生长。比较了2D和3D样本中神经细胞的存活率，结果表明3D打印的结构表现出更好的人工培养环境和更高的存活率。2D和3D样品都直接培养在4×4多电极阵列中。Med64记录系统收集并验证了局部场电势。河豚毒素被用于测试印刷结构的药物敏感性，并且具有生理性能的兴奋性突触后电位信号表明3D打印的结构作为药典研究中的药物测试模型具有巨大的潜力。