Koch Lothar, Deiwick Andrea, Soriano Jordi, Chichkov Boris
Institut für Quantenoptik, Leibniz Universität Hannover, 30167 Hannover, Germany.
NIFE-Niedersächsisches Zentrum für Biomedizintechnik, Implantatforschung und Entwicklung, 30625 Hannover, Germany.
Int J Bioprint. 2023 Jan 18;9(2):672. doi: 10.18063/ijb.v9i2.672. eCollection 2023.
Generation of human neuronal networks by three-dimensional (3D) bioprinting is promising for drug testing and hopefully will allow for the understanding of cellular mechanisms in brain tissue. The application of neural cells derived from human induced-pluripotent stem cells (hiPSCs) is an obvious choice, since hiPSCs provide access to cells unlimited in number and cell types that could be generated by differentiation. The questions in this regard include which neuronal differentiation stage is optimal for printing of such networks, and to what extent the addition of other cell types, especially astrocytes, supports network formation. These aspects are the focus of the present study, in which we applied a laser-based bioprinting technique and compared hiPSC-derived neural stem cells (NSCs) with neuronal differentiated NSCs, with and without the inclusion of co-printed astrocytes. In this study, we investigated in detail the effects of cell types, printed droplet size, and duration of differentiation before and after printing on viability, as well as proliferation, stemness, differentiation potential, formation of dendritic extensions and synapses, and functionality of the generated neuronal networks. We found a significant dependence of cell viability after dissociation on differentiation stage, but no impact of the printing process. Moreover, we observed a dependence of the abundance of neuronal dendrites on droplet size, a marked difference between printed cells and normal cell culture in terms of further differentiation of the cells, especially differentiation into astrocytes, as well as neuronal network formation and activity. Notably, there was a clear effect of admixed astrocytes on NSCs but not on neurons.
通过三维（3D）生物打印生成人类神经元网络对于药物测试很有前景，并且有望促进对脑组织细胞机制的理解。应用源自人类诱导多能干细胞（hiPSC）的神经细胞是一个显而易见的选择，因为hiPSC提供了获取数量无限且可通过分化产生多种细胞类型的细胞的途径。这方面的问题包括哪个神经元分化阶段最适合打印此类网络，以及添加其他细胞类型，特别是星形胶质细胞，在多大程度上支持网络形成。这些方面是本研究的重点，在本研究中，我们应用了基于激光的生物打印技术，并比较了hiPSC衍生的神经干细胞（NSC）与神经元分化的NSC，以及是否共打印星形胶质细胞的情况。在这项研究中，我们详细研究了细胞类型、打印液滴大小以及打印前后的分化持续时间对活力、增殖、干性、分化潜能、树突延伸和突触形成以及所生成神经元网络功能的影响。我们发现解离后细胞活力显著依赖于分化阶段，但打印过程没有影响。此外，我们观察到神经元树突的丰度依赖于液滴大小，打印细胞与正常细胞培养在细胞进一步分化，特别是分化为星形胶质细胞方面存在显著差异，以及在神经元网络形成和活性方面也存在差异。值得注意的是，共混的星形胶质细胞对NSC有明显影响，但对神经元没有影响。
