Novel Methods to Apply Micro Dynamic Stimulations on Cultured Adhesive Cells and Its Application in Constructing Gel-embedded Three-dimensional Neuronal Structures Differentiated From Human iPS Cells
Novel Methods to Apply Micro Dynamic Stimulations on Cultured Adhesive Cells and Its Application in Constructing Gel-embedded Three-dimensional Neuronal Structures Differentiated From Human iPS Cells
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对培养的粘附细胞进行微动态刺激的新方法及其在构建人 iPS 细胞分化的凝胶嵌入三维神经元结构中的应用
DOI：
10.1007/s00542-017-3399-4
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Kaoru Goto
中科院分区：
文献类型：
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作者：
Tadashi Kosawada;Keita Ohnishi;Hiroaki Satoh;Zhonggang Feng;Kaoru Goto
在细胞工程中，非常需要对培养的粘附细胞或细胞集落施加主动动态刺激的装置系统，以便在临床治疗（例如再生医学中的细胞移植）之前控制细胞功能以及形态。本研究开发了一种简单有效的三维微振动台和二维微振动台，用于细胞培养箱内有限的空间和环境，直接刺激培养的贴壁iPS细胞集落，促进其向目标细胞分化。开发过程中采用了有限元法。此外，结合动态刺激，提出了一种神经元凝胶包埋培养的方法来构建三维神经元结构。通过测定有无动态刺激诱导的人iPS细胞的β Ⅲ微管蛋白免疫染色面积，研究动态刺激对iPS细胞向神经细胞分化和生长过程的影响。结果发现，静态培养和动态刺激的人iPS细胞向神经元的分化和生长效率存在显著差异。此外，它被证实，凝胶包埋培养结合动态刺激是一个有前途的策略，以构建三维神经元结构。
In cellular engineering, device system to apply active dynamic stimulations on cultured adhesive cells or cell colonies is highly needed in order to control cellular functions as well as morphologies prior to clinical treatment such as cell transplantation in regenerative medicine. In this study, simple but effective cantilever-type three-dimensional micro vibration table as well as two-dimensional one to be used within limited space and environment inside the cell incubator is newly developed to stimulate cultured adhesive iPS cells colony directly to promote their differentiation into objective cells. The finite element method is utilized in developing process. Also, together with the dynamic stimulation, a method of gel-embedded culture for neurons is proposed to construct three-dimensional neuronal structures. Effects of the dynamic stimulations on the differentiation and growth process of the human iPS cells into neuronal cells are investigated by measuring the immunostained areas of βIII-tubulin of the differentiated neural cells with or without the dynamic stimulations. It is found that there is significant difference in differentiation and growth efficiency of the human iPS cells into neurons between the statically cultured cells and the dynamically stimulated ones. Furthermore, it is confirmed that the gel-embedded culture together with the dynamic stimulations is a promising strategy to construct the three-dimensional neuronal structures.