William G. Lowrie Dept. of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA.
Biotechnol Prog. 2013 Jul-Aug;29(4):1013-22. doi: 10.1002/btpr.1742. Epub 2013 May 9.
The clinical use of pluripotent stem cell (PSC)-derived neural cells requires an efficient differentiation process for mass production in a bioreactor. Toward this goal, neural differentiation of murine embryonic stem cells (ESCs) in three-dimensional (3D) polyethylene terephthalate microfibrous matrices was investigated in this study. To streamline the process and provide a platform for process integration, the neural differentiation of ESCs was induced with astrocyte-conditioned medium without the formation of embryoid bodies, starting from undifferentiated ESC aggregates expanded in a suspension bioreactor. The 3D neural differentiation was able to generate a complex neural network in the matrices. When compared to 2D differentiation, 3D differentiation in microfibrous matrices resulted in a higher percentage of nestin-positive cells (68% vs. 54%) and upregulated gene expressions of nestin, Nurr1, and tyrosine hydroxylase. High purity of neural differentiation in 3D microfibrous matrix was also demonstrated in a spinner bioreactor with 74% nestin + cells. This study demonstrated the feasibility of a scalable process based on 3D differentiation in microfibrous matrices for the production of ESC-derived neural cells.
胚胎干细胞（PSC）衍生的神经细胞的临床应用需要在生物反应器中进行大规模生产的高效分化过程。为了实现这一目标，本研究探讨了在三维（3D）聚对苯二甲酸乙二醇酯微纤维基质中进行鼠胚胎干细胞（ESCs）的神经分化。为了简化该过程并为工艺集成提供平台，本研究从在悬浮生物反应器中扩增的未分化 ESC 聚集体开始，使用星形胶质细胞条件培养基而非胚状体形成来诱导 ESC 的神经分化。3D 神经分化能够在基质中生成复杂的神经网络。与 2D 分化相比，微纤维基质中的 3D 分化导致巢蛋白阳性细胞的比例更高（68%对 54%），并且巢蛋白、Nurr1 和酪氨酸羟化酶的基因表达上调。在搅拌生物反应器中，也证明了 3D 微纤维基质中神经分化的高纯度，其巢蛋白+细胞的比例为 74%。本研究证明了基于微纤维基质中 3D 分化的可扩展工艺生产 ESC 衍生的神经细胞的可行性。
