Large-Scale Production of Mature Neurons from Human Pluripotent Stem Cells in a Three-Dimensional Suspension Culture System.
Large-Scale Production of Mature Neurons from Human Pluripotent Stem Cells in a Three-Dimensional Suspension Culture System.
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DOI：
10.1016/j.stemcr.2016.05.010
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发表时间：
2016-06-14
影响因子：
5.9
通讯作者：
Rubin LL
中科院分区：
文献类型：
--
作者：
Rigamonti A;Repetti GG;Sun C;Price FD;Reny DC;Rapino F;Weisinger K;Benkler C;Peterson QP;Davidow LS;Hansson EM;Rubin LL
人类多能干细胞（hPSC）提供了一种可再生的细胞来源，可以无限扩增并分化成人体内几乎任何类型的细胞，包括神经元。这为研究神经元细胞和发育生物学以及神经系统的细胞病理学开辟了前所未有的可能性，为筛选影响这些过程的化学文库提供了平台，并为神经系统疾病的再生方法提供了可移植细胞的潜在来源。然而，定义允许大量和高产量的神经元的协议已被证明是困难的。我们目前的分化协议的不同亚型的神经元的生成在一个高度可重复的方式，与最小的实验到实验的变化。这些神经元与邻近的细胞形成突触，表现出自发的电活动，并对去极化作出适当的反应。hPSC衍生的神经元表现出高度成熟，并且即使没有星形胶质细胞饲养层，也能在培养物中存活长达4-5个月。hPSC高效、大规模分化为不同的神经元亚型分化的神经元可以在转瓶中维持较长的时间衍生的神经元功能成熟并形成广泛的突触网络用于不同形式的神经系统疾病体外建模的有吸引力的平台Ruben和同事提出了从人类多能干细胞大规模生产皮质和运动神经元的新方案。所获得的神经元是功能性的，在不使用饲养层的情况下在培养中存活延长的时间段，并且构成了用于神经系统疾病的体外建模的有希望的平台。
Human pluripotent stem cells (hPSCs) offer a renewable source of cells that can be expanded indefinitely and differentiated into virtually any type of cell in the human body, including neurons. This opens up unprecedented possibilities to study neuronal cell and developmental biology and cellular pathology of the nervous system, provides a platform for the screening of chemical libraries that affect these processes, and offers a potential source of transplantable cells for regenerative approaches to neurological disease. However, defining protocols that permit a large number and high yield of neurons has proved difficult. We present differentiation protocols for the generation of distinct subtypes of neurons in a highly reproducible manner, with minimal experiment-to-experiment variation. These neurons form synapses with neighboring cells, exhibit spontaneous electrical activity, and respond appropriately to depolarization. hPSC-derived neurons exhibit a high degree of maturation and survive in culture for up to 4–5 months, even without astrocyte feeder layers. Efficient, large-scale differentiation of hPSC to distinct neuronal subtypes Differentiated neurons can be maintained in spinner flasks for extended time periods Derived neurons are functionally mature and form an extensive synaptic network Attractive platform for in vitro modeling of different forms of neurological disease Ruben and colleagues present new protocols for large-scale production of cortical and motor neurons from human pluripotent stem cells. The obtained neurons are functional, survive an extended period of time in culture without the use of feeder layers, and constitute a promising platform for in vitro modeling of neurological diseases.