High-throughput generation of midbrain dopaminergic neuron organoids from reporter human pluripotent stem cells.

High-throughput generation of midbrain dopaminergic neuron organoids from reporter human pluripotent stem cells.
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DOI:
10.1016/j.xpro.2021.100463
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发表时间:
2021-06-18
期刊:
影响因子:
--
通讯作者:
Ahfeldt T
Ahfeldt T
中科院分区:
其他
文献类型:
--
作者:
Sarrafha L;Parfitt GM;Reyes R;Goldman C;Coccia E;Kareva T;Ahfeldt T

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在这里,我们描述了一个高通量的3D分化方案,用于从人多能干细胞中获得中脑多巴胺能神经元。类器官的使用在疾病建模中已变得普遍,但对更同质的培养物的需求很高。我们的方法有利于大规模生产可以以不同形式维持的均匀中脑类器官,并且我们的报告子允许多巴胺能神经元的分选。成熟的长期类器官培养物可用作整个中脑的模型。有关本方案使用和执行的完整详细信息,请参阅。在转瓶中从hPSC高通量生成3D中脑类器官生成用于多巴胺能神经元的报告基因hPSC系以2D、2. 5D和3D形式培养的多巴胺能神经元在长期中脑类器官培养物中的成熟在此,我们描述了用于从人多能干细胞衍生中脑多巴胺能神经元的高通量3D分化方案。类器官的使用在疾病建模中已变得普遍,但对更同质的培养物的需求很高。我们的方法有利于大规模生产可以以不同形式维持的均匀中脑类器官,并且我们的报告子允许多巴胺能神经元的分选。成熟的长期类器官培养物可用作整个中脑的模型。
Here, we describe a high-throughput 3D differentiation protocol for deriving midbrain dopaminergic neurons from human pluripotent stem cells. The use of organoids has become prevalent in disease modeling, but there is a high demand for more homogeneous cultures. Our approach is advantageous for large-scale production of uniform midbrain organoids that can be maintained in diverse formats, and our reporters allow for sorting of dopaminergic neurons. The maturing long-term organoid cultures can be used as a model for the entire midbrain. For complete details on the use and execution of this protocol, please refer to. High-throughput generation of 3D midbrain organoids from hPSCs in spinner flasks Generation of reporter hPSC lines for dopaminergic neurons Dopaminergic neurons cultured in 2D, 2.5D, and 3D formats Maturation in long-term midbrain organoid cultures Here, we describe a high-throughput 3D differentiation protocol for deriving midbrain dopaminergic neurons from human pluripotent stem cells. The use of organoids has become prevalent in disease modeling, but there is a high demand for more homogeneous cultures. Our approach is advantageous for large-scale production of uniform midbrain organoids that can be maintained in diverse formats, and our reporters allow for sorting of dopaminergic neurons. The maturing long-term organoid cultures can be used as a model for the entire midbrain.
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