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Neuroscience
小鼠胚胎干细胞向皮质 Interneuron 前体细胞的分化
小鼠胚胎干细胞向皮质 Interneuron 前体细胞的分化
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Differentiation of Mouse Embryonic Stem Cells into Cortical Interneuron Precursors

小鼠胚胎干细胞向皮质 Interneuron 前体细胞的分化

Full Text
10,981 Views
10:24 min
December 3, 2017

DOI: 10.3791/56358-v

David J. Tischfield1,2, Stewart A. Anderson1

1Neuroscience Graduate Group, Perelman School of Medicine,University of Pennsylvania, 2Department of Psychiatry, Children's Hospital of Philadelphia,University of Pennsylvania School of Medicine

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol describes a method for generating cortical interneuron progenitors and post-mitotic interneuron precursors from mouse embryonic stem cells using a modified embryoid body-to-monolayer method. This technique enables efficient generation of Nkx2.1 expressing interneuron progenitors for studying fate determination and therapeutic applications.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Stem Cell Research

Background

  • Cortical interneurons play a crucial role in brain function.
  • Understanding their development can provide insights into neurological disorders.
  • This method allows for the study of interneuron fate determination.
  • It can also facilitate the enrichment of specific interneuron subtypes.

Purpose of Study

  • To generate cortical interneuron progenitors from mouse embryonic stem cells.
  • To investigate the developmental pathways of cortical interneurons.
  • To explore potential therapeutic applications of these progenitors.

Methods Used

  • Modified embryoid body-to-monolayer method.
  • Use of mitotically inactive MEF feeder cells.
  • Cell culture techniques at 37 degrees Celsius.
  • Fluorescent sorting of interneuron precursors.

Main Results

  • Successful generation of cortical interneuron progenitors.
  • Efficient enrichment of parvalbumin and somatostatin interneuron subgroups.
  • Insights into the fate determination of cortical interneurons.
  • Potential for therapeutic applications in neuroscience.

Conclusions

  • This method provides a reliable approach for generating interneuron progenitors.
  • It enhances our understanding of cortical interneuron development.
  • Future studies can leverage these findings for therapeutic advancements.

Frequently Asked Questions

What are cortical interneurons?
Cortical interneurons are a type of neuron that regulates the activity of other neurons in the cerebral cortex.
How does the modified embryoid body-to-monolayer method work?
This method involves transitioning embryoid bodies into a monolayer culture to promote the differentiation of stem cells into specific neuron types.
What is the significance of Nkx2.1 expressing interneuron progenitors?
Nkx2.1 expressing interneuron progenitors are important for understanding the development of specific interneuron subtypes that are crucial for brain function.
Can these progenitors be used for therapeutic applications?
Yes, the generated progenitors can potentially be used in cell replacement therapies for neurological disorders.
What are the advantages of using fluorescent sorting?
Fluorescent sorting allows for the isolation of specific cell types, enhancing the purity and efficacy of the progenitors for research and therapeutic use.

本议定书描述了一种方法, 产生皮层 interneuron 祖细胞和后有丝分裂 interneuron 前体从小鼠胚胎干细胞使用改良的胚体-单层法。这些祖/前体可用于体外或荧光分类和移植到新生儿大脑皮层研究命运确定, 或用于治疗应用。

该程序的总体目标是使用改良的胚状体到单层方法,在小鼠胚胎干细胞的有丝分裂后中间神经元前体中产生皮质中间神经元祖细胞。这种方法可以帮助回答神经科学领域的关键问题,例如皮层中间神经元的亚型如何在发育过程中实现其各自的命运。该技术的主要优点是它能够有效产生表达 Nkx2.1 的中间神经元祖细胞,以及富集小清蛋白或生长抑素中间神经元亚群的方法。

要开始此过程,将 MEF 培养基中有丝分裂失活的 MEF 饲养层细胞接种到经组织培养处理的平板上。在铺板 mESC 之前,请至少等待 12 小时,让它们在 37 摄氏度的细胞培养箱中沉淀。如果未立即使用,请每三天更换一次 MEF 介质。

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