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DOI: 10.3791/56358-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
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.
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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