Novel and Robust Transplantation Reveals the Acquisition of Polarized Processes by Cortical Cells Derived from Mouse and Human Pluripotent Stem Cells
Novel and Robust Transplantation Reveals the Acquisition of Polarized Processes by Cortical Cells Derived from Mouse and Human Pluripotent Stem Cells
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DOI：
10.1089/scd.2013.0251
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发表时间：
2014-09-15
影响因子：
4
通讯作者：
Kosodo, Yoichi
中科院分区：
文献类型：
--
作者：
Nagashima, Fumiaki;Suzuki, Ikuo K.;Kosodo, Yoichi
目前的干细胞技术已经能够从胚胎干细胞（ESC）和体外诱导多能干细胞诱导皮质祖细胞和神经元。要了解的机制，收购apico-basal极性和形成的过程中产生的皮层细胞的干细胞单层培养，在这里，我们开发了一种新的子宫内移植系统的基础上适度解离的粘附连接在神经上皮组织。该方法能够（1）掺入显著更高数量的移植细胞和（2）以单细胞分辨率进行定量形态学分析，包括延时记录分析。然后，我们将小鼠胚胎干细胞诱导的皮质祖细胞移植到发育中的大脑中。重要的是，我们发现，过程扩展的模式取决于宿主上皮组织的外在apico-basal极性，以及移植细胞的内在分化状态。此外，我们成功地移植了从人类胚胎干细胞诱导的皮质祖细胞，表明我们的策略能够研究发育中的小鼠皮质内人类神经祖细胞的神经发生。具体而言，人类皮质细胞表现出放射状迁移的多种特征。这里建立的强大的移植方法可以用来揭示神经发生从胚胎干细胞和组织环境之间的缺失的差距，并作为正常和病理性人类皮质生成的体内模型。
Current stem cell technologies have enabled the induction of cortical progenitors and neurons from embryonic stem cells (ESCs) and induced pluripotent stem cells in vitro. To understand the mechanisms underlying the acquisition of apico-basal polarity and the formation of processes associated with the stemness of cortical cells generated in monolayer culture, here, we developed a novel in utero transplantation system based on the moderate dissociation of adherens junctions in neuroepithelial tissue. This method enables (1) the incorporation of remarkably higher numbers of grafted cells and (2) quantitative morphological analyses at single-cell resolution, including time-lapse recording analyses. We then grafted cortical progenitors induced from mouse ESCs into the developing brain. Importantly, we revealed that the mode of process extension depends on the extrinsic apico-basal polarity of the host epithelial tissue, as well as on the intrinsic differentiation state of the grafted cells. Further, we successfully transplanted cortical progenitors induced from human ESCs, showing that our strategy enables investigation of the neurogenesis of human neural progenitors within the developing mouse cortex. Specifically, human cortical cells exhibit multiple features of radial migration. The robust transplantation method established here could be utilized both to uncover the missing gap between neurogenesis from ESCs and the tissue environment and as an in vivo model of normal and pathological human corticogenesis.