Nagashima Fumiaki, Suzuki Ikuo K, Shitamukai Atsunori, Sakaguchi Haruko, Iwashita Misato, Kobayashi Taeko, Tone Shigenobu, Toida Kazunori, Vanderhaeghen Pierre, Kosodo Yoichi
1 Department of Anatomy, Kawasaki Medical School , Kurashiki, Japan .
Stem Cells Dev. 2014 Sep 15;23(18):2129-42. doi: 10.1089/scd.2013.0251. Epub 2014 Jan 31.
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.
当前的干细胞技术已能够在体外从胚胎干细胞(ESC)和诱导多能干细胞诱导生成皮质祖细胞和神经元。为了理解在单层培养中生成的皮质细胞干性相关的顶-基极性获得及突起形成的潜在机制,在此,我们基于神经上皮组织中黏附连接的适度解离开发了一种新型的子宫内移植系统。该方法能够实现:(1)显著增加移植细胞的掺入数量;(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,我们将从小鼠ESC诱导生成的皮质祖细胞移植到发育中的大脑。重要的是,我们发现突起延伸模式取决于宿主上皮组织的外在顶-基极性以及移植细胞的内在分化状态。此外,我们成功移植了从人ESC诱导生成的皮质祖细胞,表明我们的策略能够研究发育中的小鼠皮质内人神经祖细胞的神经发生。具体而言,人皮质细胞表现出多种放射状迁移特征。在此建立的强大移植方法既可以用于揭示ESC神经发生与组织环境之间缺失的差距,也可以作为正常和病理性人皮质发生的体内模型。