Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells.
Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells.
复制标题
DOI：
10.1038/nprot.2018.006
复制
发表时间：
2018-04
期刊：
影响因子：
14.8
通讯作者：
Ma Z
中科院分区：
文献类型：
--
作者：
Hoang P;Wang J;Conklin BR;Healy KE;Ma Z
人类诱导多能干细胞(HiPSCs)的发现为研究组织形态发生和器官发育提供了前所未有的机会。目前心脏器官工程的方法要么依赖于胚胎样体的直接心脏分化，要么依赖于从单层hPSCs的预分化心肌细胞产生排列好的心脏组织。为了在体外对早期心脏器官发生进行实验建模，我们的方案结合了基于生物材料的细胞模式和干细胞器官工程。三维心脏微腔是由二维的HiPSC克隆形成的，它与早期发育的心脏相似，具有明显的空间组织和自组装。经过适当的光刻微制造、人类多能干细胞的维持和心脏分化方面的培训，一名有指导的研究生很可能能够实施这一实验方案，这需要大约三周的时间。我们预计，这种人类早期心脏发育的体外模型可以作为胚胎毒性的筛选试验，用于药物发现、调控和处方以促进胎儿的健康发育。目前，该方法还没有被用于评估遗传性疾病的HiPSC系心脏微腔的形成，这可能被用于研究胚胎发育早期心脏畸形的疾病机制。该方案描述了从人类多能干细胞产生的早期发育的心脏器官。HPSCs的几何限制将细胞的空间组织从2D层驱动到3D心脏微腔。
The discovery of human induced pluripotent stem cells (hiPSCs) has provided an unprecedented opportunity to study tissue morphogenesis and organ development as “organogenesis-in-a-dish”. Current approaches of cardiac organoid engineering rely on either direct cardiac differentiation from embryoid bodies or generation of aligned cardiac tissues from pre-differentiated cardiomyocytes from monolayer hiPSCs. To experimentally model early cardiac organogenesis in vitro, our protocol combines biomaterials-based cell patterning with stem cell organoid engineering. Three-dimensional cardiac microchambers are created from two-dimensional hiPSC colonies, which approximates an early developing heart with distinct spatial organization and self-assembly. With proper training on photolithography microfabrication, maintenance of human pluripotent stem cells, and cardiac differentiation, a graduate student with guidance will likely be able to carry out this experimental protocol, which requires approximately three weeks. We envisage that this in vitro model of human early heart development could serve as an embryotoxicity screening assay in drug discovery, regulation, and prescription for healthy fetal development. Currently, this protocol has yet to be implemented to assess the cardiac microchamber formation from the hiPSC lines with inherited disease, which can potentially be used to study the disease mechanisms of cardiac malformations at early stage of embryogenesis. This protocol describes the generation early developing cardiac organoids from human pluripotent stem cells. Geometric confinement of the hiPSCs drives spatial organization of the cells from a 2D layer into 3D cardiac microchambers.
登录
查看更多内容
影响因子：
14
作者：
Ma, Zhen;Koo, Sangmo;Finnegan, Micaela A.;Loskill, Peter;Huebsch, Nathaniel;Marks, Natalie C.;Conklin, Bruce R.;Grigoropoulos, Costas P.;Healy, Kevin E.
通讯作者：
Healy, Kevin E.
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.6
作者：
Lee, Eugene K.;Kurokawa, Yosuke K.;Khine, Michelle
通讯作者：
Khine, Michelle
影响因子：
48
作者：
Burridge, Paul W.;Matsa, Elena;Shukla, Praveen;Lin, Ziliang C.;Churko, Jared M.;Ebert, Antje D.;Lan, Feng;Diecke, Sebastian;Huber, Bruno;Mordwinkin, Nicholas M.;Plews, Jordan R.;Abilez, Oscar J.;Cui, Bianxiao;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
41.2
作者：
Caiazzo, Massimiliano;Okawa, Yuya;Lutolf, Matthias P.
通讯作者：
Lutolf, Matthias P.