Viscoelasticity and Adhesion Signaling in Biomaterials Control Human Pluripotent Stem Cell Morphogenesis in 3D Culture
Viscoelasticity and Adhesion Signaling in Biomaterials Control Human Pluripotent Stem Cell Morphogenesis in 3D Culture
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DOI：
10.1002/adma.202101966
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发表时间：
2021-09-09
影响因子：
29.4
通讯作者：
Chaudhuri, Ovijit
中科院分区：
文献类型：
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作者：
Indana, Dhiraj;Agarwal, Pranay;Chaudhuri, Ovijit
类器官是包含管腔的多细胞结构，概括了器官的关键特征，并且越来越多地用于疾病模型，药物测试和再生医学。最近的工作已经使用3D培养模型在重构基底膜（rBM）基质中从人诱导多能干细胞（hiPSC）形成类器官。然而，rBM矩阵对微环境的控制很少。更一般地，基质粘弹性在引导管腔形成中的作用仍然未知。在这里，具有独立可调的应力松弛（粘弹性）、刚度和精氨酸-甘氨酸-天冬氨酸（RGD）配体密度的粘弹性藻酸盐水凝胶用于研究3D培养中的hiPSC形态发生。一个相图，显示这些属性如何控制hiPSC形态发生的报告。较高的RGD密度和快速应力松弛促进hiPSC活力、增殖、顶基底极化和管腔形成，而低RGD密度下的缓慢应力松弛触发hiPSC凋亡。值得注意的是，hiPSC在藻酸盐水凝胶中保持多能性的时间比在rBM基质中报道的时间长得多。管腔形成受肌动球蛋白收缩性调节，并伴随着Yes相关蛋白（雅普）从细胞核易位到细胞质。结果揭示了基质粘弹性是调节干细胞形态发生的一个有效因素，并为如何利用工程生物材料构建类器官提供了新的见解。
Organoids are lumen-containing multicellular structures that recapitulate key features of the organs, and are increasingly used in models of disease, drug testing, and regenerative medicine. Recent work has used 3D culture models to form organoids from human induced pluripotent stem cells (hiPSCs) in reconstituted basement membrane (rBM) matrices. However, rBM matrices offer little control over the microenvironment. More generally, the role of matrix viscoelasticity in directing lumen formation remains unknown. Here, viscoelastic alginate hydrogels with independently tunable stress relaxation (viscoelasticity), stiffness, and arginine-glycine-aspartate (RGD) ligand density are used to study hiPSC morphogenesis in 3D culture. A phase diagram that shows how these properties control hiPSC morphogenesis is reported. Higher RGD density and fast stress relaxation promote hiPSC viability, proliferation, apicobasal polarization, and lumen formation, while slow stress relaxation at low RGD densities triggers hiPSC apoptosis. Notably, hiPSCs maintain pluripotency in alginate hydrogels for much longer times than is reported in rBM matrices. Lumen formation is regulated by actomyosin contractility and is accompanied by translocation of Yes-associated protein (YAP) from the nucleus to the cytoplasm. The results reveal matrix viscoelasticity as a potent factor regulating stem cell morphogenesis and provide new insights into how engineered biomaterials may be leveraged to build organoids.