Brain-stiffness-mimicking tilapia collagen gel promotes the induction of dorsal cortical neurons from human pluripotent stem cells
Brain-stiffness-mimicking tilapia collagen gel promotes the induction of dorsal cortical neurons from human pluripotent stem cells
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DOI：
10.1038/s41598-018-38395-5
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发表时间：
2019-02-28
影响因子：
4.6
通讯作者：
Kosodo, Yoichi
中科院分区：
文献类型：
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作者：
Iwashita, Misato;Ohta, Hatsumi;Kosodo, Yoichi
细胞外微环境的机械特性，包括其刚度，在干细胞命运决定中起着至关重要的作用。虽然先前的研究已经证明发育中的大脑在刚度方面表现出时空多样性，但仍然不清楚刚度如何调节干细胞对特定神经谱系的命运。在这里，我们建立了一个培养基板，再现了在体外重建测定使用罗非鱼胶原蛋白的脑组织的硬度。通过添加交联剂，我们获得了与活脑组织硬度相似的凝胶（150-1500 Pa）。我们进一步研究了凝胶作为干细胞培养基质的能力以及刚度对使用人iPS细胞的神经谱系分化的影响。令人惊讶的是，在神经诱导的早期暴露于硬度约为1500 Pa的凝胶促进了背皮层神经元的产生。这些发现表明，脑硬度模拟凝胶有潜力确定终端神经亚型。两者合计，交联的罗非鱼胶原蛋白凝胶预期可用于各种重建测定，可用于探索刚度在神经发生和神经功能中的作用。背皮层神经元的增强的产生也可以为神经再生应用提供相当大的优势。
The mechanical properties of the extracellular microenvironment, including its stiffness, play a crucial role in stem cell fate determination. Although previous studies have demonstrated that the developing brain exhibits spatiotemporal diversity in stiffness, it remains unclear how stiffness regulates stem cell fate towards specific neural lineages. Here, we established a culture substrate that reproduces the stiffness of brain tissue using tilapia collagen for in vitro reconstitution assays. By adding crosslinkers, we obtained gels that are similar in stiffness to living brain tissue (150-1500 Pa). We further examined the capability of the gels serving as a substrate for stem cell culture and the effect of stiffness on neural lineage differentiation using human iPS cells. Surprisingly, exposure to gels with a stiffness of approximately 1500 Pa during the early period of neural induction promoted the production of dorsal cortical neurons. These findings suggest that brain-stiffness-mimicking gel has the potential to determine the terminal neural subtype. Taken together, the crosslinked tilapia collagen gel is expected to be useful in various reconstitution assays that can be used to explore the role of stiffness in neurogenesis and neural functions. The enhanced production of dorsal cortical neurons may also provide considerable advantages for neural regenerative applications.