光駆動型分化に向けた金ナノ粒子含有ヒドロゲルへの胚様体包接

含金纳米颗粒水凝胶中的胚状包涵体用于光驱动分化

• 批准号: 12F02388
• 负责人: 中辻 憲夫
• 金额: $ 1.02万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12F02388/
• 关键词: iPS cells; Controlled differentiation; Algniate 2D matrix; 人工多能性幹(iPS)細胞; 金ナノロッド(GNR); 分化制御; 3次元培養

The goal of this project is to develop a 3D spherical core-shell structure hydrogel-based induced pluripotent stem (iPS) cell culture platform for controlled iPS propagation and diflerentiation. Our system allows the in situ generation of embryo body (EB) at the center (core) of hydrogel beads and the control of extracellular environment by adjusting alginate inner core composition, in such way todirect the differentiation of EB in a controlled, and in-vivo mimic manner. This proposed system is expected to become a more effective, efficient, and biomimetic platform for iPS cell differentiation, which might ultimately benefit the regenerative medicine research community, and serve as a valuabletool for developmental biologist for better understanding of the development of embryo.ln 2013 fiscal year, two folds of studies have been carried out : 1) development of modified hydrogel matrixes for iPS cells encapsulation, which allow controlling EB formation and differentiation, 2) characteri … More ze EB formation, cell Proliferation and differentiation.Specifically, for part 1, we have achieved synthesizing modified alginate gel by covalently attaching oxtracollular matrix (ECM) protein motifs such as RGD and YIGSR peptide to the chain of alginate molecules. We were able to successfully got core shell structure hydrogel beads with modified inner core composition. For the second part, we have found that by changing the inner core composition, we were able to control the size of EB fomed inside our beads. Cells inside 0.5% alginate core formed the biggest~600um (diameter) EBs and cells inside 1% alginate mixed with 0.5%HA at 1:1 ratio core formed smallest~250 um (diameter) EBs. The inner core liquefying and modification can affect EB differentiation morphology and trajectory. PCR results suggested that liquefying inner core enhanced EB differentiation towards endodermal lineage, decreasing alginate concentration from 1 percent to 0.5 peroent suppressed hiPS cell differentiation, and modifying alginate with RGD peptide induced differentiation towards ectodermal lineage. Less

本项目的目标是开发一种基于水凝胶的三维球形核壳结构诱导多能干细胞培养平台，用于可控的诱导多能干细胞的增殖和分化。我们的系统允许在水凝胶微珠的中心(核心)原位产生胚体(EB)，并通过调整藻酸盐核心组成来控制细胞外环境，以这种方式以受控的、体内模拟的方式指导EB的分化。该系统有望成为一个更有效、高效和仿生的iPS细胞分化平台，最终可能使再生医学研究界受益，并为发育生物学家更好地了解胚胎的发育提供一个有价值的工具。2013财年，开展了两项研究：1)开发用于iPS细胞包埋的改良水凝胶基质，允许控制EB的形成和分化；2)特征…具体地说，在第一部分，我们通过将RGD和YIGSR等蛋白质基序共价连接到海藻酸分子链上，合成了改性的海藻酸凝胶。我们成功地得到了具有核壳结构的水凝胶微珠，并对其内核组成进行了改性。对于第二部分，我们发现通过改变内核组成，我们能够控制在我们的珠子中形成的EB的大小。0.5%海藻酸盐芯中的细胞形成最大的~600um(直径)EBS，1%海藻酸盐与0.5%HA按1：1比例混合形成最小的~250um(直径)EBS。内芯液化和改性可以影响EB的分化形态和轨迹。聚合酶链式反应结果表明，液化内核能促进EB向内胚层分化，海藻酸盐浓度从1%降至0.5%抑制HIPS细胞分化，RGD多肽修饰海藻酸诱导向外胚层分化。较少