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

该项目的目标是开发基于水凝胶的 3D 球形核壳结构诱导多能干 (iPS) 细胞培养平台，用于受控 iPS 增殖和分化。我们的系统允许在水凝胶珠的中心（核心）原位生成胚体（EB），并通过调整藻酸盐内核成分来控制细胞外环境，从而以受控和体内模拟的方式指导EB的分化。该系统预计将成为 iPS 细胞分化的更有效、高效和仿生平台，这可能最终使再生医学研究界受益，并作为发育生物学家更好地了解胚胎发育的宝贵工具。在 2013 财年，进行了两项研究：1）开发用于 iPS 细胞封装的改良水凝胶基质，从而控制 EB 形成和分化， 2) 表征 EB 形成、细胞增殖和分化。具体来说，在第 1 部分中，我们通过将 RGD 和 YIGSR 肽等胞外基质 (ECM) 蛋白基序共价连接到藻酸盐分子链上，实现了改性藻酸盐凝胶的合成。我们能够成功地获得具有改性内核成分的核壳结构水凝胶珠。对于第二部分，我们发现通过改变内核成分，我们能够控制珠子内部形成的 EB 的大小。 0.5%藻酸盐核心内的细胞形成最大~600um（直径）的EB，而1%藻酸盐与0.5%HA以1:1比例混合的核心内的细胞形成最小~250um（直径）EB。内核的液化和改性可以影响EB分化的形态和轨迹。 PCR结果表明，液化内核增强了EB向内胚层谱系的分化，将藻酸盐浓度从1%降低至0.5%抑制了hiPS细胞的分化，并且用RGD肽修饰藻酸盐诱导向外胚层谱系的分化。较少的