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的诱导多能干细胞（iPS）培养平台，用于可控的iPS增殖和分化。我们的系统允许在水凝胶珠的中心（核心）原位产生胚胎体（EB），并通过调节藻酸盐内核组成来控制细胞外环境，以这种方式以受控和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，并成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，进行了两方面的研究：1）用于iPS细胞包封的改性水凝胶基质的开发，其允许控制EB形成和分化，2）特征， ...更多信息 具体地，对于第一部分，我们已经实现了通过将牛胶原基质（ECM）蛋白基序如RGD和YIGSR肽共价连接到藻酸盐分子链来合成改性的藻酸盐凝胶。我们能够成功地得到具有改性内核组成的核壳结构水凝胶珠。对于第二部分，我们已经发现，通过改变内核组成，我们能够控制珠粒内部形成的EB的大小。在0.5%海藻酸钠芯中的细胞形成最大的~ 600 μ m（直径）EBs，在1%海藻酸钠与0.5%HA以1：1比例混合的芯中的细胞形成最小的~250 μ m（直径）EBs。内核的修饰和修饰可以影响EB分化的形态和轨迹。PCR结果表明，去除内核可促进EB向内胚层分化，降低海藻酸钠浓度（1%~ 0.5%）可抑制hiPS细胞分化，而用RGD肽修饰海藻酸钠可诱导EB向外胚层分化。少