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

基本信息

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

项目摘要

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

该项目的目的是开发一个3D球形核壳结构水凝胶基于水凝胶的诱导多能茎（IPS）细胞培养平台，用于受控IPS的传播和分化。我们的系统允许在水凝胶珠的中心（核）（核心）的原位生成，并通过调节藻酸盐内核成分来控制细胞外环境，以此方式以受控的和体内模拟方式以EB的分化为导向。 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 developing 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)特征…更多的Ze EB形成，细胞增殖和分化。特别是，对于第1部分，我们通过共同将氧化藻基质基质（ECM）蛋白基序（例如RGD和Yigsr Pepper）连接到藻类分子的链中，从而实现了合成的修饰的藻酸盐凝胶。我们能够成功获得具有修改的内核成分的核心壳结构水凝胶珠。在第二部分中，我们发现，通过更改内部核心组成，我们能够控制珠子内EB成员的大小。 0.5％藻酸盐核心内部的细胞形成了最大的〜600UM（直径）EBS和1％藻酸盐内的细胞，并在1：1的比率核心在1：1的藻酸盐中形成最小的〜250UM（直径）EBS。内核液化和修饰会影响EB分化的形态和轨迹。 PCR的结果表明，液化内核增强了EB分化向内皮层谱系，从1％降低到0.5％，抑制了臀部细胞分化，并以RGD辣椒派生的衍生分化向外皮毛皮谱系修饰藻酸盐。较少的