EAGER: Mechanics-Guided Multicellular Self-Organization

EAGER：力学引导的多细胞自组织

基本信息

批准号：
1933061
负责人：
Jianping Fu
金额：
$ 29.97万
依托单位：
Regents of the University of Michigan - Ann Arbor
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-15 至 2021-12-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1933061&HistoricalAwards=false
关键词：
EAGER Mechanics Guided Multicellular Self

项目摘要

During the development of multicellular tissues, the formation of shape and pattern occur in fairly standard ways. Homogeneous cell populations are triggered to autonomously self-organize into structured, multicellular tissues. However, the signals that guide this organization remain a mystery, and these processes do not consistently occur properly or completely in an a laboratory culture environment. The goal of this EArly-concept Grant for Exploratory Research (EAGER) project is to develop a microfluidic system in which the culture environment can be precisely controlled so that tissue development from stem cells can be directed towards reproducible structures. Correct tissue development requires both biochemical and mechanical signals, which will be precisely controllable through this system. These signals then trigger gene expression within initially homogeneous sets of cells so that they begin a developmental path. The microfluidic system will also be compatible with live imaging, to support future advanced studies into mechanisms driving cellular organization and tissue development. Improved understanding of 3D tissue development into correctly organized structures will significantly enhance research related to developmental biology, including congenital malformations, as well as tissue engineering and regenerative medicine. Educational and outreach plans include integrating high school students, with priority given to women and underrepresented minorities, into the lab for a 4-8 week research experience as well as engaging undergraduates in research. These projects will also provide an opportunity for senior graduate students and post-doctoral fellows to gain mentorship experience. A new course is also being developed related to stem cell biotechnology and regenerative medicine, which will directly draw on the advances in science that support this research. This cross-disciplinary project involves stem cell biology, developmental biology, signal transduction, mechanobiology, and microfluidics. The objectives of this study are to develop a microfluidic device and use it to achieve controllable synthesis of both the epiblast cyst (a columnar tissue) and the amniotic cyst (a squamous tissue) from human pluripotent stem cells. The multi-chambered, microfluidic system will vary both cell loading and chemical induction factors. Cell seeding will be assessed with confocal imaging. Live imaging and immunostaining will be used to monitor mofphogenetic dynamics of the cyst formation, including lumenogenesis (cavity formation) as well as changes in cell shape. Cell fate will be confirmed by staining for epiblast and amniotic lineage markers, and the dynamics of epithelialization will be examined at varying time points.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在多细胞组织的发展过程中，形状和模式的形成以相当标准的方式发生。均质细胞群体被触发以自主自治为结构化的多细胞组织。但是，指导该组织的信号仍然是一个谜，这些过程在实验室文化环境中并不能始终如一地或完全出现。这项对探索性研究的早期概念赠款（急切）项目的目的是开发一个微流体系统，在该系统中可以精确控制培养环境，以便可以将来自干细胞的组织发育指向可再现的结构。正确的组织发育需要生化和机械信号，这将通过该系统准确控制。然后，这些信号在最初均匀的细胞集中触发基因表达，以便它们开始发展路径。微流体系统也将与实时成像兼容，以支持驱动细胞组织和组织发展的机制的未来高级研究。对正确组织结构的3D组织发展的了解将大大增强与发育生物学有关的研究，包括先天性畸形，组织工程和再生医学。教育和外展计划包括将高中生与妇女和代表性不足的少数民族优先融合到实验室中，以获得4-8周的研究经验，并吸引本科生从事研究。这些项目还将为高级研究生和博士后研究员提供一个机会，以获得指导经验。还开发了与干细胞生物技术和再生医学有关的新课程，该课程将直接利用支持这项研究的科学进步。这个跨学科的项目涉及干细胞生物学，发育生物学，信号转导，机械生物学和微流体学。这项研究的目的是开发微流体装置，并使用它来实现来自人类多能干细胞的尤其细胞囊肿（柱状组织）和羊膜囊肿（鳞状组织）的可控合成。多芯片的微流体系统将改变细胞负荷和化学诱导因子。细胞播种将通过共聚焦成像评估。实时成像和免疫染色将用于监测囊肿形成的莫光发育动力学，包括流明发生（腔形成）以及细胞形状的变化。细胞命运将通过染色的表皮细胞和羊膜谱系标记来确认，并且将在不同的时间点进行上皮化的动态。该奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和更广泛的影响审查标准来通过评估来通过评估来支持的。