EAGER: Mechanics-Guided Multicellular Self-Organization

在多细胞组织的发育过程中，形状和图案的形成是以相当标准的方式进行的。同质细胞群被触发自主地自组织成结构化的多细胞组织。然而，指导这个组织的信号仍然是一个谜，这些过程在实验室培养环境中并不总是正确或完全发生的。这个早期概念探索性研究奖助金(AGER)项目的目标是开发一种微流体系统，在该系统中可以精确地控制培养环境，从而使干细胞的组织发育可以定向到可复制的结构。正确的组织发育需要生化和机械信号，通过这个系统可以精确地控制这些信号。然后，这些信号在最初同质的细胞组中触发基因表达，从而开始发育过程。微流控系统还将与实时成像兼容，以支持未来对驱动细胞组织和组织发育的机制的高级研究。更好地理解3D组织发育成正确组织的结构将大大促进与发育生物学相关的研究，包括先天性畸形，以及组织工程和再生医学。教育和外展计划包括让高中生进入实验室进行4-8周的研究体验，以及让本科生参与研究，优先考虑女性和代表性不足的少数族裔。这些项目还将为高级研究生和博士后研究员提供获得指导经验的机会。一门与干细胞生物技术和再生医学相关的新课程也正在开发中，它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一种微流控装置，并使用它来实现从人类多能干细胞中可控地合成上胚囊(柱状组织)和羊膜囊(鳞状组织)。多室微流控系统将改变细胞负荷和化学诱导因素。细胞播种将通过共聚焦成像进行评估。将使用实时成像和免疫染色来监测包囊形成的动态变化，包括管腔形成(空洞形成)以及细胞形状的变化。细胞命运将通过上胚层和羊膜谱系标记的染色来确认，上皮化的动态将在不同的时间点进行检查。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.