CAREER: Chemomechanical imaging and engineering of single cell phenotype

本研究的总体目标是解决一个尚未解决的生物工程挑战：我们能否通过识别、成像和控制动态细胞表面与相邻材料环境之间的相互作用，来设计细胞表型，使其对人类有利？这个问题是体外和体内功能组织合成的核心。干细胞和祖细胞，部分地由在环境线索下采用几种不同表型的能力所定义，是这种工程功能的特别有利的靶标。目前的局限性部分是由于我们对细胞动态响应环境的一般机制缺乏了解。通过细胞表面特征评估细胞表型的能力对于引导细胞功能的环境的工程化至关重要，特别是在机械响应系统如脉管系统中。因此，本研究的目的是开发一种化学力学方法，使用扫描探针显微镜的先进技术（称为功能化力成像（FFI））来绘制个体（1）血管内皮细胞和（2）成人骨髓间充质干细胞表面的实时变化。连续的图像采集将使配体结合动力学和细胞反应的实时体外分析成为可能。实时识别和量化单个活细胞内细胞表面环境相互作用机制的分子基础的能力将提供一种新的手段，通过这种手段可以识别、分离和分析低频存在的细胞对环境刺激的反应。该提案的教育和社会目标被整合到拟议的研究方法和应用中。PI将通过开发新的本科实验室模块和积极参与外展计划来培训未来的研究人员。

Krystyn J. VanVliet0644846The overall objective of this research is to address an unresolved bioengineering challenge: Can we engineer cell phenotype to human advantage through identification, imaging, and control of interactions between the dynamic cell surface and adjacent material environment? This question is central to synthesis of functional tissue in vitro and in vivo. Stem and progenitor cells, defined in part by the ability to adopt several distinct phenotypes under environmental cues, are particularly advantageous targets of this engineered function. Current limitations are due in part to our poor understanding of the general mechanisms by which cells respond dynamically to the environment. The capacity to assess cell phenotype through cell surface characteristics is critical to the engineering of environments that will guide cell function, particularly in mechanically responsive systems such as the vasculature. Thus, the objective of this research is to develop a chemomechanical approach to map the real-time changes in the surface of individual (1) vascular endothelial and (2) adult bone marrow-derived mesenchymal stem cells using an advanced technique of scanning probe microscopy, termed functionalized force imaging (FFI). Sequential image acquisition will enable real-time, in vitro analysis of ligand-binding kinetics and cell response. The capacity to identify and quantify in real-time the molecular basis of cell surface environment interaction mechanisms within individual living cells will provide a new means by which cells present in low frequency can be identified, isolated, and analyzed in response to environmental stimuli. The educational and societal goals of this proposal are integrated within the proposed research methods and applications. The PI will train future researchers by developing new undergraduate laboratory modules and actively participating outreach programs.