3-D Optical Tracking of Bone Marrow Derived Skin Stem Cells

骨髓源性皮肤干细胞的 3D 光学追踪

• 批准号: 0852658
• 负责人: Stephen Boppart
• 金额: $ 30.6万
• 依托单位: University of Illinois at Urbana-Champaign
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-15 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0852658&HistoricalAwards=false
• 关键词: Optical; Tracking; Bone; Marrow; Derived

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)0852658 Boppart Skin is our largest organ, serving critical roles in fluid homeostasis, thermoregulation, immune surveillance, and self-healing. Disease and/or the loss of major portions of a human?s skin can be disabling and potentially life threatening, and is a major health problem in the U.S. and throughout the world. Stem cells play a critical role in repairing and regenerating many tissues, including the skin. Elucidating the roles that stem cells play in the skin will therefore have a significant impact on not only our fundamental understanding of stem cell dynamics, but also on our treatment of skin diseases, for replacing skin in medical applications, or in rejuvenating skin in our aging population. Recent advances in optical imaging techniques offer an unprecedented combination of high spatiotemporal imaging resolution that can now be applied to visualizing the complex three-dimensional (3-D) dynamics of skin stem cells within normal skin, and in response to skin injury and skin replacement, such as after grafting engineered skin replacements. The intellectual merit of this proposal is represented for the first time by an advanced optical biomedical imaging approach for elucidating the complex dynamics of skin stem cells in vivo and within engineered skin grafts. The hypothesis of this research is that optical coherence and multi-photon microscopy, in an integrated platform, can uniquely track and quantify the different dynamic 3-D in vivo stem cell behaviors in and around autologous and allogeneic engineered skin grafts. With the recent discovery of the skin stem cell niche located within the bulge region of hair follicles, many questions arise as to the dynamic behavior of these stem cells as they migrate from the bone marrow and into the skin niche, as well as in and out of the niche in response to skin injury and disease. This project therefore has intellectual merit in four areas. First, an advanced integrated microscope capable of simultaneous optical coherence and multi-photon microscopy will be utilized to uniquely visualize the structural and functional relationships of stem cells within in vivo skin. Second, this project investigates and longitudinally images in 3-D the migration patterns and dynamics of skin stem cells. This will provide fundamental insight into the role they play in maintaining the function and health of skin. Third, the effects of skin injury, induced by the placement of an autologous skin graft (skin punch biopsy), will be investigated, providing insight into the stem cell dynamics in the healing response. Fourth, this project will longitudinally image the stem cell and tissue responses in vivo following the grafting of allogeneic engineered skin constructs, contributing significantly to the understanding of how skin stem cells interact with engineered tissue grafts within biological hosts. The development and application of more quantitative imaging techniques to analyze the dynamics at the single-cell or cell-population levels will provide further insight into the ability to understand the role of skin stem cells, and ultimately provide a better approach for the treatment of human pathologies that require skin grafting. Taken together, this project is novel in each of these four areas, and the use of these advanced imaging techniques to carry out these investigations is transformative for the fields of stem cell biology and tissue engineering.Considering the broader scope, the outcome of this project is likely to have a significant and broad impact on both the fundamental and clinical understanding on how stem cells behave dynamically in vivo. This project addresses major challenges in stem cell biology and tissue engineering: how to visualize and track cells and small populations of cells in vivo, in 3-D, and longitudinally over time in highly-scattering engineered and natural tissues.This project will integrate state-of-the-art research with educational elements to advance discovery and promote teaching, training, and learning. Undergraduate students, in addition to graduate students, will complete theses related to this work. These students, as well as post-doctoral fellows and research scientists, will develop lifelong career skills in optics, image processing, cell and tissue culture and biology, and the use of pre-clinical models. Under-represented groups including women and minorities will be targeted for research opportunities, and annual laboratory and campus-wide open-house events will be held for outreach to K-12 and community groups. The results and image databases from this project will be disseminated widely through our educational website, local and national conferences, and leading scientific, engineering, and medical publications.

该奖项是根据2009年美国复苏和再投资法案（公法111-5）0852658资助的。皮肤是我们最大的器官，在体液平衡、体温调节、免疫监视和自我修复方面发挥着关键作用。疾病和/或人体主要部分的丧失？美国人的皮肤可能会致残，甚至可能危及生命，是美国乃至全世界的一个主要健康问题。干细胞在包括皮肤在内的许多组织的修复和再生中起着关键作用。因此，阐明干细胞在皮肤中的作用将不仅对我们对干细胞动力学的基本理解产生重大影响，而且对我们治疗皮肤病、在医学应用中替代皮肤或在老龄化人口中使皮肤恢复活力也有重大影响。光学成像技术的最新进展提供了前所未有的高时空成像分辨率组合，现在可以应用于可视化正常皮肤内皮肤干细胞的复杂三维（3-D）动态，以及对皮肤损伤和皮肤替换的反应，例如移植工程皮肤替换后。这一建议的知识价值首次通过先进的光学生物医学成像方法来阐明体内和工程皮肤移植物中皮肤干细胞的复杂动力学。本研究的假设是，光学相干和多光子显微镜，在一个集成的平台上，可以唯一地跟踪和量化不同的动态三维体内干细胞在自体和异体工程皮肤移植物及其周围的行为。随着最近在毛囊凸起区域发现皮肤干细胞生态位，这些干细胞从骨髓迁移到皮肤生态位以及在皮肤损伤和疾病中进出生态位的动态行为引发了许多问题。因此，该项目在四个方面具有智力价值。首先，一个先进的集成显微镜能够同时光学相干和多光子显微镜将被用来独特地可视化体内皮肤内干细胞的结构和功能关系。其次，本项目研究并纵向三维成像皮肤干细胞的迁移模式和动态。这将为他们在维持皮肤功能和健康中所起的作用提供基本的见解。第三，将研究自体皮肤移植（皮肤穿刺活检）引起的皮肤损伤的影响，从而深入了解愈合反应中的干细胞动力学。第四，本项目将对同种异体工程皮肤移植后的干细胞和组织在体内的反应进行纵向成像，这将有助于理解皮肤干细胞如何与生物宿主内的工程组织移植物相互作用。更多定量成像技术的发展和应用，以分析单细胞或细胞群体水平的动态，将进一步深入了解皮肤干细胞的作用，并最终为需要皮肤移植的人类病理治疗提供更好的方法。总的来说，这个项目在这四个领域都是新颖的，使用这些先进的成像技术来进行这些研究对于干细胞生物学和组织工程领域来说是革命性的。考虑到更广泛的范围，这个项目的结果可能会对干细胞在体内动态行为的基础和临床理解产生重大而广泛的影响。该项目解决了干细胞生物学和组织工程中的主要挑战：如何可视化和跟踪细胞和小群细胞在体内，在3-D，纵向随时间在高散射工程和自然组织。该项目将把最先进的研究与教育元素结合起来，以推进发现，促进教学、培训和学习。除研究生外，本科生将完成与本工作相关的论文。这些学生，以及博士后研究员和研究科学家，将在光学，图像处理，细胞和组织培养和生物学以及临床前模型的使用方面发展终身职业技能。包括妇女和少数民族在内的代表性不足的群体将成为研究机会的目标，并将举行年度实验室和校园开放日活动，向K-12和社区团体推广。该项目的结果和图像数据库将通过我们的教育网站、地方和国家会议以及主要的科学、工程和医学出版物广泛传播。