Non-contact plotter for precise patterning of proteins on novel microdevices

体内的细胞对机械力、化学信号和它们所依附的蛋白质基质作出反应。干细胞工程策略试图利用这些信号引导非特化祖细胞成为可用于治疗的功能性细胞。为了使这种方法起作用，我们需要知道适当的信号组合，以使细胞产生我们想要的功能细胞类型。我们的跨学科团队正在开发新技术来筛选机械、化学和蛋白质信号的组合，以寻找它们在引导干细胞分化方面的综合作用。这些微型设备使我们能够以比目前更高的速率和更低的成本同时测试许多样品上的多种调节因子组合。我们目前的设备对细胞施加精确的机械和化学刺激，但我们很难创造出精确的基质蛋白质模式来粘附细胞。这是因为标准的蛋白质印刷方法再现性差，图案均匀性差。在这个提案中，我们要求一个自动液体分配器在我们的微型设备上精确的位置打印蛋白质。这种蛋白质打印机可以高速打印，具有极高的精度和分辨率，并且不需要接触微型设备，这对于防止损坏精致的聚合物表面非常重要。打印是由机器人控制的，可以打印任何想要的图案。该打印机的准确性，可重复性和多功能性将使我们能够在我们的微型设备上可靠地以定义良好的模式打印蛋白质。这种能力对我们系统的未来发展至关重要。我们预计，随着进一步的发展，我们的微系统将加速再生医学和干细胞工程领域的发现和创新。该打印机将被安置在多伦多大学一个新的、最先进的生物微器件制造和表征设施中。由于该仪器易于扩展和适应许多应用，并且可以通过我们国家独特的制造设备完全访问，我们预计它将被我们机构和其他机构的几个研究小组广泛使用。

Cells in the body respond to mechanical forces, chemical signals, and the protein matrix to which they are adhered. Stem cell engineering strategies attempt to use these signals to guide unspecialized progenitor cells to become functional cells that can be used therapeutically. In order for this approach to work, we need to know the appropriate combination of signals to give the cells to produce the functional cell types we want.     Our interdisciplinary team is developing new technologies to screen combinations of mechanical, chemical, and protein signals for their integrated effects on guiding stem cell differentiation. These microscale devices allow us to test multiple combinations of regulatory factors on many samples simultaneously at a higher rate and lower cost than is possible currently.  Our current devices apply precise mechanical and chemical stimuli to cells, but we have had difficulty creating precise matrix protein patterns to adhere the cells. This is because standard protein printing methods have poor reproducibility and poor pattern uniformity.     In this proposal, we have requested an automated liquid dispenser to print proteins at precise locations on our microdevices. This protein printer prints at high speed, with excellent precision and resolution, and without contacting the microdevices, which is important for preventing damage to the delicate polymer surfaces. Printing is controlled by a robot that can print any pattern desired. The accuracy, reproducibility, and versatility of this printer will enable us to reliably print proteins in well-defined patterns on our microdevices.  This ability is critical to future development of our systems. We anticipate that with further development, our microsystems will speed discovery and innovation in the fields of regenerative medicine and stem cell engineering.     The printer will be housed at the University of Toronto in a new, state-of-the-art facility for biomicrodevice fabrication and characterization. Because the instrument is readily expandable and adaptable for many applications, and will be fully accessible through our nationally-unique fabrication facility, we expect that it will be used extensively by several research groups from our institution and others.