Advanced cell sorting technology to properly investigate biological mechanisms in the presence of cellular heterogeneity

先进的细胞分选技术可在存在细胞异质性的情况下正确研究生物机制

• 批准号: RTI-2023-00145
• 负责人: Verschoor, Chris
• 金额: $ 10.93万
• 依托单位: Laurentian University of Sudbury
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748504
• 关键词: Advanced; cell; sorting; technology; properly

In studies of the biological mechanisms governing cellular responses to their microenvironment it is common to take one or more individual measurements to characterize a given response. These measures effectively present the average of every cell within that population being studied. For this reason, it is critical that the cellular population being investigated is homogenous; in other words, every cell should be very similar to one another and their responses to a stimulus should be similar as well. If not, the resultant measurement may not accurately represent the phenomenon being studied unless those heterogenous cells are separated from one another. In the case where a cell's microenvironment or some other form of exogenous stimuli drives that cell to differentiate into one or more new, highly specialized cell types, heterogeneity is expected. Here, the need to separate or sort cells according to their differentiated phenotype is of utmost importance, as it will allow investigators to use their powerful toolkit of cellular and molecular techniques and measures to understand the biological underpinnings of those differentiated subsets. Clearly, to test novel and innovative hypotheses in cellular and molecular biology, advanced instrumentation to physically sort and separate heterogenous cell types into defined populations is fundamental. Unfortunately, for our research team representing NSERC-funded investigators from Laurentian University, the Northern Ontario School of Medicine University and Health Sciences North Research Institute, there are no adequate means to achieve this goal within a 400 km radius. With this funding, we aim to purchase an automated cell sorting instrument that will allow us to isolate up to two phenotypically distinct types of cells from a mixed population, which we can then analyze using traditional cellular and molecular biology techniques. This will greatly advance our research goals, allowing our investigators to study signalling mechanisms of unique cell subsets that they are currently unable to obtain; use cutting edge CRISPR-based screening tools to obtain rare genetic mutant cells that may uncover the nature of neoplastic cell transformation; and sort out cells within a seemingly homogenous population that for some yet to be discovered reason are able to resist the cancer causing effects of radiation. Furthermore, adding such advanced instrumentation to our current comprehensive system of cell biology tools will enrich the highly-qualified training environment that we provide to students and other trainees in Northeastern Ontario. This will improve the marketability of their skills, while providing regional natural sciences, biotechnology and medical industries better candidates to drive innovation in their businesses.

在控制细胞对其微环境的反应的生物学机制的研究中，通常采取一个或多个单独的测量来表征给定的反应。这些测量有效地呈现了正在研究的群体中每个细胞的平均值。因此，研究的细胞群体必须是同质的;换句话说，每个细胞都应该彼此非常相似，它们对刺激的反应也应该相似。如果不是，则所得的测量结果可能不能准确地代表所研究的现象，除非这些异质细胞彼此分离。在细胞的微环境或一些其他形式的外源刺激驱动该细胞分化成一种或多种新的、高度特化的细胞类型的情况下，异质性是预期的。在这里，根据细胞的分化表型对细胞进行分离或分类的需要是至关重要的，因为它将使研究人员能够使用其强大的细胞和分子技术工具包和措施来了解这些分化亚群的生物学基础。 显然，为了测试细胞和分子生物学中的新的和创新的假设，将异质细胞类型物理分选和分离到定义的群体中的先进仪器是至关重要的。不幸的是，对于我们的研究团队代表NSERC资助的研究人员从劳伦特大学，北方安大略医学院和健康科学北方研究所，有没有足够的手段来实现这一目标在400公里半径。有了这笔资金，我们的目标是购买一种自动化细胞分选仪器，使我们能够从混合群体中分离出两种表型不同的细胞类型，然后我们可以使用传统的细胞和分子生物学技术进行分析。这将极大地推进我们的研究目标，使我们的研究人员能够研究他们目前无法获得的独特细胞亚群的信号传导机制;使用基于CRISPR的尖端筛选工具获得罕见的遗传突变细胞，这些细胞可能揭示肿瘤细胞转化的性质;并在一个看似同质的群体中挑选出细胞，这些细胞由于某些尚未发现的原因能够抵抗辐射的致癌作用。此外，将这种先进的仪器添加到我们目前全面的细胞生物学工具系统中，将丰富我们为安大略东北部的学生和其他学员提供的高质量培训环境。这将提高他们技能的市场化程度，同时为区域自然科学、生物技术和医疗行业提供更好的候选人，以推动其业务创新。