CELL SORTER TO SUPPORT BASIC RESEARCH

支持基础研究的细胞分选仪

• 批准号: RTI-2017-00061
• 负责人: Ma, David
• 金额: $ 10.93万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=616537
• 关键词: CELL; SORTER; SUPPORT; BASIC; RESEARCH

The applicants of this proposal conduct leading edge basic lipid research in fields of physiology, metabolism and cell signalling. Studies to date have addressed questions about how fatty acids, phospholipids, mechanical stress and exercise influence whole body physiology, nutrient metabolism and cell signalling. These fundamental questions have been addressed using a combination of whole body, cell and tissue culture approaches. The vast majority of tissues in the body are comprised of a wide range of cell types. This makes it difficult to determine which particular cells within a given tissue bed are responding to the intervention. Tissue analyses examining gene and protein changes make the basic assumption that all cells isolated are the same, however, this is a gross oversimplification and a multitude of other cell types are also present. For example, these may include stromal and epithelial cells or infiltrating immune cells. Centrifugation and density dependent methods are available but lack specificity and sensitivity, thus requiring large amounts of tissues (i.e. pooling organs from multiple animals) to obtain relatively impure preparations. Similarly, in cell culture approaches it is often desirable to sort cells based on phenotypical characteristics following treatment and then assess the desired experimental endpoints. These experimental needs can only be accomplished using a flow cytometer equipped with a cell sorter, infrastructure that is not readily available to the applicants. An instrument that meets the needs of the investigators and competitively priced is the Bio-Rad S3e cell sorter. This is an entry level workhorse flow cytometer which also has sorting capability. The only other direct competitor is the Sony SH800. However, while comparable in features, it is not competitively priced. Higher end flow cytometers have capabilities far exceeding the routine needs of these investigators and also costing upwards of 1 million. It is anticipated that acquiring this instrument will enhance the direct training of 25+ high quality personnel (per year) across 4 laboratories. Furthermore, it is anticipated that additional faculty and their trainees will seek the use of this instrument once established in the Department of Human Health and Nutritional Sciences at the University of Guelph. In recognition of the importance of this instrument, the Department will provide financial support to support budgetary needs. Overall, the ability to isolate purified populations of cells will enable the elucidation of mechanisms of action and enhance the training environment for many high quality personnel.

该提案的申请人在生理学，代谢和细胞信号传导领域进行前沿基础脂质研究。迄今为止的研究已经解决了脂肪酸，磷脂，机械应力和运动如何影响全身生理，营养代谢和细胞信号传导的问题。这些基本问题已经得到解决，使用整体，细胞和组织培养方法的组合。身体中的绝大多数组织由广泛的细胞类型组成。这使得难以确定给定组织床内的哪些特定细胞响应于干预。检查基因和蛋白质变化的组织分析做出了基本假设，即所有分离的细胞都是相同的，然而，这是一种严重的过度简化，并且还存在许多其他细胞类型。例如，这些可以包括基质和上皮细胞或浸润性免疫细胞。可使用稀释和密度依赖性方法，但缺乏特异性和灵敏度，因此需要大量组织（即汇集来自多个动物的器官）以获得相对不纯的制剂。类似地，在细胞培养方法中，通常期望基于处理后的表型特征对细胞进行分选，然后评估期望的实验终点。这些实验需要只能使用配备有细胞分选器的流式细胞仪来完成，申请人不容易获得这种基础设施。Bio-Rad S3 e细胞分选仪是一种能够满足研究者需求且价格具有竞争力的仪器。这是一个入门级的主力流式细胞仪，也有分选能力。唯一的竞争对手是索尼SH 800。然而，虽然在功能上具有可比性，但它的价格并不具有竞争力。高端流式细胞仪的能力远远超过这些研究人员的常规需求，成本也超过100万美元。预计获得该仪器将加强对4个实验室25名以上高素质人员（每年）的直接培训。此外，预计一旦在圭尔夫大学人类健康和营养科学系建立，将有更多的教员及其受训人员寻求使用这一仪器。认识到这一工具的重要性，该部将提供财政支持以满足预算需求。总的来说，分离纯化的细胞群的能力将能够阐明作用机制，并为许多高素质的人员改善培训环境。