Cell culture room for 2D-3D in vitro systems

我们请求资金更换和升级我们细胞培养设施的设备。本次申请购买的项目有四项：2台培养箱，1台立体显微镜用于解剖，1台倒置显微镜用于观察和评价培养物。该设备将支持申请人、共同申请人和合作者的研究项目。当前应用中涉及的nserc - dg资助的研究项目需要使用可靠且功能强大的培养设施。不幸的是，目前情况并非如此。我们从以前的研究人员那里继承了我们的细胞培养设备，尽管这些设备有效地服务了30多年，但它们出现了故障，已经过时了。细胞培养是基础科学和翻译研究问题的一个通用和强大的工具。细胞培养帮助我们了解细胞生物学、组织形态、疾病机制、药物作用、蛋白质生产和组织工程的发展。目前研究中使用的大多数培养方法都是基于二维（2D）细胞培养单层。二维细胞培养的主要优点是易于获得和简单的环境控制，不受限制的细胞观察，直接进入细胞，包括易于测量和操作。然而，传统的2D细胞培养很难模拟生物体中的条件，其中细胞在复杂的三维（3D）微环境中生长，需要持续的血管灌注来提供氧气和营养物质，并去除代谢废物。此外，其他因素，如细胞形态的改变、极性的丧失和细胞分裂参数的改变，限制了基于二维培养系统的研究范围。这些限制导致了替代性体外模型系统的发展，旨在重现更接近细胞在体内经历的条件。其中一种方法是3D培养，其表现出更接近活体条件的特征。当前RTI应用支持的培养工作将包括二维单层培养，例如从小鼠海马、感觉神经节和细胞系中产生神经元原代培养；以及3D培养系统，包括气液界面（ALI）的气道上皮细胞和胃肠道类器官，包括与感觉神经元共培养的类器官。通过这项拨款请求的设备将支持四位NSERC-DG资助的研究人员的研究项目。这笔拨款中包含的设备代表了细胞培养设施的“工作马”，我们的研究项目，无论是单独的还是合作的，都严重依赖于这些设备的正常工作。RTI的支持将加强nserc资助的四个实验室的研究活动，包括在所需设备的工作寿命期间培训数百名HQP。

We are requesting funds to replace and upgrade equipment in our cell culture facility. The present grant application aims to purchase four items: two culture incubators, a stereo microscope for dissections, and an inverted microscope for observation and evaluation of cultures. This equipment will support the research programs of the applicant, co-applicant, and collaborators. The NSERC-DG-funded research programs involved in the current application require access to a reliable and highly functional culture facility. Unfortunately, that is not currently the case. We inherited our cell culture equipment from previous researchers, and even though these pieces of equipment served fruitfully for 30+ years, they are malfunctioning and are obsolete. Cell culture is a versatile and robust tool in basic scientific and translation research questions. Cell cultures help us to understand cell biology, tissue morphology, and mechanisms of diseases, drug action, protein production and the development of tissue engineering. Most of the culture methods utilized in research today are based on two-dimensional (2D) cell culture monolayers. The main advantages of 2D cell cultures are the accessibility and simplicity of environmental control, unrestricted cell observation, direct cell access including the easiness for measurement and manipulation. However, conventional 2D cell cultures poorly mimic the conditions in the living organism, in which cells grow within a complex three-dimensional (3D) microenvironment, with continuous vascular perfusion required to provide oxygen and nutrients, and to remove metabolic waste products. Furthermore, other factors such as changes in cell morphology, loss of polarity, and changed cell division parameters, limit the scope of research based on 2D culture systems. These limitations led to the development of alternative in vitro model systems aimed at recreating conditions closer to those that cells experience in vivo. One such method is 3D cultures, which exhibit features closer to the in vivo conditions. The culture work that will be supported by the current RTI applications will include 2D culture monolayers, such as the generation of neuronal primary cultures form hippocampus, sensory ganglia from mice, and cell lines; as well as 3D culture systems including airway epithelial cells at the air-liquid interface (ALI) and gastrointestinal organoids, including organoids co-cultured with sensory neurons. The equipment requested through this grant will support the research programs of four NSERC-DG funded researchers. The pieces of equipment included in this grant represent the "work horses" of a cell culture facility, our research programs, both individually and collaboratively, critically depend on having these pieces of equipment in working order. The RTI support would enhance research activities at four NSERC-funded laboratories, including the training of hundreds of HQP through the work life of the equipment requested.