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

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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.