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    Biomanufacturing

    (C-84) Pancreatic Slices on a Chip as a tool for Type 1 Diabetes Research

    Friday, October 13, 2023
    3:30 PM – 4:30 PM PDT
    Location: Exhibit Hall - Row C - Poster # 84
    Introduction:: In type 1 diabetes, autoimmune destruction of pancreatic β-cells leads to insulin deficiency and loss of glycemic control. Functional long-term cultures of human pancreatic slices allow for the longitudinal tracking of β-cell function, such as insulin secretion, and regeneration from progenitor cells in healthy and diseased tissues. However, current methods for long-term cultures of pancreatic slices utilize static cultures on oxygen permeable dishes which require frequent media changes and result in the buildup of harmful digestive enzymes. To address the limits of a static pancreatic slice culture, we have developed a micro-physiological system (MPS) that can sustain human pancreatic slice cultures for extended periods.  

    Materials and Methods:: We are adapting a previously developed pancreatic islet on a chip platform to create a fluidic chip that allows for continuous media perfusion, preventing the build-up of cellular metabolites and allowing for the sequential administration of biological interrogators providing a more physiologically relevant environment than a static culture. Our Pancreatic Slice on a Chip is capable of sustained slice culture with concurrent fluorescence microscopy, allowing for serial visualization of cellular changes in response to biologic interrogators. To minimize movement artifacts under long-term imaging, a 3D printed bioinert anchor was developed to stabilize slices. The ability to sequentially stimulate the slices with biochemical agents—such as glucose and carbachol— within a microscope allows for detailed glucose-stimulated insulin secretion (GSIS) Ca2+ imaging. As an undergraduate researcher, I will describe my contributions in controlling media flow in our chip (generated by a constant-pressure pump that pressurizes media containers connected to the chip), addition of biological stimulants, and deployment of an incubation chamber that was adapted to have fluidic ports to feed into the chip without breaking the sterility or incubation function. 

    Results, Conclusions, and Discussions:: Within this novel system, we are able to maintain slice cultures under dynamic oxygen levels for 5 days, perform sequential GSIS studies on singular pancreatic slices, and take high-resolution time-lapse images. Under variable media oxygenation levels, we have observed apparent differences in slice GSIS response. Our pancreatic slice on a chip allows for continuous visualization of β-cells, their neogenesis from progenitor cells within a physiomimetic environment and cellular signaling in response to stimuli. 

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