The following are the awardees of the IIDP Technology Resource Award (TRA), funded to advance human islet research by providing support for high-impact studies that will enhance services provided by IIDP to the research community.

Name and Institution	Project Title and Years	Project Abstract
Juan Dominguez-Bendala, PhD University of Miami Miller School of Medicine	Optimization of Human Pancreatic Slice Procurement, Culture and Distribution 0000-0001-9410-9143 2024 - 2025	Human pancreatic slices (HPSs) have rapidly become a model of reference for the study of the endocrine-exocrine cross talk in the normal biology and pathological conditions of the pancreas. These live sections of the organ exhibit minimal disruption of the extracellular matrix, maintaining not only the native proportions of the different cell types that make up the pancreas, but also the cell-to-cell interactions and the physical microenvironment of islets, acini, and ducts. Our team recently developed conditions to extend the viability and function of slices beyond 10 days of culture. This technical accomplishment has opened the door to the long-term study of pancreatic disease in an in vitro setting closely resembling the real human organ. Pioneered by the Network of Pancreatic Organ Donors with diabetes (nPOD), the distribution of viable/functional human slices shipped from centralized procurement centers to individual investigators around the globe has become a much sought-after initiative in the field. However, there is dearth of centers with the ability to generate slices in-house, and sample availability is limited. Additionally, there is still a widespread perception that research on HPSs is too specialized or beyond the reach of most labs. Our proposal is geared toward the refinement and standardization of distribution, culture, and analytical approaches that may facilitate the goal of expanding the availability of this model to any interested team. We present here an integrated plan to make this biological material available to the diabetes community at large, along with optimized and validated protocols to culture long-term, measure viability and function (exocrine and endocrine), and conduct viral transduction/dissociation for a range of applications. In particular, we propose the following specific aims: (1) Optimization of viability and function of HPS samples processed from tissues originated from IIDP islet isolation centers. (2) Optimization of shipment of viable slices to destination labs; and (3) Optimization of long-term culture, viral transduction and dissociation of HPSs for FACS/Sorting/scRNAseq applications. We present here a detailed plan to manage the conduct of these three specific aims from the same 12 HPS preparations (one/month), thus maximizing the utilization of this precious biological material within the constraints of a one-year project. The pioneering expertise of our team in this field, along with our strategic partnership with established IIDP centers across the US are major strengths of our proposal.
Rebecca Hull-Meichle, PhD Alberta Diabetes Institute	Strategies to Optimize Isolation of Ductal Epithelial Cells from Human Pancreas 0000-0001-9690-4087 2024 - 2025	This IIDP Technology Resource Award is focused on strategies to isolate, characterize and propagate ductal epithelial cells from IIDP human pancreas preparations. This work will benefit research focused on exocrine pancreas function, mechanisms of pancreas disease (e.g. pancreatitis, cystic fibrosis or pancreatic cancer) and interactions between the exocrine and endocrine pancreas. The isolation of pancreatic ductal epithelial cells (PDECs) has been reported by several groups, including our own (see preliminary data) but a simple, consistent, scalable method for use across IIDP centers has not been established and is the goal of this proposal. In Aim 1, we will compare isolation of PDECs from two tissue sources: First, we will use acinar preparations, prepared per IIDP protocol, which contain abundant PDECs, and which we and others have used to date. Second, we will establish culture conditions to encourage propagation of PDECs from the ductal tree that remains at the end of the islet isolation procedure. We have already demonstrated efficacy of this approach on a small scale (see preliminary data) and will expand this in the studies in Aim 1. We will test the ability of these cells to be propagated in 2D and 3D culture and test their ability to be frozen, shipped and recovered from cryostorage. In Aim 2, we will thoroughly characterize the cells isolated using both approaches by flow cytometry, qPCR and immunohistochemistry. In doing so, we will compare PDEC markers, along with the residual presence of other cell types (principally acinar cells) in cells isolated with each approach, both within- and between donors. We will also monitor expression of PDEC markers, as well as re-expression of progenitor markers in cells cultured over multiple passages. We will also screen dyes, starting with the agglutinin DBA, for efficacy in identifying human PDECs, in an analogous way to how dithizone is used to aid in islet identification. The goal of this aim is to provide IIDP and the research community with a simple, reproducible tool to identify PDECs and/or assess their viability/function. In addition to providing IIDP users with improved access to PDECs, this work will also accelerate our own research into PDEC pathology in cystic fibrosis and how this contributes to the pathogenesis of cystic fibrosis-related diabetes.
Ian Sweet, PhD University of Washington School of Medicine, Diabetes Institute	Concomitant Metabolic and Functional Analysis of Multiple Tissue Types 0000-0002-7565-1663 2024 - 2025	There is a critical need for the development and validation of standardized protocols and assays to measure quality, viability and function of islets and associated tissue that are provided by the IIDP. We have developed a fluidics system that meets these requirements and importantly is easy to use and has good concordance between laboratories, requisite attributes for a gold standard. Importantly, the system is able to control the concentration of dissolved O2 in contact with the islets in the perifusion chamber. The focus of the grant is on islet function and the use of the system to evaluate and study isolated human islets. However, consistent with the RFA, we also will demonstrate the system's ability to maintain and assess other tissues, as well as the identifying the optimal dissolved O2 concentration in the perifusate. To do this, pancreatic tissue slices will be used both as an important tissue model, where islets may reside in a state that may be closer to their native state than isolated islets, but also as representing a larger piece of tissue that may present an increased challenge to provide adequate oxygenation. Historically, perifusion systems have been technically challenging and for the most part static culture analysis has been used to evaluate islets and other tissue. However, the system we have developed has no moving parts or pumps, is easier to use than the static analysis, and also has many advantages. Most notably, islets and other isolated tissue samples are highly energetic and demand a steady supply of oxygen, conditions that are better met with flow systems. The first aim of the grant, done in collaboration with Drs. Juan Dominguez-Bendala and Julia Panzer, whose laboratories have experience in processing and studying pancreatic tissue slices, takes advantage of our system's ability to select levels of dissolved O2, which will allow us the determine the adequacy of the delivery rate of O2 to islets and pancreatic tissue slices during assessment. In the second aim, the system will be used to compare islets from control subjects vs. those with T2D. In the third aim, two additional fluidics systems have been sent to collaborating institutions (Drs. Carmella Evans Molina (Indiana University) and Marcela Brissova (Vanderbilt) and the results of performing the same protocols on the same batches of islets will be assessed for inter-laboratory variability. In summary, this pilot and feasibility grant will, with a relatively small number of experiments, demonstrate the ability of the flow system to meet multiple critical needs of the IIDP network including measurement of bioenergetics and secretory function of islets and slices and their dependence on O2 concentration.