Growing pancreatic organoids for hereditary pancreatitis

Organoids hold many advantages over two-dimensional cell lines and animal models, allowing for the study of disease that would previously be challenging to accurately reproduce. As such, organoids have accelerated drug development and screening for diseases such as cystic fibrosis and various cancers. We have grown organoids for research into hereditary pancreatitis, a chronic inflammatory condition caused by a series of pancreatitis associated genes. Animal models can struggle to recapitulate this disease due to discrepancies in these genes, such as PRSS1 and SPINK1, indicating the need for a more physiologically relevant model. Pancreatic organoids were grown in a Matrigel matrix from normal tissue samples and characterised based on morphology. A complex growth media is utilised when cultivating these organoids, including Advanced DMEM/F12, Glutamax, HEPES, B-27, N-acetylcysteine, Wnt3a, Rspo1, nicotinamide, gastrin 1, EGF, FGF-10, Noggin, Prostaglandin E2, Y27632 (ROCK inhibitor), and CHIR99021 (GSK-3 inhibitor). The organoids were treated with other gene inhibitors and growth factors such as A83-01, DAPT and NRG1 in order to abate the process of acinar and ductal metaplasia and thus provide a more accurate model of the cell subset most relevant to hereditary pancreatitis. Over 100 tissue samples have been taken across a number of donor pancreases to grow these organoids from healthy tissue. We currently have 152 hereditary pancreatitis patients with known gene mutations in South Australia. Tissue samples will be sourced from these patients undergoing total pancreatectomy with islet auto-transplantation, allowing for the growth of organoids which can recapitulate the disease phenotype based on a patient’s unique combination of mutations. Pancreatic organoids grown from the tissue samples of normal individuals will form a baseline for studies in individuals with HP. Developing organoid models will be key for the development of effective pharmacological and genetic therapies for pancreatitis and pancreatic cancers.