Abstract
Studies on pancreatic cell physiology rely on the investigation of exocrine and endocrine cells in vitro. Particularly, in the case of the exocrine tissue these studies have suffered from a reduced functional viability of acinar cells in culture. As a result not only investigations on dispersed acinar cells and isolated acini were limited in their potential, but also prolonged studies on pancreatic exocrine and endocrine cells in an intact pancreatic tissue environment were unfeasible. To overcome these limitations, we aimed to establish a pancreas tissue slice culture platform to allow long-term studies on exocrine and endocrine cells in the intact pancreatic environment. Mouse pancreas tissue slice morphology was assessed to determine optimal long-term culture settings for intact pancreatic tissue. Utilizing optimized culture conditions, cell specificity and function of exocrine acinar cells and endocrine beta cells were characterized over a culture period of 7 days. We found pancreas tissue slices cultured under optimized conditions to have intact tissue specific morphology for the entire culture period. Amylase positive intact acini were present at all time points of culture and acinar cells displayed a typical strong cell polarity. Amylase release from pancreas tissue slices decreased during culture, but maintained the characteristic bell-shaped dose-response curve to increasing caerulein concentrations and a ca. 4-fold maximal over basal release. Additionally, endocrine beta cell viability and function was well preserved until the end of the observation period. Our results show that the tissue slice culture platform provides unprecedented maintenance of pancreatic tissue specific morphology and function over a culture period for at least 4 days and in part even up to 1 week. This analytical advancement now allows mid -to long-term studies on the cell biology of pancreatic disorder pathogenesis and therapy in an intact surrounding in situ.
Highlights
Malfunction of the exocrine or endocrine pancreas can lead to a number of devastating and life threatening diseases like pancreatitis, pancreatic cancer or diabetes mellitus
Most commonly primary exocrine acinar cells and endocrine beta cells are obtained as dispersed cells by enzymatic digestion of pancreatic tissue followed by exposure to chelating agents [1]
Over a culture period of 7 days tissue slice area significantly decreased to 60.5613.6% on day 4 and 33.8614.6% on day 7, when compared to the initial surface area measured on day 0 (Fig. 1A)
Summary
Malfunction of the exocrine or endocrine pancreas can lead to a number of devastating and life threatening diseases like pancreatitis, pancreatic cancer or diabetes mellitus. Type and characteristics of the cell preparations used in these studies determine which physiological and pathophysiological states can be simulated and which conclusions can be made for in vivo organ function. In the pancreas acinar cells and beta cells are organized in tissue specific structures, the acinus and the islet of Langerhans, respectively. Correct performance of both cell types has been shown to depend on cell to cell contacts established in these functional units [2,3]. Isolated acini and islets of Langerhans have been the preparation of choice when aiming to study pancreatic exocrine and endocrine cell biology in a close to physiological setting in vitro
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