Isolation of Mouse Pancreatic Endothelial Cells.
Publication Year:
2024
PubMed ID:
38975772
Public Summary:
The pancreas plays a crucial role in managing the body’s metabolism by producing important hormones, like insulin, and digestive enzymes. It is filled with tiny blood vessels, called capillaries, which help deliver nutrients and support the pancreas's functions. Special cells, called endothelial cells (ECs), line these capillaries and are essential for the pancreas’s development and health. When these cells don’t work well, it can lead to pancreas-related diseases, such as diabetes and cancer.
Studying pancreatic endothelial cells (pECs) is important for understanding how the pancreas works and how diseases develop. Although mouse models are often used for research, extracting enough pECs from the mouse pancreas has been challenging due to their small numbers and the risk of cell damage from digestive enzymes in the pancreas. To overcome these obstacles, we developed a reliable method to isolate healthy pECs from the mouse pancreas. This new approach combines gentle techniques to separate the cells and a selection process to enrich the ECs, resulting in higher quality cells that can be used for various experiments. This protocol could be useful for research on many diseases and in different mouse models.
Scientific Abstract:
The pancreas is a vital organ for maintaining metabolic balance within the body, in part due to its production of metabolic hormones such as insulin and glucagon, as well as digestive enzymes. The pancreas is also a highly vascularized organ, a feature facilitated by the intricate network of pancreatic capillaries. This extensive capillary network is made up of highly fenestrated endothelial cells (ECs) important for pancreas development and function. Accordingly, the dysfunction of ECs can contribute to that of the pancreas in diseases like diabetes and cancer. Thus, researching the function of pancreatic ECs (pECs) is important not only for understanding pancreas biology but also for developing its pathologies. Mouse models are valuable tools to study metabolic and cardiovascular diseases. However, there has not been an established protocol with sufficient details described for the isolation of mouse pECs due to the relatively small population of ECs and the abundant digestive enzymes potentially released from the acinar tissue that can lead to cell damage and, thus, low yield. To address these challenges, we devised a protocol to enrich and recover mouse pECs, combining gentle physical and chemical dissociation and antibody-mediated selection. The protocol presented here provides a robust method to extract intact and viable ECs from the whole mouse pancreas. This protocol is suitable for multiple downstream assays and may be applied to various mouse models.
