Construction method of pancreatic duct epithelial cell and pancreatic acinar cell co-culture system for simulating in vivo microenvironment

Abstract

The invention discloses a construction method of a pancreatic duct epithelial cell and pancreatic acinar cell co-culture system for simulating in vivo microenvironment, and belongs to the technical field of cell biology. The method comprises the steps of inoculating an upper layer of a Transwell dual-layer culturing chamber with original generation pancreatic acinar cells, inoculating a lower layer of the Transwell dual-layer culturing chamber with human pancreatic duct epithelial cells, performing co-culturing, adding sodium taurocholate in the pancreatic duct epithelial cells at the lower layer of the Transwell dual-layer culturing chamber, and observing the vitality, the shape and the apoptosis of the upper layer pancreatic acinar cells. Damage of the pancreatic duct epithelial cells and the pancreatic acinar cells are combined, a non-contact type co-culture manner is adopted for building the pancreatic duct epithelial cell and pancreatic acinar cell co-culture system. In the co-culture system, the two cells can exchange exocytosis secretion substances through Transwell transmembrane. The system can well simulate the pancreatitis induced by cholestasis, and the invention provides an in vitro cell model which can really reflect pancreatitis induced by cholestasis.

Description

technical field [0001] The invention relates to a method for constructing a co-culture system of pancreatic ductal epithelial cells and pancreatic acinar cells simulating the microenvironment in vivo, and belongs to the technical field of cell biology. Background technique [0002] Acute pancreatitis (AP) is a common digestive system disease characterized by pancreatic parenchymal cell injury and inflammatory cascade reaction. The global annual incidence rate of AP is as high as 13-45 / 100,000 people. Among them, about 80% of acute pancreatitis is a mild self-limiting disease with a good prognosis, but about 20-30% of severe acute pancreatitis continues to progress to systemic inflammatory response syndrome or multiple organ failure, resulting in death. The rate is as high as 10-30%. Biliary origin is the main factor in the occurrence of severe acute pancreatitis, biliary tract stones, inflammation, etc. can cause pancreatic duct obstruction, pancreatic mucosal barrier dama...

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Problems solved by technology

[0004] The exocrine part of the pancreas is composed of acini and ducts. Previous AP-related studies have focused on pancreatic acinar cells. Our study believes that the classic retrograde injection of sodium taurocholate into the pancreaticobiliary duct to establish the AP model, the transient high pressure of the pancreatic duct and exogenous Sexual stimulation will first cause pancreatic ductal epithelial cell damage, pancreatic mucosal barrier damage, pancreatic juice overflow, and then cause abnormal activation of trypsinogen in pancreatic acini to trypsin, leading to pancreatic self-digestion

However, there are no reports on the association of pancreatic acinar cells with ductal cells

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