Method for preparing pancreatic single cell suspension of chronic pancreatitis mouse

Abstract

The invention provides a method for preparing a pancreatic single cell suspension of chronic pancreatitis mouse. The method comprises the following steps: A chronic pancreatitis mice model builing; B, chronic pancreatitis mice pancreatic tissue sampling; C, pancreatic tissue collagenase digestion: a mixture of type I and type V collagenase is prepared by using a high glucose DMEM medium, and the collagenase mixture solution is preheated at 37 DEG C before treatment; the pancreatic tissue obtained in step B is transferred to the pre-warmed collagenase mixture, the materials are mixed fully, digested at 37 DEG C for 30 to 90 min, and mixed thoroughly every 10 min; and D, pancreatic single cell separation: the material is filtered once with a 70 [mu]m filter, the residue is discard, the cell suspension is centrifuged, a supernatant is discard, and resuspend deposition is carried out in high glucose DMEM medium; a 30 [mu]m filter is used for refiltering again, the residue is discarded, the cell suspension is centrifuged, and the supernatant and the upper cell debris, a DNA flocculent twister and a middle layer of blood cells are discarded, the white parenchymal cells at a lowermost layer are retained, and a PBS solution is resuspended to obtain the pancreatic single cell suspension of chronic pancreatitis mouse.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a preparation method, construction method and application of a mouse pancreas single cell suspension with chronic pancreatitis. Background technique [0002] The pancreas is a hidden organ in the abdomen. It performs important functions such as food digestion and blood sugar regulation. Its cellular components are relatively complex, including pancreatic acinar cells, stellate cells, endocrine cells, endothelial cells, ductal cells, and lymphocytes. Pancreatic cell analysis is an effective method to understand the mechanism of action of the pancreas. Traditional pancreatic cell sequencing and analysis are all samples taken from the whole or part of the pancreas tissue. With the development of sequencing technology, single-cell sequencing, as a high-throughput sequencing method, can not only reveal the precise mechanism of individual cells, but also analyze the differences bet...

AI Technical Summary

Problems solved by technology

[0006] However, the pancreas in chronic pancreatitis undergoes atrophy and fibrosis, and parenchymal cells are surrounded by collagen fibers, which are very different from normal pancreatic cell morphology

Digestion of normal pancreatic cells is difficult to achieve separation of individual cells and high cell viability, so it is impossible to obtain a single cell suspension of pancreas in chronic pancreatitis

Through searching, there is currently no method for preparing single cell suspension for CP mouse model

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