Mouse pancreas infiltrated immune cell separation method

Abstract

The invention discloses a mouse pancreas infiltrated immune cell separation method which comprises the following steps: digesting pancreas tissue by using mixed digestive juice of collagenase IV, DNAenzyme I and RPMI1640, filtering by using a nylon filtering net of 100 [mu]m, performing density gradient centrifugation, collecting middle-layer cells after centrifugation, and analyzing cell yield,a survival rate and cell types. Results show that (3.0+ / -1.2)*10<5> cells can be collected from each mouse, the survival rate of separated cells is (96.7+ / -2.2)%, flow cytometry shows that the contentof CD45 positive T cells is (97.1+ / -3.5)%, the content of CD3 positive T cells is (61.2+ / -5.7)%, the contents of CD4 positive T cells is (49.1+ / -5.1)%, the content of CD8 positive T cells is (47.2+ / -5.3)%, and the content of CD19 positive B cells is (32.8+ / -4.3)%. The conclusion is that the method is simply, efficient and stable, a great number of cells of a high survival rate and high purity arecollected, and a stable and reliable method is provided for related research on separation of pancreas infiltrated immune cells.

Description

technical field [0001] The present application relates to a method for separating cells in tissues, in particular to a method for separating immune cells infiltrated in mouse pancreas. Background technique [0002] Pancreas is one of the most important organs in the human body, and its physiological functions and pathological changes are closely related to life. In recent years, with the improvement of people's quality of life, people's diet structure has also changed, so more and more pancreatic diseases have appeared. At present, the incidence of pancreatic diseases in my country and even in the world is on the rise. There are more and more reports on pancreatic diseases, such as acute and chronic pancreatitis, idiopathic pancreatitis, autoimmune pancreatitis, and pancreatic cancer, and pancreatic diseases have gradually attracted attention. Many researchers focus on the study of the pancreas on the signal transduction pathways related to pancreatic cancer, the molecular...

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Problems solved by technology

[0003] In view of the shortcomings of the existing separation methods, such as long time-consuming, low yield, complicated steps and inability to be used for infiltrating immune cells in the pancreas, the present invention provides a set of simplified, efficient and stable separation methods for infiltrating immune cells in the mouse pancreas

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