Amplification method of pancreatic cells and differentiation method and application of pancreatic precursor-like cells

Abstract

The invention provides an amplification method of pancreatic cells. The amplification method comprises the following steps: sequentially carrying out digestion treatment, resuspension treatment, discontinuous density gradient centrifugation treatment and amplification treatment on pancreatic tissues. According to the amplification method disclosed by the invention, pancreatic ducts of mammals are used as sources of pancreatic precursor-like cells, and islet cells and acinar cells in obtained cell clusters are removed after the discontinuous density gradient centrifugal treatment, thus being good for improving the yield of pancreatic precursor-like cells, and avoiding ethical limitations and possible carcinogenic risk caused by the use of embryonic stem cells; and an amplification culture medium used in the invention comprises a reprogramming substance consisting of a plurality of small molecule compounds, so as to avoid the risks of nonspecificity, off-target deletion, etc., which are easily caused by changing gene sequences by using a gene editing method. The invention also provides a differentiation method and application of the pancreatic precursor-like cells obtained by the amplification method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to methods and applications for the expansion and differentiation of pancreatic cells. Background technique [0002] Diabetes mellitus is a chronic metabolic disorder characterized by insufficient insulin secretion or hyperglycemia caused by insulin resistance in surrounding tissues. In the advanced stage, it can lead to damage and dysfunction of tissues and organs throughout the body, especially the eyes, kidneys, cardiovascular system and nervous system. organ failure. Drug therapy and insulin supplementation are currently the most common treatments for diabetes, but they cannot completely cure diabetes, and it is difficult to prevent the development of long-term complications. [0003] Human β-cells can secrete insulin, reduce blood sugar, and regulate fat and protein metabolism. Human embryonic stem cells, because of their unlimited proliferation ability, can theoretically generate...

AI Technical Summary

Problems solved by technology

In the prior art, cells with insulin secretion function can be obtained by gradually inducing embryonic stem cells, but the induction process is complicated, long-term and costly; in addition, in the cell mass induced by embryonic stem cells, cells with insulin secretion function The proportion generally does not exceed 30%, and the later purification process is easy to contaminate the cell mass, so that the obtained cells with insulin secretion function have a potential carcinogenic risk

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