A method for obtaining pancreatic precursor cells and islet β cells by differentiating human pluripotent stem cells

Abstract

The invention discloses a method for obtaining pancreatic precursor cells and islet beta cells by differentiation of human pluripotent stem cells, and relates to the technical field of application of stem cell differentiation in medical treatment, comprising the following steps: preparing human pluripotent stem cells into multifunctional cell spheres culturing the obtained pluripotent cell balls to differentiate them into definitive endoderm cells; culturing the obtained definitive endoderm cells to induce their differentiation into pancreatic precursor cells; culturing the obtained pancreatic precursor cells to induce their differentiation into pancreatic islet β cells. The invention makes the test process and results more stable by adjusting the components of the culture solution in each step, and the culture solution is simpler, lower in cost and easy to store; the difference between single test results is smaller and the output is higher. The resulting cell mass is also more uniform and the cell performance is better. The induction of differentiation from pancreatic precursor cells to islet β cells can basically be regarded as a one-step culture. On the premise of ensuring effective directional differentiation of cells, it saves a lot of culture time and medium.

Description

technical field [0001] The invention relates to the technical field of application of stem cell differentiation in medical treatment, in particular to a method for obtaining pancreatic precursor cells and pancreatic islet beta cells by differentiation of human pluripotent stem cells. Background technique [0002] Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by defective insulin secretion, impaired biological action, or both. Long-term hyperglycemia leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. [0003] There is significant genetic heterogeneity in both type 1 and type 2 diabetes. Diabetes has a familial tendency, and 1 / 4-1 / 2 patients have a family history of diabetes. Clinically, there are at least 60 genetic syndromes associated with diabetes. Type 1 diabetes has multiple DNA loci involved in the pathogenesis, among which the DQ locus polymo...

AI Technical Summary

Problems solved by technology

[0004] Pancreatic islet transplantation is a fundamental method for the treatment of diabetes that emerged in this century. However, in the early stage, islet isolation and transplantation were mainly used, combined with appropriate immunosuppressive regimens. This kind of regimen is extremely dependent on donors, so it cannot be applied on a large scale

In recent years, the application of stem cell differentiation in the medical field has gradually attracted people's attention and preliminary development. Some studies have also begun to focus on the preparation of islet cells through stem cell differentiation. However, the current research methods have problems such as poor reproducibility and low yield. application, and cannot be widely extended to clinical application