Method for obtaining pancreatic precursor cells and pancreatic beta cells through human pluripotent stem cell differentiation

Abstract

The invention discloses a method for obtaining pancreatic precursor cells and pancreatic beta cells through human pluripotent stem cell differentiation, and relates to the technical field of application of stem cell differentiation in medical treatment, and the method comprises the following steps: preparing human pluripotent stem cells into multifunctional cell balls; culturing the obtained multifunctional cell balls, and directionally differentiating the multifunctional cell balls into fixed endoderm cells; culturing the obtained fixed endoderm cells, and inducing the fixed endoderm cells to be differentiated into pancreatic precursor cells; and culturing the obtained pancreatic precursor cells and inducing the pancreatic precursor cells to be differentiated into pancreatic beta cells. By adjusting the components of the culture solution in each step, the test process and result are more stable, and the culture solution is simpler, low in cost and easy to store; the difference between single test results is smaller, and the yield is higher; and the obtained cells are more uniform in quality and better in cell performance. One-step culture can be basically considered from induced differentiation of pancreatic precursor cells into pancreatic beta cells, and a large amount of culture time and culture medium are saved on the premise that effective directional differentiation of the cells is guaranteed.

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 or impaired biological action, or both. Long-term high blood sugar leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. [0003] Significant genetic heterogeneity exists in both type 1 and type 2 diabetes. Diabetes tends to occur in families, and 1 / 4-1 / 2 patients have a family history of diabetes. Clinically, there are at least 60 genetic syndromes that can be associated with diabetes. Multiple DNA sites are involved in the pathogenesis of type 1 diabetes, among whi...

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