Human pluripotent stem (hPS) cells can differentiate into any cell type in the human body, including pancreatic islet cells. The current methods used to differentiate hPS cells into mature, insulin-producing beta cells are not efficient or consistent enough to be used for diabetes cell therapies. In this new study, published in 'Scientific Reports', researchers from the Paul Langerhans Institute Dresden (PLID) of the German Center for Diabetes Research, have developed hPS cell fluorescent reporter lines that can be used to monitor the progress and efficiency of the differentiation into functional pancreatic islet cells.

Pancreatic beta cells produce insulin, the hormone that regulates the uptake of glucose from the blood into the cells of the body. The loss or impaired function of these cells leads to diabetes, a disease in which the body cannot produce sufficient amounts of insulin. Current treatments, particularly for type 1 diabetes, include insulin injections or pancreas transplants, but these therapies have their limitations. Another possible future diabetes therapy would be the use of pancreatic islet cells, including insulin-producing beta cells derived from hPS cells. The use of hPS cells for diabetes cell therapies would be able to overcome the limitations of current treatments. However, the process of differentiation of hPS cells into beta cells is currently not well enough understood. Furthermore, it will be necessary to improve the current differentiation methods for the production of mature, insulin-producing cells.

To follow the differentiation process, the researchers in this study used CRISPR-Cas9 gene editing. “We created hPS cell reporter lines in which fluorescent proteins reflect the expression of key islet-specific proteins such as the hormones insulin (INS) and glucagon (GCG) and the transcription factor MAFA,” explains Prof. Anthony Gavalas, lead author of the study. “Visualizing these proteins allowed us to follow the development and maturation of different islet cell types in real time, giving us a detailed insight into the differentiation process.” The researchers used these lines and created an automated high-throughput imaging pipeline that helped to identify a new additive called N21, which is able to increase the number of insulin-producing cells.

 

© Anthony Gavalas | PLID
 

The ability to reliably produce functional beta cells from hPS could reduce reliance on insulin injections or pancreas transplants, providing a more sustainable and effective treatment option for people with type 1 diabetes. “The hPS cell reporter lines and the high-content live imaging approach described in this study allow efficient assessment of the various conditions for optimal islet cell differentiation and maturation, including functional beta cells,” concludes Prof. Gavalas. The use of advanced genetic engineering techniques and real-time imaging thus paves a new path for future innovations in the treatment of diabetes and related diseases.

Original publication:
Elisa Zanfrini, Manuj Bandral, Luka Jarc, Maria Alejandra Ramirez-Torres, DanielaPezzolla, Vida Kufrin, Eva Rodriguez-Aznar, Ana Karen Mojica Avila, Christian Cohrs, Stephan Speier, Katrin Neumann, Anthony Gavalas (2024). Generation and application of novel hES cell reporter lines for the differentiation and maturation of hPS cell-derived islet-like clusters. Sci Rep 14, 19863 (2024). doi.org/10.1038/s41598-024-69645-4

 

The Technische Universität Dresden (TU Dresden) is one of Germany's niversities of Excellence, esteemed for its exceptional standards in research and teaching spanning diverse fields. The Faculty of Medicine at TU Dresden is dedicated to propelling medical science and healthcare forward through interdisciplinary collaboration and pioneering research. https://www.uniklinikum-dresden.de/en

The Paul Langerhans Institute of Helmholtz Munich at the University Hospital Carl Gustav Carus and the Faculty of Medicine at TU Dresden (PLID) contributes decisively to a better understanding of the mechanisms of the disease and to explore new therapeutic options. The institute is a founding-partner of the German Center for Diabetes Research (DZD e.V.) and has been a satellite institute of Helmholtz Munich since January 2015. Its program comprises research into the pathophysiology of type 1 and type 2 diabetes mellitus. The main focus is on the mechanisms which cause the destruction and/or limited function of pancreatic beta cells and insufficient insulin secretion. In addition, the PLID also plays an outstanding role as only German transplant center for human pancreatic islet cells. https://tu-dresden.de/med/mf/plid

The German Center for Diabetes Research (DZD) is a national association that brings together experts in the field of diabetes research and combines basic research, translational research, epidemiology and clinical applications. The aim is to develop novel strategies for personalized prevention and treatment of diabetes. Members are Helmholtz Munich – German Research Center for Environmental Health, the German Diabetes Center in Düsseldorf, the German Institute of Human Nutrition in Potsdam-Rehbrücke, the Paul Langerhans Institute Dresden of Helmholtz Munich at the University Medical Center Carl Gustav Carus of the TU Dresden and the Institute for Diabetes Research and Metabolic Diseases of Helmholtz Munich at the Eberhard-Karls-University of Tuebingen together with associated partners at the Universities in Heidelberg, Cologne, Leipzig, Lübeck and Munich. www.dzd-ev.de/en