Concise Review: Human Pluripotent Stem Cells for the Modeling of Pancreatic β-Cell Pathology

Abstract

Pancreatic β‐cells are the only source of insulin. Disturbances in β‐cell development or function may thus result in insulin deficiency or excess, presenting as hyper‐ or hypoglycemia. It is increasingly evident that common forms of diabetes (types 1 and 2) are pathogenically heterogeneous. Development of efficient therapies is dependent on reliable disease models. Although animal models are remarkably useful research tools, they present limitations because of species differences. As an alternative, human pluripotent stem cell technologies offer multiple possibilities for the study of human diseases in vitro. In the last decade, advances in the derivation of induced pluripotent stem cells from diabetic patients, combined with β‐cell differentiation protocols, have resulted in the generation of useful disease models for diabetes. First disease models have been focusing on monogenic diabetes. The development of genome editing technologies, more advanced differentiation protocols and humanized mouse models based on transplanted cells have opened new horizons for the modeling of more complex forms of β‐cell dysfunction. We present here the incremental progress made in the modeling of diabetes using pluripotent stem cells. We discuss the current challenges and opportunities of these approaches to dissect β‐cell pathology and devise new pharmacological and cell replacement therapies. stem cells 2019;37:33–41

Reprogrammed Cells Display Distinct Proteomic Signatures Associated with Colony Morphology Variability

“…A typical characterization pipeline to ensure pluripotency includes expression of singular pluripotency markers (SOX2 and POU5F1), karyotype analysis, and the ability to form the three germ layers using teratoma assays or embryoid body formation [1]. Despite these quality controls, numerous studies have shown major line-to-line variations [2–5]. To improve the utility of hPSCs in regenerative medicine and to ensure high-quality clinical-grade cell products, we need a pipeline of robust quality control methods that can be automated to benchmark the cells and filter out reprogrammed cells of inferior quality.…”

Short-Term Protocols to Obtain Insulin-Producing Cells from Rat Adipose Tissue: Signaling Pathways and In Vivo Effect

“…Our previous studies with EX-4 and resveratrol influenced this choice [41,42]. It is also important to emphasize that we look for functional insulin-secreting cells in this work rather than a pancreatic β cell phenotype, which takes in account, in addition to insulin secretion, other features such as morphology or islet aggregation (for a review, see [43]). We observed that protocols II (supplemented with EX-4 and activin A) and III (supplemented with resveratrol) generated cells with higher affinity for DTZ stain (indirect labeling of insulin presence) and pro-insulin (evaluated by Western blotting).…”

Functional Genomics in Pancreatic β Cells: Recent Advances in Gene Deletion and Genome Editing Technologies for Diabetes Research

“…One successful example is the restoration of FANCA gene expression in haematopoietic stem cells ( 164 ). HDR efficiency is generally low (less than 2%) but, with CRISPR technology, it can be improved to 10-40%, depending on the target region ( 165 , 166 ). Several attempts have been made to improve HDR efficiency by incorporating silent CRISPR-Cas-blocking mutations ( 167 ), suppressing NHEJ key molecules such as KU70, KU80, or DNA ligase IV ( 168 , 169 ), modification of RAD18 ( 165 ), providing asymmetric donor DNA ( 170 ) and applying chemicals such as scr7 ( 169 ).…”