Highly Efficient Differentiation of Human Pluripotent Stem Cells into Pancreatic Progenitors Co-expressing PDX1 and NKX6
Diabetes is a complex metabolic disorder, with no available treatment. Islet transplantation is currently practiced beta cell replacement therapy option, however, with major limitations. Human pluripotent stem cells (hPSCs) can be used as a scalable sourc
- PDF / 719,719 Bytes
- 13 Pages / 504.567 x 720 pts Page_size
- 86 Downloads / 226 Views
Highly Efficient Differentiation of Human Pluripotent Stem Cells into Pancreatic Progenitors Co-expressing PDX1 and NKX6.1 Bushra Memon and Essam M. Abdelalim Abstract Diabetes is a complex metabolic disorder, with no available treatment. Islet transplantation is currently practiced beta cell replacement therapy option, however, with major limitations. Human pluripotent stem cells (hPSCs) can be used as a scalable source for generation of insulin-secreting cells as hPSCs have high proliferative capacity and can differentiate into any tissue type. In vitro stepwise protocols have been designed for differentiating hPSCs into pancreatic lineages that finally give rise to beta cells; however, these hPSC-derived beta cells are dissimilar to adult human beta cells in key aspects of gene expression and functionality. Alternatively, pancreatic progenitors, when transplanted in the body, have been shown to mature into functional insulin-secreting beta cells, capable of reversing hyperglycemia. These pancreatic progenitors require the co-expression of PDX1, a transcription factor (TF) regulating pancreatic development, and NKX6.1, another TF key for beta cell maturation and function, to produce glucose-responsive beta cells. Given the crucial role played by NKX6.1, we optimized an in vitro differentiation protocol to enhance the generation of pancreatic progenitors co-expressing PDX1 and NKX6.1 by modulating cell density, matrix availability, and cellular dissociation. Key words hPSCs, Differentiation, Transcription factors, Beta cells, Protocol, Cell therapy
1
Introduction Diabetes is a common metabolic disorder prevalent throughout the world. Type I diabetes (T1D) is characterized by immunemediated destruction of beta cells, whereas type II diabetes (T2D) is characterized by insulin resistance, which ultimately results in beta cell dysfunction [1]. Exogenous insulin supplementation does not provide robust physiological control over glycemic levels like the human islets. Therefore, recent therapeutic options such as cell therapy focus on restoring the endogenous beta cell resource. Allogenic islet transplantation is the most widely practiced beta cell replacement option; however, it also presents several limitations such as availability of adequate and functional cadaveric islets for transplantation and the constant need for immunosuppressants [2, 3]. hPSCs, including human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs),
Bushra Memon and Essam M. Abdelalim
can provide unlimited supply of pancreatic beta cells in vitro. These cells have a great potential to be used as cell therapy treatments, as well as to create an in vitro model of the disease. In case of diabetes, pancreatic beta cells derived from patient-specific PSCs harboring the disease genotype can be cultured and used as a model to reestablish the pathogenesis in vitro (see reviews [4–6]). Sequential directed differentiation protocols have been designed by many groups for generation of hPSC-derived pancreatic lineage products [7–
Data Loading...
