title: Reprogramming of HUVECs into induced pluripotent stem cells (HiPSCs), generation and characterization of HiPSC-derived neurons and astrocytes.
abstract: Neurodegenerative diseases are characterized by chronic and progressive structural or functional loss of neurons. Limitations related to the animal models of these human diseases have impeded the development of effective drugs. This emphasizes the need to establish disease models using human-derived cells. The discovery of induced pluripotent stem cell (iPSC) technology has provided novel opportunities in disease modeling, drug development, screening, and the potential for "patient-matched" cellular therapies in neurodegenerative diseases. In this study, with the objective of establishing reliable tools to study neurodegenerative diseases, we reprogrammed human umbilical vein endothelial cells (HUVECs) into iPSCs (HiPSCs). Using a novel and direct approach, HiPSCs were differentiated into cells of central nervous system (CNS) lineage, including neuronal, astrocyte and glial cells, with high efficiency. HiPSCs expressed embryonic genes such as nanog, sox2 and Oct-3/4, and formed embryoid bodies that expressed markers of the 3 germ layers. Expression of endothelial-specific genes was not detected in HiPSCs at RNA or protein levels. HiPSC-derived neurons possess similar morphology but significantly longer neurites compared to primary human fetal neurons. These stem cell-derived neurons are susceptible to inflammatory cell-mediated neuronal injury. HiPSC-derived neurons express various amino acids that are important for normal function in the CNS. They have functional receptors for a variety of neurotransmitters such as glutamate and acetylcholine. HiPSC-derived astrocytes respond to ATP and acetylcholine by elevating cytosolic Ca2+ concentrations. In summary, this study presents a novel technique to generate differentiated and functional HiPSC-derived neurons and astrocytes. These cells are appropriate tools for studying the development of the nervous system, the pathophysiology of various neurodegenerative diseases and the development of potential drugs for their treatments.
authors: Department of Medicine, University of Alberta, Edmonton, Canada.
authors: Yohannes
authors: Y
authors: Haile
authors: Department of Medicine, University of Alberta, Edmonton, Canada.
authors: Maryam
authors: M
authors: Nakhaei-Nejad
authors: Department of Pharmacology, University of Alberta, Edmonton, Canada; Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada.
authors: Paul A
authors: PA
authors: Boakye
authors: Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada; Department of Psychiatry (Neurochemical Research Unit), University of Alberta, Edmonton, Canada.
authors: Glen
authors: G
authors: Baker
authors: Department of Pharmacology, University of Alberta, Edmonton, Canada; Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada.
authors: Peter A
authors: PA
authors: Smith
authors: Department of Medicine, University of Alberta, Edmonton, Canada.
authors: Allan G
authors: AG
authors: Murray
authors: Department of Medicine, University of Alberta, Edmonton, Canada; Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada.
authors: Fabrizio
authors: F
authors: Giuliani
authors: Department of Medicine, University of Alberta, Edmonton, Canada.
authors: Nadia
authors: N
authors: Jahroudi
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000166:cytology*; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D009474:Neurons / Q000166:cytology*
chemical_list: D000255:Adenosine Triphosphate; D000109:Acetylcholine; D002118:Calcium