Published in: First Personal Medical Congress
COI code: IPMCMED01_135
Paper Language: English
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Authors A Novel iPSC Model of Parkinson’s Disease Based on IFNB-Knockout MiceMojtaba Nemati - Royan Institute
Adele Taee - Royan Institute
Mahdi Hesaraki - Royan Institute
Shohreh Issazadeh-Navikas - University of Copenhagen
Abstract:Background:Parkinson’s disease is an elusive condition causing the loss of dopaminergic neurons in the Substantia Nigra with not much being known about its underlying mechanisms. Despite many efforts to model this disease both in vivo and in vitro, until recently none have been completely successful. We have used cells from the recently developed mouse model based on the knockout of Interferon beta to create induced pluripotent stem cells (iPSCs) which were then differentiated to dopaminergic neurons. After characterizing their Parkinsonian phenotype a novel in vitro model for Parkinson’s disease was developed. Materials and Methods:iPSCs were generated from both KO and WT mouse-fibroblasts using retroviral induction of the four Yamanaka factors Oct4, Sox2, KLF4 and C-Myc. The generated iPSCs were characterized using Alkaline phosphatase assay as well as immunofluorescence and qRT-PCR to confirm the expression for pluripotency factors. After characterization these cells were differentiated to dopaminergic neurons using the dual SMAD inhibition method of neural induction. Complete differentiation to functional dopaminergic neurons were assessed using immunofluorescence for Tyrosine Hydroxylase, and GIRK2 and electrophysiological tests (patch clamp). Immunofluorescence imaging using an antibody against phosphorylated Alpha-Synuclein was used to check for Lewy bodies.Results:Our results show the aggregation of phosphorylated alpha-synuclein which is the hallmark of Parkinson’s disease in the the KO neurons, as detected using immunofluorescence, but not in the neurons differentiated from WT iPSCs.Conclusion:This model is the only model to date which exhibits Lewy body formation in vitro, we have an unprecedented opportunity to study the mechanisms of Parkinson’s disease The setup of this procedure allows individuals identified as being at risk of this disease to be tested using in vitro modeling. If they are found to be susceptible, drug screening can be used to tailor a personalized treatment.
Keywords:Parkinson’s Disease, IPS model, Disease Modeling, Interferon Beta
COI code: IPMCMED01_135
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Nemati, Mojtaba; Adele Taee; Mahdi Hesaraki & Shohreh Issazadeh-Navikas, 2016, A Novel iPSC Model of Parkinson’s Disease Based on IFNB-Knockout Mice, First Personal Medical Congress, تهران, دانشگاه علوم پزشكي ايران - پژوهشكده ملي مهندسي ژنتيك و زيست فناوري ايران، مركز همكاري هاي فناوري و نوآوري هاي رياست جمهوري, https://www.civilica.com/Paper-IPMCMED01-IPMCMED01_135.htmlInside the text, wherever referred to or an achievement of this article is mentioned, after mentioning the article, inside the parental, the following specifications are written.
First Time: (Nemati, Mojtaba; Adele Taee; Mahdi Hesaraki & Shohreh Issazadeh-Navikas, 2016)
Second and more: (Nemati; Taee; Hesaraki & Issazadeh-Navikas, 2016)
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Type: Research Center
Paper No.: 5254
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