Differentiation of human embryonic stem cells into functional cerebellar neurons for the treatment of cerebellar ataxia

Abstract

Event Abstract Back to Event Differentiation of human embryonic stem cells into functional cerebellar neurons for the treatment of cerebellar ataxia Slaven Erceg1*, Mohammad Ronaghi1, Victoria Moreno-Manzano1, Sergio Lainez1, Rosa Planells1, Ivan Zipancic1 and Miodrag Stojkovic1 1 Centro de Investigacion Principe Felipe (CIPF), Cellular Reprogramming Laboratory, Spain The cerebellum has been viewed as a structure that has critical roles in motor and sensory learning, motor coordination and vestibular ocular reflex. Many cerebellum related disorders require a cell therapy as a possible treatment of neural loss. The present study shows that the application of inductive signals involved in the early patterning in the cerebellar region in vivo (FGF and Wnt signalling and RA) followed by application of human BMP proteins (BMP4/6/7 and GDF7),human mitogens (SHH and JAG1)and human neurotrophins (BDNF and NT3),the factors involved in early GCP development,directs hESC differentiation into cerebellar neuron-like cells. The differentiated hESC showed granule cells phenotype expressing both markers of dorsal neurons (MATH1 and ZIC1) and markers specific for cerebellar granule cell markers (ZIPRO1,EN1,ZIC1,2,3, MEIS1,PDE1c,PAX6,TAG1,CYCLID2 and GABAá6R). In addition, cerebellar-like neurons expressed T-shaped polarity phenotype of cerebellar granule neurons. We also observed the expression of the most important markers for interneurons (PAX2) and Purkinje cells: LHX5, CALB1, PCP2 (L7) and GAD67. To our knowledge, this is the first study where we showed differentiation of hESC toward cerebellar-like cells and the functional activity of generated cells. The patch-clamp confirmed the specific cerebellar functional phenotype of these cells. In vivo implantation of differentiated hESC previously transfected with MATH-GFP construct, into the external granule cells layer of neonatal mice resulted in cell migration across molecular layer, Purkinje cell layer and settlement in the IGL layer which is supposed to be the final destination of mature granule cells. This is the first study showed that universal mechanisms involved in early development of cerebellum can be recapitulated in vitro using hESC.This protocol can be used to create in vivo strategy for cell replacement therapy using hESC to treat many cerebellum-related diseases. Conference: 41st European Brain and Behaviour Society Meeting, Rhodes Island, Greece, 13 Sep - 18 Sep, 2009. Presentation Type: Poster Presentation Topic: Poster presentations Citation: Erceg S, Ronaghi M, Moreno-Manzano V, Lainez S, Planells R, Zipancic I and Stojkovic M (2009). Differentiation of human embryonic stem cells into functional cerebellar neurons for the treatment of cerebellar ataxia. Conference Abstract: 41st European Brain and Behaviour Society Meeting. doi: 10.3389/conf.neuro.08.2009.09.139 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 09 Jun 2009; Published Online: 09 Jun 2009. * Correspondence: Slaven Erceg, Centro de Investigacion Principe Felipe (CIPF), Cellular Reprogramming Laboratory, Valencia, Spain, serceg@cipf.es Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Slaven Erceg Mohammad Ronaghi Victoria Moreno-Manzano Sergio Lainez Rosa Planells Ivan Zipancic Miodrag Stojkovic Google Slaven Erceg Mohammad Ronaghi Victoria Moreno-Manzano Sergio Lainez Rosa Planells Ivan Zipancic Miodrag Stojkovic Google Scholar Slaven Erceg Mohammad Ronaghi Victoria Moreno-Manzano Sergio Lainez Rosa Planells Ivan Zipancic Miodrag Stojkovic PubMed Slaven Erceg Mohammad Ronaghi Victoria Moreno-Manzano Sergio Lainez Rosa Planells Ivan Zipancic Miodrag Stojkovic Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.