Influence of oxygen tension on the differentiation of embryonic stem cells towards photoreceptors and other retinal phenotypes

A thesis submitted for the degree of Philosophiae Doctor.Retinitis pigmentosa (RP), a genetically heterogeneous group of diseases together with age-related macular degeneration (AMD), are the leading causes of permanent blindness and are characterized by the progressive dysfunction and death of the light sensing photoreceptors of the retina. Due to the limited regeneration capacity of the mammalian retina the scientific community has invested significantly in trying to obtain retinal progenitor cells from embryonic stem cells (ESC). These represent an unlimited source of retinal cells, but it has not yet been possible to achieve specific populations, such as photoreceptors, efficiently enough to allow them to be used safely in the future as cell therapy of RP or AMD. In this study we generated a high yield of photoreceptors from directed differentiation of mouse ESC (mESC) by recapitulating crucial phases of retinal development. We present a new protocol of differentiation, involving hypoxia and taking into account extrinsic and intrinsic cues. These include niche-specific conditions as well as the manipulation of the signaling pathways involved in retinal development.Our results show that hypoxia promotes and improves the differentiation of mESC towards photoreceptors. Different populations of retinal cells are increased in number under the hypoxic conditions applied, such as Crx positive cells, S-Opsin positive cells and double positive cells for Rhodopsin and Recoverin, as shown by immunofluorescence analysis. For the first time we report the high efficiency of differentiation in vivo and the expression of mature rod photoreceptor markers in a large number of differentiated cells, transplanted in the sub-retinal space of wild type mice.The present project was possible thanks to the Fundacion Publica Andaluza Progreso y Salud, from Junta de Andalusia.Peer reviewe

Propuesta de diseño del edificio de subespecialidades médicas del complejo hospitalario Dr. Rafael Angel Calderón Guardia

Proyecto de graduación (licenciatura en arquitectura)UCR::Vicerrectoría de Docencia::Ingeniería::Facultad de Ingeniería::Escuela de Arquitectur

LA EDUCACIÓN SUPERIOR Y EL TRABAJO CON GRUPOS SOCIALES DE ATENCIÓN PRIORITARIA. THE HIGHER EDUCATION AND THE WORK WITH

Muchas comunidades urbanas de Costa Rica enfrentan problemáticas de desigualdad y vulnerabilidad social, como es el caso de Guararí, Los Lagos y La Aurora de Heredia. Las universidades públicas han unido esfuerzos, por medio de la Extensión Universitaria y Acción Social y con el financiamiento del Consejo Nacional de Rectores, para atender problemas reales de esas comunidades. Entre el 2009 y el 2010, se han desarrollado procesos formativos y dialógicos, con el fin de incidir en los modos de vida y en el aprovechamiento de oportunidades socio-económicas, individuales y colectivas. Este artículo presenta las experiencias obtenidas con la realización de un proyecto en particular, que contó con la participación de más de trescientas personas de la zona. Se realizaron acciones que van desde el  diagnóstico participativo hasta la ejecución de una serie de módulos de formación, de carácter no formal, que comprenden temas como la sexualidad, los estilos de vida saludables, las prácticas democráticas, el emprendedurismo, planes de negocios, entre otros.Palabras claves: Extensión Universitaria, docencia, emprendedurismo, desarrollo humano. Abstract In Costa Rica urban communities such as Guararí, Los Lagos and La Aurora in the province of Heredia face inequality and social vulnerability. In order to work towards the wellbeing of those communities public universities have joined forces to, through the University Extension and Social Action, address that reality.  With the funding of the National Council of Rectors (CONARE) the Universidad Nacional (UNA), the Universidad de Costa Rica (UCR) and the Universidad Estatal a Distancia (UNED) developed through 2009 -2010 a capacity building process with young people in order to influence their lifestyles and to facilitate socio-economic opportunities. This article presents the experiences gained through this project as well as the participation of more than three hundred people in those areas in actions ranging from the participatory assessment to implementation of a series of training non-formal modules, covering topics such as sexuality, healthy lifestyles, democratic practices, entrepreneurship, business plans, among others.Keywords: University extension, education, entrepreneurship, human development

La educación superior y el trabajo con grupos sociales de atención prioritaria

Muchas comunidades urbanas de Costa Rica enfrentan problemáticas de desigualdad y vulnerabilidad social, como es el caso de Guararí, Los Lagos y La Aurora de Heredia. Las universidades públicas han unido esfuerzos, por medio de la Extensión Universitaria y Acción Social y con el financiamiento del Consejo Nacional de Rectores, para atender problemas reales de esas comunidades. Entre el 2009 y el 2010, se han desarrollado procesos formativos y dialógicos, con el fin de incidir en los modos de vida y en el aprovechamiento de oportunidades socio-económicas, individuales y colectivas. Este artículo presenta las experiencias obtenidas con la realización de un proyecto en particular, que contó con la participación de más de trescientas personas de la zona. Se realizaron acciones que van desde el diagnóstico participativo hasta la ejecución de una serie de módulos de formación, de carácter no formal, que comprenden temas como la sexualidad, los estilos de vida saludables, las prácticas democráticas, el emprendedurismo, planes de negocios, entre otros.In Costa Rica urban communities such as Guararí, Los Lagos and La Aurora in the province of Heredia face inequality and social vulnerability. In order to work towards the wellbeing of those communities public universities have joined forces to, through the University Extension and Social Action, address that reality. With the funding of the National Council of Rectors (CONARE) the Universidad Nacional (UNA), the Universidad de Costa Rica (UCR) and the Universidad Estatal a Distancia (UNED) developed through 2009 -2010 a capacity building process with young people in order to influence their lifestyles and to facilitate socio-economic opportunities. This article presents the experiences gained through this project as well as the participation of more than three hundred people in those areas in actions ranging from the participatory assessment to implementation of a series of training non-formal modules, covering topics such as sexuality, healthy lifestyles, democratic practices, entrepreneurship, business plans, among others

Concise Review: Stem Cells for the Treatment of Cerebellar-Related Disorders

Embryonic neural transplants have become clinically relevant over the past 25 years for their possible application in the treatment of cerebellum-related neurodegenerative diseases. While highlighting the important role that fetal neural progenitors have in meeting these challenges, we define rationales for all types of cell therapy involving adult stem cells as well as human embryonic stem cells (hESC) and human induced pluripotent stem (iPS) cells. The recent advances in the field of hESC and iPS cells, including their capacity for differentiation toward regional specific neural lineages, could open a new era of transplantation in cell-based therapy for cerebellar ataxiasFunded by “Miguel Servet” contract of Instituto de Salud Carlos III of Spanish Ministry of Science and Innovation Fund for Health of Spain. Grant Number: PI10-01683 Junta de Andalucia. Grant Number: PI-0113-2010Peer Reviewe

Hypoxia increases the yield of photoreceptors differentiating from mouse embryonic stem cells and improves the modeling of retinogenesis in vitro.

Retinitis pigmentosa (RP), a genetically heterogeneous group of diseases together with age-related macular degeneration (AMD), are the leading causes of permanent blindness and are characterized by the progressive dysfunction and death of the light sensing photoreceptors of the retina. Due to the limited regeneration capacity of the mammalian retina, the scientific community has invested significantly in trying to obtain retinal progenitor cells from embryonic stem cells (ESC). These represent an unlimited source of retinal cells, but it has not yet been possible to achieve specific populations, such as photoreceptors, efficiently enough to allow them to be used safely in the future as cell therapy of RP or AMD. In this study, we generated a high yield of photoreceptors from directed differentiation of mouse ESC (mESC) by recapitulating crucial phases of retinal development. We present a new protocol of differentiation, involving hypoxia and taking into account extrinsic and intrinsic cues. These include niche-specific conditions as well as the manipulation of the signaling pathways involved in retinal development. Our results show that hypoxia promotes and improves the differentiation of mESC toward photoreceptors. Different populations of retinal cells are increased in number under the hypoxic conditions applied, such as Crx-positive cells, S-Opsin-positive cells, and double positive cells for Rhodopsin and Recoverin, as shown by immunofluorescence analysis. For the first time, this manuscript reports the high efficiency of differentiation in vivo and the expression of mature rod photoreceptor markers in a large number of differentiated cells, transplanted in the subretinal space of wild-type mice. STEM CELLS 2013;31:966¿978This work was supported by funds for research from Junta de Andalucía PI-0113-2010 (S.E.) and ‘‘Miguel Servet’’ contract of Instituto de Salud Carlos III of Spanish Ministry of Science and Innovation (S.E.).Peer Reviewe