32201 20210121082901.0 doi 10.1371/journal.pone.0135830 sideral 92343 ART-2015-92343 eng Wright LS VSX2 and ASCL1 are indicators of neurogenic competence in human retinal progenitor cultures 2015 Access copy available to the general public Unrestricted Three dimensional (3D) culture techniques are frequently used for CNS tissue modeling and organoid production, including generation of retina-like tissues. A proposed advantage of these 3D systems is their potential to more closely approximate in vivo cellular microenvironments, which could translate into improved manufacture and/or maintenance of neuronal populations. Visual System Homeobox 2 (VSX2) labels all multipotent retinal progenitor cells (RPCs) and is known to play important roles in retinal development. In contrast, the proneural transcription factor Acheate scute-like 1 (ASCL1) is expressed transiently in a subset of RPCs, but is required for the production of most retinal neurons. Therefore, we asked whether the presence of VSX2 and ASCL1 could gauge neurogenic potential in 3D retinal cultures derived from human prenatal tissue or ES cells (hESCs). Short term prenatal 3D retinal cultures displayed multiple characteristics of human RPCs (hRPCs) found in situ, including robust expression of VSX2. Upon initiation of hRPC differentiation, there was a small increase in co-labeling of VSX2+ cells with ASCL1, along with a modest increase in the number of PKCa+ neurons. However, 3D prenatal retinal cultures lost expression of VSX2 and ASCL1 over time while concurrently becoming refractory to neuronal differentiation. Conversely, 3D optic vesicles derived from hESCs (hESC-OVs) maintained a robust VSX2+ hRPC population that could spontaneously co-express ASCL1 and generate photoreceptors and other retinal neurons for an extended period of time. These results show that VSX2 and ASCL1 can serve as markers for neurogenic potential in cultured hRPCs. Furthermore, unlike hESC-OVs, maintenance of 3D structure does not independently convey an advantage in the culture of prenatal hRPCs, further illustrating differences in the survival and differentiation requirements of hRPCs extracted from native tissue vs. those generated entirely in vitro. info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 3.057 2015 MULTIDISCIPLINARY SCIENCES 11 / 62 = 0.177 2015 Q1 T1 1.427 2015 Agricultural and Biological Sciences (miscellaneous) 2015 Q1 Medicine (miscellaneous) 2015 Q1 Biochemistry, Genetics and Molecular Biology (miscellaneous) 2015 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion (orcid)0000-0003-0349-9997 Pinilla I Universidad de Zaragoza Saha J Clermont JM Lien JS Borys KD Capowski EE Phillips MJ Gamm DM. 1004 646 Universidad de Zaragoza Dpto. Cirugía,Ginecol.Obstetr. Área Oftalmología 10, 8 (2015), 0135830 [22p.] PLoS One PloS one 1932-6203 1183206 http://zaguan.unizar.es/record/32201/files/texto_completo.pdf Versión publicada 109701 http://zaguan.unizar.es/record/32201/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:32201 articulos driver 2021-01-21-08:15:42 ARTICLE