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The final impact of all retinal dystrophies is the loss of photoreceptors; hence, there is a pressing need for research into replacement. Seminal work has shown that a simple three-dimensional culture system enables differentiation of human pluripotent stem cells to retinal organoids containing large numbers of photoreceptors developing alongside retinal neurons and M\u00fcller glia cells in a laminated structure that resembles the native retina. Despite these promising developments, current protocols show different efficiencies across pluripotent stem cells and result in retinal organoids with a mixture of photoreceptor cells at varying maturation states, along with nonphotoreceptor cell types. In this study, we investigated the impact of stage-specific addition of retinoic acid (RA), 9-cis-retinal, 11-cis-retinal, levodopa (l-DOPA), triiodothyronine (T3), and \u03b3-secretase inhibitor ((2S)-N-[(3,5-Difluorophenyl)acetyl]-l-alanyl-2-phenyl]glycine1,1-dimethylethyl ester2L [DAPT]) in the generation of cone and rod photoreceptors. Our results indicate that addition of RA\u2009+\u2009T3 during days 90 to 120 of differentiation enhanced the generation of rod and S-cone photoreceptor formation, while the combined addition of DAPT from days 28 to 42 with RA during days 30 to 120 of differentiation led to enhanced generation of L\/M-cones at the expense of rods. l-DOPA when added together with RA during days 90 to 120 of differentiation also promoted the emergence of S-cones at the expense of rod photoreceptors. Collectively, these data represent an advance in our ability to direct generation of rod and cone photoreceptors in vitro.<\/jats:p>\n                  <jats:sec>\n                    <jats:title\/>\n                  <\/jats:sec>","DOI":"10.1002\/stem.3082","type":"journal-article","created":{"date-parts":[[2019,10,31]],"date-time":"2019-10-31T07:51:00Z","timestamp":1572508260000},"page":"45-51","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":56,"title":["Developing a simple method to enhance the generation of cone and rod photoreceptors in pluripotent stem cell-derived retinal organoids"],"prefix":"10.1093","volume":"38","author":[{"given":"Darin","family":"Zerti","sequence":"first","affiliation":[{"name":"a Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birthe","family":"Dorgau","sequence":"additional","affiliation":[{"name":"a Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Majed","family":"Felemban","sequence":"additional","affiliation":[{"name":"a Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK"},{"name":"b Princess Al-Jawhara Center of Excellence in Research of Hereditary Disorders and Department of Medical Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali E.","family":"Ghareeb","sequence":"additional","affiliation":[{"name":"a Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Yu","sequence":"additional","affiliation":[{"name":"a Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuchun","family":"Ding","sequence":"additional","affiliation":[{"name":"c Interdisciplinary Computing and Complex Biosystems (ICOS) Research Group, Newcastle University, Newcastle upon Tyne, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalio","family":"Krasnogor","sequence":"additional","affiliation":[{"name":"c Interdisciplinary Computing and Complex Biosystems (ICOS) Research Group, Newcastle University, Newcastle upon Tyne, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1327-8573","authenticated-orcid":false,"given":"Majlinda","family":"Lako","sequence":"additional","affiliation":[{"name":"a Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, 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