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Embryonic stem cell-derived photoreceptor precursor cells differentiated by coculture with RPE cells.
Shin, Joo Young; Ma, DaeJoong; Lim, Mi-Sun; Cho, Myung Soo; Kim, Young Joo; Yu, Hyeong Gon.
Afiliación
  • Shin JY; Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea.
  • Ma D; Department of Ophthalmology, Seoul Metropolitan Government Boramae Hospital, Seoul, Republic of Korea.
  • Lim MS; Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea.
  • Cho MS; Department of Ophthalmology, Seoul National University Hospital, Seoul, Republic of Korea.
  • Kim YJ; R&D Center, Jeil Pharmaceutical Co., Ltd., Yongin-si, Republic of Korea.
  • Yu HG; Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Republic of Korea.
Mol Vis ; 27: 288-299, 2021.
Article en En | MEDLINE | ID: mdl-34012231
Purpose: To describe the derivation of photoreceptor precursor cells from human embryonic stem cells by coculture with RPE cells. Methods: Human embryonic stem cells were induced to differentiate into neural precursor cells and then cocultured with RPE cells to obtain cells showing retinal photoreceptor features. Immunofluorescent staining, reverse transcription-PCR (RT-PCR), and microarray analysis were performed to identify photoreceptor markers, and a cGMP assay was used for in vitro functional analysis. After subretinal injection in rat animal models, retinal function was determined with electroretinography and optokinetic response detection, and immunofluorescent staining was performed to assess the survival of the injected cells. Results: Cocultured cells were positive for rhodopsin, red and blue opsin, recoverin, and phosphodiesterase 6 beta on immunofluorescent staining and RT-PCR. Serial detection of stem cell-, neural precursor-, and photoreceptor-specific markers was noted in each stage of differentiation with microarray analysis. Increased cGMP hydrolysis in light-exposed conditions compared to that in dark conditions was observed. After the subretinal injection in the rats, preservation of optokinetic responses was noted up to 20 weeks, while electroretinographic response decreased. Survival of the injected cells was confirmed with positive immunofluorescence staining of human markers at 8 weeks. Conclusions: Cells showed photoreceptor-specific features when stem cell-derived neurogenic precursors were cocultured with RPE cells.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Fotorreceptoras / Células Madre / Epitelio Pigmentado de la Retina / Células Madre Embrionarias Humanas Límite: Humans Idioma: En Revista: Mol Vis Asunto de la revista: BIOLOGIA MOLECULAR / OFTALMOLOGIA Año: 2021 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Fotorreceptoras / Células Madre / Epitelio Pigmentado de la Retina / Células Madre Embrionarias Humanas Límite: Humans Idioma: En Revista: Mol Vis Asunto de la revista: BIOLOGIA MOLECULAR / OFTALMOLOGIA Año: 2021 Tipo del documento: Article Pais de publicación: Estados Unidos