干细胞
胚胎干细胞
生物
细胞生物学
视网膜
视网膜
分子生物学
干细胞标记物
染色
复原素
细胞分化
视紫红质
生物化学
遗传学
基因
神经科学
作者
Joo Young Shin,D. Ma,Mi Sun Lim,Myung Soo Cho,Young Joo Kim,Hyeong Gon Yu
摘要
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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