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The vertebrate retina is a highly multilayered nervous tissue with a large diversity of cellular components. With the development of stem cell technologies, human retinas can be generated in three-dimensional (3-D) culture  in vitro . However, understanding the factors modulating key productive processes and the way that they influence development are far from clear. Oxygen, as the most essential element participating in metabolism, is a critical factor regulating organic development. In this study, using 3-D culture of human stem cells, we examined the effect of intermittent high oxygen treatment (40% O 2 ) on the formation and cellular behavior of neural retinas (NR) in the embryonic body (EB). The volume of EB and number of proliferating cells increased significantly under 40% O 2  on day 38, 50, and 62. Additionally, the ratio of PAX6+ cells within NR was significantly increased. The neural rosettes could only develop with correct apical-basal polarity under 40% O 2 . In addition, the generation, migration and maturation of retinal ganglion cells were enhanced under 40% O 2 . All of these results illustrated that 40% O 2  strengthened the formation of NR in EB with characteristics similar to the  in vivo  state, suggesting that the hyperoxic state facilitated the retinal development  in vitro .

Intermittent high oxygen influences the formation of neural retinal tissue from human embryonic stem cells