Abstract
Purpose :
Recently, the reports about the direct reprogramming of brain astrocytes into neural cells with administration of small-molecule chemical modulators increase gradually. However, anyone has not established the method of retinal neurogenesis from astrocytes. We thus investigated whether astrocytes could differentiate into retinal neural cells only with small-molecule chemical modulators.
Methods :
In vivo, we used the experimental mouse model of retinal detachment (RD) induced by subretinal injection of sodium hyaluronate, and performed the immunostaining of Nestin (the marker of neural progenitor cells). In vitro, we stimulated astrocytes with four small-molecule compounds. After these chemical stimulations, we examined the changes in thirty markers of neural progenitor and retinal progenitor cells using flow cytometry and real-time polymerase chain reaction (PCR) at several time-points.
Results :
In vivo, retinal astrocytes clearly expressed Nestin in RD mice, but these cells were not able to proliferate. In vitro, cultured astrocytes transformed into small round cells within one day after the stimulation with chemical modifiers, and their CD44 expression (marker of mature astrocytes) reduced significantly (P <0.05). Only a part of these round cells obtained the self-renewal ability, and proliferated as spheres after 2 weeks from the initial stimulation. The protein and gene expressions of Nestin, CD133, SOX2, Musashi-1 (the marker of neural progenitor cells) and CD15 (the marker of retinal progenitor cells) increased in these cells.
Conclusions :
We were able to induce mature astrocytes into neural progenitor cells only with small-molecule chemical modulators. These results suggested the possibility of retinal neural regeneration with administration of small-molecule compounds.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.