Abstract
Background
An embryonic stem (ES) cell-derived eye-like structure, made up of neural retinal lineage cells, retinal pigment epithelial (RPE) cells, and lens cells was constructed in our laboratory. We have shown that cells from these eye-like structures can be integrated into the developing optic vesicle of chicks. The purpose of this study was to determine whether the cells from these eye-like structures can differentiate into retinal ganglion cells (RGCs) when transplanted into the vitreous of an injured adult mouse retina.
Methods
ES cells were induced to differentiate into eye-like structures in vitro for 6 or 11 days. Recipient mouse eyes were injected with NMDA to injure the RGCs prior to the transplantation. Sham-treated eyes received the same amount of carrier vehicle. Cells were extracted from the eye-like structures and transplanted into the vitreous of damaged and control eyes. The host eyes were analyzed both qualitatively and quantitatively by immunohistochemistry 10 days or 8 weeks after transplantation.
Results
Cells from the ES cell-derived eye-like structures were integrated into the RGC layer, and differentiated into neurons when transplanted into control (non-NMDA-treated) adult eyes. However, they rarely expressed RGC markers. When they were transplanted into NMDA-treated eyes, the cells spread on the surface of the retina and covered a relatively large area of the host RGC layer that had been injured by the NMDA. The cells from the ES cell-derived eye cells frequently differentiated into cells expressing RGC-specific markers, and formed a new RGC layer. In addition, a small number of these ES cell-derived cells were observed to extend axon-like processes toward the optic disc of the host. However, visually evoked responses could not be recorded from the visual cortex.
Discussion
These findings suggest that ES cell-derived eye-like structures contain cells that can differentiate into RG-like cells and regenerate a new RGC layer. These cells also appeared to be integrated into the retina and extend axon-like processes toward the optic nerve head.
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Acknowledgements
We thank Junichiro Sugimoto and Kyoko Takahashi for their excellent technical assistance and advice. We also thank Drs. Shin-ichi Hayashi and Hiromi Okuyama for establishing the EGFP-expressing D3 ES cell line, and Drs. Shin-ichi Nishikawa and Satomi Nishikawa for donating the PA6 cell line. We are also grateful to Drs. Shigeru Kinoshita, Hisato Kondo, Hisahiro Yoshida, and Masami Watanabe for their thoughtful advice. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.
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This study was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.We had full control of all primary data and we all agree to allow Graefe’s Archive for Clinical and Experimental Ophthalmology to review our data upon request.
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Aoki, H., Hara, A., Niwa, M. et al. Transplantation of cells from eye-like structures differentiated from embryonic stem cells in vitro and in vivo regeneration of retinal ganglion-like cells. Graefes Arch Clin Exp Ophthalmol 246, 255–265 (2008). https://doi.org/10.1007/s00417-007-0710-6
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DOI: https://doi.org/10.1007/s00417-007-0710-6