Clin Exp Reprod Med.
2011 Dec;38(4):216-221. 10.5653/cerm.2011.38.4.216.
Subretinal transplantation of putative retinal pigment epithelial cells derived from human embryonic stem cells in rat retinal degeneration model
- Affiliations
-
- 1Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea. hgonyu@snu.ac.kr
- 2Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea.
- 3Research and Development Center, Jeil Pharmaceutical Co., Ltd., Seoul, Korea.
- 4Department of Ophthalmology, Seoul Paik Hospital, Inje University, Seoul, Korea.
- 5Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea.
- KMID: 1743678
- DOI: http://doi.org/10.5653/cerm.2011.38.4.216
Abstract
OBJECTIVE
To differentiate the human embryonic stem cells (hESCs) into the retinal pigment epithelium (RPE) in the defined culture condition and determine its therapeutic potential for the treatment of retinal degenerative diseases.
METHODS
The embryoid bodies were formed from hESCs and attached on the matrigel coated culture dishes. The neural structures consisting neural precursors were selected and expanded to form rosette structures. The mechanically isolated neural rosettes were differentiated into pigmented cells in the media comprised of N2 and B27. Expression profiles of markers related to RPE development were analyzed by reverse transcription-polymerase chain reaction and immunostaining. Dissociated putative RPE cells (10(5) cells/5 microL) were transplanted into the subretinal space of rat retinal degeneration model induced by intravenous sodium iodate injection. Animals were sacrificed at 1, 2, and 4 weeks after transplantation, and immnohistochemistry study was performed to verify the survival of the transplanted cells.
RESULTS
The putative RPE cells derived from hESC showed characteristics of the human RPE cells morphologically and expressed molecular markers and associated with RPE fate. Grafted RPE cells were found to survive in the subretinal space up to 4 weeks after transplantation, and the expression of RPE markers was confirmed with immunohistochemistry.
CONCLUSION
Transplanted RPE cells derived from hESC in the defined culture condition successfully survived and migrated within subretinal space of rat retinal degeneration model. These results support the feasibility of the hESC derived RPE cells for cell-based therapies for retinal degenerative disease.
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