Subretinal Implantation of Retinal Pigment Epithelial Cells Derived From Human Embryonic Stem Cells: Improved Survival When Implanted as a Monolayer

Subretinal Implantation of Retinal Pigment Epithelial Cells Derived From Human Embryonic Stem Cells: Improved Survival When Implanted as a Monolayer

Author Diniz, Bruno Autor UNIFESP Google Scholar
Thomas, Padmaja Google Scholar
Thomas, Biju Google Scholar
Ribeiro, Ramiro Autor UNIFESP Google Scholar
Hu, Yuntao Google Scholar
Brant, Rodrigo Autor UNIFESP Google Scholar
Ahuja, Ashish Google Scholar
Zhu, Danhong Google Scholar
Liu, Laura Google Scholar
Koss, Michael Google Scholar
Maia, Mauricio Autor UNIFESP Google Scholar
Chader, Gerald Google Scholar
Hinton, David R. Google Scholar
Humayun, Mark S. Google Scholar
Institution Doheny Eye Inst
Universidade Federal de São Paulo (UNIFESP)
Univ So Calif
Hosp Univ Curitiba
Peking Univ
Chang Gung Mem Hosp
Goethe Univ Frankfurt
Abstract PURPOSE. To evaluate cell survival and tumorigenicity of human embryonic stem cell-derived retinal pigment epithelium (hESC-RPE) transplantation in immunocompromised nude rats. Cells were transplanted as a cell suspension (CS) or as a polarized monolayer plated on a parylene membrane (PM).METHODS. Sixty-nine rats (38 male, 31 female) were surgically implanted with CS (n = 33) or PM (n = 36). Cohort subsets were killed at 1, 6, and 12 months after surgery. Both ocular tissues and systemic organs (brain, liver, kidneys, spleen, heart, and lungs) were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned. Every fifth section was stained with hematoxylin and eosin and analyzed histologically. Adjacent sections were processed for immunohistochemical analysis (as needed) using the following antibodies: anti-RPE65 (RPE-specific marker), anti-TRA-1-85 (human cell marker), anti-Ki67 (proliferation marker), anti-CD68 (macrophage), and anti-cytokeratin (epithelial marker).RESULTS. the implanted cells were immunopositive for the RPE65 and TRA-1-85. Cell survival (P = 0.006) and the presence of a monolayer (P < 0.001) of hESC-RPE were significantly higher in eyes that received the PM. Gross morphological and histological analysis of the eye and the systemic organs after the surgery revealed no evidence of tumor or ectopic tissue formation in either group.CONCLUSIONS. hESC-RPE can survive for at least 12 months in an immunocompromised animal model. Polarized monolayers of hESC-RPE show improved survival compared to cell suspensions. the lack of teratoma or any ectopic tissue formation in the implanted rats bodes well for similar results with respect to safety in human subjects.
Keywords age-related macular degeneration
retinal pigment epithelium
human embryonic stem cells
Language English
Sponsor California Institute of Regenerative Medicine
Research to Prevent Blindness
National Eye Institute
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Grant number California Institute of Regenerative Medicine: DR1-01444
National Eye Institute: EY03040
CAPES: BEX 2326-11-6
Date 2013-07-01
Published in Investigative Ophthalmology & Visual Science. Rockville: Assoc Research Vision Ophthalmology Inc, v. 54, n. 7, p. 5087-5096, 2013.
ISSN 0146-0404 (Sherpa/Romeo, impact factor)
Publisher Assoc Research Vision Ophthalmology Inc
Extent 5087-5096
Origin http://dx.doi.org/10.1167/iovs.12-11239
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000322637000085
URI http://repositorio.unifesp.br/handle/11600/36455

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