A novel approach for subretinal implantation of ultrathin substrates containing stem cell-derived retinal pigment epithelium monolayer

TitleA novel approach for subretinal implantation of ultrathin substrates containing stem cell-derived retinal pigment epithelium monolayer
Publication TypeJournal Article
Year of Publication2012
AuthorsHu Y, Liu L, Lu B, Zhu D, Ribeiro R, Diniz B, Thomas PB, Ahuja AK, Hinton DR, Tai Y-C, Hikita ST, Johnson LV, Clegg DO, Thomas BB, Humayun MS
JournalOphthalmic Research
Volume48
Issue4
Pagination186-91
Date Published2012
ISSN1423-0259
KeywordsAnimals, Cell Count, Embryonic Stem Cells, Feasibility Studies, Humans, Polymers, Rats, Rats, Mutant Strains, Retina, Retinal Dystrophies, Retinal Pigment Epithelium, Stem Cell Transplantation, Tissue Engineering, Tissue Scaffolds, Tomography, Optical Coherence, Xylenes
Abstract

OBJECTIVE: To evaluate the feasibility of a new technique for the implantation of ultrathin substrates containing stem cell-derived retinal pigment epithelium (RPE) cells into the subretinal space of retina-degenerate Royal College of Surgeon (RCS) rats.

METHODS: A platform device was used for the implantation of 4-µm-thick parylene substrates containing a monolayer of human embryonic stem cell-derived RPE (hESC-RPE). Normal Copenhagen rats (n = 6) and RCS rats (n = 5) were used for the study. Spectral-domain optical coherence tomography (SD-OCT) scanning and histological examinations were performed to confirm placement location of the implant. hESC-RPE cells attached to the substrate before and after implantation were evaluated using standard cell counting techniques.

RESULTS: SD-OCT scanning and histological examination revealed that the substrates were precisely placed in the rat's subretinal space. The hESC-RPE cell monolayer that covered the surface of the substrate was found to be intact after implantation. Cell counting data showed that less than 2% of cells were lost from the substrate due to the implantation procedure (preimplantation count 2,792 ± 74.09 cells versus postimplantation count 2,741 ± 62.08 cells). Detailed microscopic examination suggested that the cell loss occurred mostly along the edges of the implant.

CONCLUSION: With the help of this platform device, it is possible to implant ultrathin substrates containing an RPE monolayer into the rat's subretinal space. This technique can be a useful approach for stem cell-based tissue bioengineering techniques in retinal transplantation research.

DOI10.1159/000338749
Alternate JournalOphthalmic Res.
PubMed ID22868580
Grant ListEY03040 / EY / NEI NIH HHS / United States
P30 EY003040 / EY / NEI NIH HHS / United States