Abstract
BackgroundHuman embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cell transplants have served as a cell therapy for treating retinal degenerative diseases. However, how to optimize the survival and engraftment of hESC-RPE cells is a great challenge.MethodsHere, we report hESC-RPE cells that are embedded with polyelectrolytes gelatin and alginate by layer-by-layer (LbL) self-assembly technique, based on the opposite charge of alternate layers. Cells were assessed for cell survival, immunogenicity, and function in vitro and in vivo.ResultsThis strategy obviously decreased the immunogenicity of hESC-RPE cells without affecting its activity. LbL-RPE cell transplants into the subretinal space of Royal College of Surgeons (RCS) rats optimized cell engraftment and decreased immunogenicity compared to untreated RPE cell transplants (immunosuppression was not used during the 21-week study). Visual-functional assay with electroretinogram recordings (ERGs) also showed higher B wave amplitudes in RCS rats with LbL-RPE cell transplants.ConclusionsWe demonstrate that transplanted LbL-RPE cells have better viability and grafting efficiency, optimized immunogenicity, and visual function. Therefore, LbL engineering is a promising method to increase the efficacy of hESC-RPE cell transplantation.
Highlights
Human embryonic stem cell-derived retinal pigment epithelial cell transplants have served as a cell therapy for treating retinal degenerative diseases
The culture medium for the Retinal pigment epithelial (RPE) cells that diffused from the excised pigment foci contained 78% KO-DMEM CTS (Invitrogen), 20% Knock Out SR xenofree CTS (Invitrogen), 1% CTS glutaMAX-1 supplement (Invitrogen), 1% MEM NEAA (Invitrogen), and 1% 2-Mercaptoethanol (Procell). hESC-derived RPE cells were cultured in cell culture dishes at 37 °C in an incubator with 5% CO2/95% air, and the medium replaced every 2 days
Preparation of layer-by-layer encapsulation of RPE cells The differentiation of hESC into RPE cells was summarized in Figure S1A, which includes three steps: superconfluence, acquired pigment foci, and excision
Summary
Human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cell transplants have served as a cell therapy for treating retinal degenerative diseases. How to optimize the survival and engraftment of hESC-RPE cells is a great challenge. One is the injection of an RPE cell suspension, and the other involves engrafting a monolayer of RPE cells seeded on a supporting membrane. For the former method, how to ensure that cells are engrafted to the targeted lesion area remains a serious challenge [3, 12, 13]. We successfully transplanted hESC-RPE suspension [14] into the subretinal space of AMD patients [10]. It is difficult to control the cells distribution after surgery
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