RT Journal Article A1 Zhu, Danhong A1 Hikita, Sherry T. A1 Deng, Xuemei A1 Lu, Bo A1 Johnson, Lincoln V. A1 Clegg, Dennis O. A1 Tai, Yuchong A1 Ahuja, Ashish A1 Humayun, Mark A1 Hinton, David R. T1 Culture of Polarized Embryonic Stem Cell-Derived RPE on Synthetic Substrates JF Investigative Ophthalmology & Visual Science JO Invest. Ophthalmol. Vis. Sci. YR 2011 VO 52 IS 14 SP 3191 OP 3191 SN 1552-5783 AB RPE cell loss in the macular region is a key pathologic alteration in atrophic age-related macular degeneration (AMD). The restoration of lost RPE through transplantation of a polarized RPE monolayer is a promising new treatment for AMD. RPE cells derived from human embryonic stem cells (hESC-RPE) are a potentially unlimited resource for this therapy. This study tested two synthetic substrates, parylene (non-degradable) and poly(lactic-co-glycolic acid) (PLGA) (bio-degradable) as supports for the growth of polarized hESC-RPE monolayers for potential use in sub-retinal transplantation. 1). hESC-RPE were derived from H9 human embryonic stem cells.2). hESC-RPE were seeded on CellStart-coated (Invitrogen) substrate membranes at 105 cells/cm2 and cultured in XVIVO-10medium (Lonza) for 4 weeks.3). Scanning and transmission electron microscopy were used for ultrastructural analysis of cells and substrates.4).Whole human genome microarray analysis (Agilent) was performed on a parallel culture of hESC-RPE cells, where the cells were seeded on CellStart-coated tissue culture plates and grown in XVIVO-10 medium. RNA was collected at 4 weeks post-seeding. hESC-RPE attached, proliferated and differentiated into polarized monolayers on both substrates. They exhibited hexagonal shape, melaninization, and apical microvilli. Microarray results showed that hESC-RPE cells grown in XVIVO10 media expressed signature RPE genes and had higher amounts of PEDF, bestrophin and RDH5, compared with controls. Both synthetic substrates supported hESC-RPE cell growth and polarization. They are candidate platforms for differentiation and surgical manipulation of hESC-RPE monolayers for sub-retinal transplantation. RD 3/5/2021