Corneal Spheres derived from Human Embryonic and Human Pluripotent Parthenogenetic Stem Cells

Abstract

Corneal blindness is common. Cornea transplants are the most commonly performed organ transplants, but the need for corneal grafts worldwide far outweighs the supply of healthy donor corneas. Here we describe a differentiation protocol that yields corneal orbs from human embryonic stem cells (hESC) as well as from human pluripotent parthenogenetic stem cells (hpSC), and therefore can be manufactured free of transmissible pathogens. Cornea and other tissues generated from parthenogenetic stem cells that are homozygous at HLA loci have a distinct im-munologic advantage over fully allogeneic grafts, and this report is the first to describe multilayered cornea generated from hpSC. The differentiated corneal product is layered and anatomically similar to normal human cornea, expresses appropriate corneal markers at the mRNA and protein (and secreted protein) levels, and is permeable to topical ophthalmic drugs. This 3D stem cell-derived cornea is a foundational step in development of appropriately organized, functional corneal grafts from hESC and hpSC for use in in vitro assays as well as regenerative therapies.