Isolation and transplantation of retinal stem cells

Abstract

The present invention relates to the isolation, in vitro propagation, and transplantation and integration of non-pigmented retinal stem cells derived from the neuroretina of the eye, ex vivo and in vivo.

1 . Retinal stem cells isolated from a neuroretina of a donor mammal. 
   
   
       2 . Retinal stem cells isolated from a neuroretina of a donor mammal, being capable of:
 a) self-renewal in vitro;   b) differentiating into any one cell type of the group consisting of neurons, astrocytes, and oligodendrocytes;   c) integration into a host retina when grafted on the host retina; and   d) differentiation into photoreceptor cells when grafted onto a retina, onto a retinal explants, or into a mature eye.   
   
   
       3 . The retinal stem cells of  claim 1  or  2 , wherein the retinal stem cells express nestin and are non-pigmented. 
   
   
       4 . The retinal stem cells of  claim 1  or  2 , said cells requiring, when cultured in vitro, the presence at least one exogenous growth factor in a culture medium in order to proliferate. 
   
   
       5 . The retinal stem cells of  claim 4 , wherein at least one exogenous growth factor is a member selected from the group consisting of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), a combination of bFGF and EGF, and a combination of EGF and bFGF and platelet-derived growth factor (PDGF). 
   
   
       6 . The retinal stem cells of  claim 1  or  2 , which can differentiate into neurons, as evidenced by expression of a neurofilament protein, NF-200. 
   
   
       7 . The retinal stem cells of  claim 1  or  2 , which can differentiate into astrocytes, as evidenced by expression of glial fibrillary acidic protein, GFAP. 
   
   
       8 . The retinal stem cells of  claim 1  or  2 , which, when grafted onto a retina, onto a retinal explants, or into a mature eye, differentiate into photoreceptor cells expressing rhodopsin, recoverin, or both. 
   
   
       9 . The retinal stem cells of  claim 8 , further being capable of integrating into and repopulating a diseased retina. 
   
   
       10 . The retinal stem cells of  claim 1  or  2 , wherein the mammal is an embryo, a neonate, or an adult. 
   
   
       11 . The retinal stem cells of  claim 1  or  2 , wherein the mammal is a primate, a rodent, or a domesticated animal. 
   
   
       12 . The retinal stem cells of  claim 1  or  2 , wherein the mammal is a human, a mouse, a rat, a cat, a dog, a pig, a cow, a horse, a monkey, or a great ape. 
   
   
       13 - 24 . (canceled) 
   
   
       25 . The neuroretina-derived retinal stem cells of  claim 1  or  2 , wherein the mammal was is a transgenic mouse expressing green fluorescent protein. 
   
   
       26 . The neuroretina-derived retinal stem cells of  claim 1  or  2 , being clonally derived. 
   
   
       27 - 45 . (canceled) 
   
   
       46 . The retinal stem cells of  claim 1  which are derived from a neurosphere.