Retinal pigment epithelium cell compositions

Abstract

Presented herein are ready to administer (RTA) retinal pigment epithelium (RPE) cell therapy compositions for the treatment of retinal degenerative diseases and injuries. A method of formulating human RPE cells for administration to a subject directly after thawing and of formulating RPE cell therapy compositions for cryopreservation and administration of the cryopreserved composition to a subject subsequent to thawing are also presented. In another aspect, the RTA composition may be formulated as a thaw and inject (TAI) composition, whereby the composition is administered by injection subsequent to thawing.

1 - 62 . (canceled) 
     
     
         63 . A composition comprising:
 (a) adenosine, dextran-40, HEPES (N-(2-Hydroxyethyl) piperazine-N′-(2-ethanesulfonic acid)), L glutathione, and dimethyl sulfoxide (DMSO); and   (b) RPE cells, wherein the composition can be stored at cryothermic temperatures and wherein the composition is ready to administer to a subject directly after thawing.   
     
     
         64 - 65 . (canceled) 
     
     
         66 . The composition of  claim 63 , further comprising one or more of: a sugar acid, a base, an antioxidant, halide salt, a basic salt, phosphate salt, sugar, and sugar alcohol. 
     
     
         67 . The composition of  claim 66 , wherein the sugar acid comprises lactobionic acid, glyceric acid, xylonic acid, gluconic acid, ascorbic acid, neuraminic acid, ketodeoxyoctulosonic acid, glucuronic acid, galacturonic acid, galacturonic acid, iduronic acid, tartaric acid, mucic acid, and/or saccharic acid. 
     
     
         68 . The composition of  claim 66 , wherein the base comprises sodium hydroxide and/or potassium hydroxide. 
     
     
         69 . The composition of  claim 66 , wherein the antioxidant comprises L-glutathione, ascorbic acid, lipoic acid, uric acid, a carotene, alpha-tocopherol, and/or ubiquinol. 
     
     
         70 . The composition of  claim 66 , wherein the halide salt comprises potassium chloride, sodium chloride, and/or magnesium chloride. 
     
     
         71 . The composition of  claim 66 , wherein the basic salt comprises potassium bicarbonate, sodium bicarbonate, and/or sodium acetate. 
     
     
         72 . The composition of  claim 66 , wherein the phosphate salt comprises potassium phosphate, sodium phosphate, and/or potassium phosphate. 
     
     
         73 . The composition of  claim 66 , wherein the one or more sugars comprises dextrose and/or sucrose. 
     
     
         74 . The composition of  claim 66 , wherein the sugar alcohol comprises mannitol, sorbitol, erythritol and/or xylitol. 
     
     
         75 . (canceled) 
     
     
         76 . The composition of  claim 63 , wherein the RPE cell concentration is between about 100,000 and about 10,000,000 cells/ml. 
     
     
         77 . The composition of  claim 63 , wherein the number of cells in said composition is between about 100,000 to about 500,000. 
     
     
         78 . (canceled) 
     
     
         79 . The composition of  claim 63 , further comprising one or more of ROCK inhibitor or nicotiname (NA). 
     
     
         80 . The composition of  claim 63 , wherein the cells demonstrate one or more of the following after thawing: a barrier function TEER of about 100Ω to about 1300Ω; a PEDF Upper to Lower Ratio of about 3.5 to about 9.4; a VEGF Lower to Upper Ratio of about 1.2 to about 5; or a purity of about 95% to about 100%. 
     
     
         81 . The composition of  claim 63 , wherein the cells secrete one or more of-fibroblast growth factors (bFGF and aFGF), ciliary neurotrophic factor (CNTF), pigment epithelium-derived factor (PEDF), brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF) or one or more anti-inflammatory cytokines. 
     
     
         82 . The composition of  claim 63 , further comprising lactobionic acid, sodium hydroxide, L-glutathione, potassium chloride, potassium bicarbonate, potassium phosphate, dextrose, sucrose, mannitol, calcium chloride, magnesium chloride, potassium hydroxide, sodium hydroxide, and water. 
     
     
         83 . The composition of  claim 63 , further comprising a cryopreservation medium comprising between about 1% and about 15% DMSO, or between about 0.5% and about 7% DMSO, or between about 1.5% and about 6.5% DMSO, or between about 1.5% and about 3% DMSO, or between about 4% and about 6% DMSO. 
     
     
         84 . The composition of  claim 63 , wherein the cells secrete one or more of the neurotrophic factors: fibroblast growth factors (bFGF and aFGF), ciliary neurotrophic factor (CNTF), pigment epithelium-derived factor (PEDF), brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF). 
     
     
         85 . The composition of  claim 63 , wherein the cells secrete one or more anti-inflammatory cytokines. 
     
     
         86 . A method of treating a retinal condition in a subject in need thereof, the method comprising administering to the subject the composition  claim 63 . 
     
     
         87 . The method of  claim 86 , wherein the retinal condition is a retinal degenerative disease selected from at least one of retinitis pigmentosa, lebers congenital amaurosis, hereditary or acquired macular degeneration, age related macular degeneration (AMD), Best disease, retinal detachment, gyrate atrophy, choroideremia, pattern dystrophy, RPE dystrophies, Stargardt disease, RPE and retinal damage caused by any one of photic, laser, infection, radiation, neovascular or traumatic injury. 
     
     
         88 . The method of  claim 86 , wherein the RPE composition is administered in the subretinal space. 
     
     
         89 . The method of  claim 86 , wherein the composition is delivered with a delivery device comprising a needle with an outer diameter of about 0.63 mm and an inner diameter of about 0.53 mm, a capillary with an outer diameter of about 0.5 mm and an inner diameter of about 0.25 mm, and a tip with an outer diameter of about 0.12 mm and an inner diameter of about 0.07 mm. 
     
     
         90 . The method of  claim 86 , wherein post-delivery percent viability is between about 85% and about 99%, wherein post-delivery percent recovery is between about 65% and about 99%, wherein post-delivery barrier function TEER is between about 100 and about 600Ω, wherein PEDF apical/basal ratio is between about 2 and about 7, and wherein post-delivery VEGF basal/apical ratio is between about 1.5 and about 3. 
     
     
         91 . The method of  claim 86 , wherein the composition is thawed and is injected directly after thawing. 
     
     
         92 . The method of  claim 86 , wherein the composition is administered as a single dose treatment. 
     
     
         93 . The method of  claim 86 , wherein the composition does not cause inflammation after it is administered. 
     
     
         94 . The method of  claim 86 , wherein the cell composition is administered without vitrectomy and without the need to pierce the retina. 
     
     
         95 . The method of  claim 86 , wherein the cell composition is administered by a suprachoroidal injection. 
     
     
         96 . A method of formulating the composition of  claim 63 , comprising:
 a) suspending RPE cells to form a cell suspension in a cell preservation media comprising adenosine, dextran-40, lactobionic acid, HEPES (N-(2-Hydroxyethyl) piperazine-N′-(2-ethanesulfonic acid)), sodium hydroxide, L-glutathione, potassium chloride, potassium bicarbonate, potassium phosphate, dextrose, sucrose, mannitol, calcium chloride, magnesium chloride, potassium hydroxide, sodium hydroxide, dimethyl sulfoxide (DMSO), and water;   b) storing the cell suspension at a cryopreservation temperature; and   c) thawing the cryopreserved suspension, wherein at least about 60% to about 92% of the cells are viable after thawing.