Methods of human retinal progenitor cell isolation and culture
AssigneeThe Regents of the University of California
InventorsJing YANG, Henry KLASSEN
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Abstract
In alternative embodiments, provided herein are methods for isolating primary retinal cells obtained or isolated from a human sample. In alternative embodiments, provided are compositions and methods for treating, ameliorating or preventing a retinal disease or condition; improving a photopic (day light) vision; for improving correcting visual acuity, improving macular function, improving a visual field, or improving scotopic (night) vision by administration of retinal progenitor cells or regenerating a macular and/or a scotopic visual function.
Claims27
A method of isolating primary retinal cells obtained from a human sample by:
- a) processing an obtained sample of human retinal tissues from a human donor of about 12 weeks to about 28 weeks gestational age,
- b) mechanically dissociating the obtained sample,
- c) determining the viability and quantity of the primary retinal cells obtained from the sample, and
- d) confirming the morphology of the obtained primary retinal cells to generate a dissociated suspension of cells and cell clusters.
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A method of culturing isolated primary human retinal cells to produce a population of non-immortal human retinal progenitor cells comprising:
- a) culturing or having cultured a suspension of isolated primary retinal cells in serum-free media in culture flasks or plates coated with a xeno-free fibronectin, an ornithine, a polylysine, or a laminin at standard oxygen levels for between about 4 and 6 passages,
- b) subsequently culturing or having cultured the suspension in serum-free media at low oxygen levels for between about an additional 3 and 6 passages, wherein the cells are passaged at between 40% to 90% confluence and treated with an enzyme at each passage to dissociate the cells and adding fresh culture media, and
- c) subsequently cryopreserving the cells, thereby making a population of non-immortal human retinal progenitor cells.
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A pharmaceutical composition comprising a retinal progenitor cell, or a population or plurality of non-immortal human retinal progenitor cells, isolated by a method of any of the preceding claimsclaim 21, and optionally also comprising a pharmaceutically acceptable excipient.
A kit comprising a retinal progenitor cell, or a population or plurality of non-immortal human retinal progenitor cells, isolated by a method of any of the preceding claimsclaim 21.
A method for:
- treating, ameliorating or preventing a retinal disease or condition,
- improving a photopic (day light) vision,
- improving correcting visual acuity,
- improving macular function,
- improving a visual field, or
- improving scotopic (night) vision,
- the method comprising:
- (a) administering or having administered to an individual in need thereof a retinal progenitor cell, or a population or plurality of non-immortal human retinal progenitor cells, isolated by a method of any of the preceding claimsclaim 21;
- or
- (b) (i) providing or having provided a retinal progenitor cell, or a population or plurality of non-immortal human retinal progenitor cells, isolated by a method of any of the preceding claimsclaim 1;
- and
- (ii) administering or having administered the retinal progenitor cell, or the population or plurality or of non-immortal human retinal progenitor cells to an individual in need thereof.
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Dependencies are inferred from the claim text.