Methods for Generating Inner Ear Cells in Vitro
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsStefan Heller, Mohammad Ronaghi, Kazuo Oshima
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Abstract
Methods, compositions and kits are provided for generating inner ear cells in vitro. These methods find use a number of applications, such as in preparing inner ear cells for in vitro screening for agents that are toxic to inner ear cells, for in vitro screening for agents that prevent against, mitigate, or reverse the toxic effects of such agents, and for in vitro screening for agents that promote otoregeneration.
Claims23
A method of generating inner ear cells from pluripotent stem cells in vitro, the method comprising:
- a. culturing pluripotent stem cells in the presence of at least one factor that suppresses the formation of endoderm and mesoderm and at least one ectoderm rostralizing factor, wherein a population comprising preplacodal ectodermal cells is produced;
- b. culturing the population comprising preplacodal ectodermal cells under adherent conditions in the presence of at least one FGF, wherein a population comprising otic progenitor cells is produced;
- and
- c. culturing the population comprising otic progenitor cells under adherent conditions, wherein a population comprising inner ear cells is produced.
A method of generating human inner ear cells in vitro, the method comprising:
- culturing a population of cells that are enriched for otic progenitor cells under adherent conditions in the absence of feeder cells to form inner ear cells.
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A method of screening a candidate agent for an effect on inner ear cells, the method comprising:
- contacting inner ear cells generated from pluripotent stem cells in vitro with a candidate agent;
- and
- comparing the viability, function and/or regenerative response of the inner ear cells contacted with the candidate agent to the viability, function and/or regenerative response of inner ear cells that were not contacted with the candidate agent;
- wherein a change in viability, function and/or regenerative response of inner ear cells contacted with the candidate agent indicates that the candidate agent has an effect on inner ear cells.
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Dependencies are inferred from the claim text.