Tissue system with undifferentiated stem cells derived from corneal limbus
First Claim
1. A tissue system comprising limbal stem cells, wherein at least about 30-90% of the limbal stem cells in the tissue system are undifferentiated stem cells.
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Abstract
The present disclosure describes a tissue system with self-regenerating limbal stem cells, wherein the limbal stem cells are primarily undifferentiated stem cells (USCs). The tissue system is derived from isolated corneal limbal tissue, and is suitable for restoring ocular surface impairments, particularly those that result from limbal stem cell deficiencies. The tissue system is generated by selectively augmenting the tissue system for USCs, for example by selecting and sorting cells that express stem cell-specific surface markers, such as stage specific embryonic antigen marker 4 (SSEA-4). After isolation, the USCs are cultured on a tissue base in the presence of enriched medium to generate the tissue system, which is suitable for transplantation, implantation, or grafting.
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48 Claims
- 1. A tissue system comprising limbal stem cells, wherein at least about 30-90% of the limbal stem cells in the tissue system are undifferentiated stem cells.
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10. A method of generating a tissue system, wherein the tissue system comprises limbal stem cells, comprising the steps of:
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(a) isolating corneal limbal tissue from a donor;
(b) culturing the corneal limbal tissue to expand corneal limbal cells in culture;
(c) isolating a population of limbal stem cells from the cultured corneal limbal cells by sorting the corneal limbal cells to select for one or more stem cell-specific surface markers, wherein the stem cell-specific surface marker is expressed by undifferentiated stem cells;
(d) culturing the isolated population of limbal stem cells to generate the tissue system. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48)
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Specification