Enriched Pancreatic Stem Cell and Progenitor Cell Populations, and Methods for Identifying, Isolating and Enriching for Such Populations
First Claim
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1. A method of producing a population from pancreatic tissue enriched for human pancreatic stem cells, progenitors, or a combination thereof, comprising:
- a) contacting pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells with a monoclonal antibody that binds CD133 or CD49f; and
b) selecting said pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells that bind to the monoclonal antibody;
wherein the selected cells are enriched for human pancreatic stem cells, progenitors, or a combination thereof.
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Abstract
Enriched pancreatic stem and progenitor cell populations, and methods for identifying, isolating and enriching for pancreatic stem cells using reagents that bind to cell surface markers are provided.
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Citations
52 Claims
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1. A method of producing a population from pancreatic tissue enriched for human pancreatic stem cells, progenitors, or a combination thereof, comprising:
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a) contacting pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells with a monoclonal antibody that binds CD133 or CD49f; and
b) selecting said pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells that bind to the monoclonal antibody;
wherein the selected cells are enriched for human pancreatic stem cells, progenitors, or a combination thereof. - View Dependent Claims (2, 3, 4, 5, 6, 35, 36)
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- 7. A method for producing a population enriched for human pancreatic stem cells, progenitors, or a combination thereof, comprising selecting from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for those cells that are CD133+, or CD49f+, or CD133+CD49f+.
- 14. A method for enriching from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for the populations of pancreatic stem cell or progenitor fractions, comprising selecting from the pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for cells that express CD133 by binding to monoclonal antibody AC133, wherein the selected cells are enriched in the fraction of pancreatic stem cells as compared with the population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells.
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15. A method for enriching a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for the populations of pancreatic stem cell or progenitor cell fractions, comprising selecting from the pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for cells that express CD49f by binding to monoclonal antibody GoH3 or monoclonal antibody 4F10, wherein the selected cells are enriched in the fraction of pancreatic stem cells as compared with the population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells.
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19. A method for isolating a pancreatic stem cell from primary pancreatic tissues, comprising:
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a) selecting from a population of pancreatic cells, pancreatic-derived cells, or gastrointestinal-derived cells for cells that are CD133+, or CD49f+, or CD133+CD49f+;
b) removing the cells that are CD34+, CD45+, or CD34+CD45+, wherein the remaining cells are CD34−
, CD45−
, or CD34−
CD45−
;
c) introducing the cells remaining after step b) to a serum-free culture medium containing one or more growth factors; and
d) proliferating the remaining cells in the culture medium.
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20. A method for producing a population enriched for human pancreatic stem cells, progenitors, or combinations thereof comprising selecting from pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for cells that are CD133+ and bind to an anti-CD133 antibody such as monoclonal antibody AC133, to produce a population enriched for pancreatic stem cells, progenitors, or combinations thereof, wherein the selecting is by attachment to and disattachment from solid phase.
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21. A method for producing a population enriched for human pancreatic stem cells, progenitors, or combinations thereof comprising selecting from pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for cells that are CD49f+ and bind to an anti-CD49f antibody selected from the group consisting of monoclonal antibody GoH3 and monoclonal antibody 4F10, to produce a population enriched for pancreatic stem cells, progenitors, or combinations thereof, wherein the selecting is by attachment to and disattachment from solid phase
- 22. An antibody that specifically binds to the CD49f antigen, wherein said CD49f antigen specifically binds to the monoclonal antibody GoH3 or to the monoclonal antibody 4F10.
- 25. An antibody that specifically binds to the CD133 antigen, wherein said CD133 antigen specifically binds to the monoclonal antibody AC133.
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28. A method for enriching for human pancreatic stem cells, progenitors, or a combination thereof, comprising:
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a) combining a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells with a reagent that specifically binds to the CD133 antigen, the CD49f antigen or both the CD133 and CD49f antigens; and
b) selecting for those cells that bind to the reagent, wherein the selected cells are enriched for pancreatic stem cells, progenitors, or a combination thereof, as compared to the population. - View Dependent Claims (29)
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- 30. A method for producing a population enriched for human pancreatic stem cells, progenitors, or a combination thereof, comprising selecting from pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for those cells that express CD133 and bind to monoclonal antibody AC133, to produce a population enriched for pancreatic stem cells, progenitors, or a combination thereof.
- 31. A method for producing a population enriched for human pancreatic stem cells, progenitors, or a combination thereof, comprising selecting from pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for those cells that express CD49f and bind to monoclonal antibody GoH3 or to monoclonal antibody 4F10, to produce a population enriched for pancreatic stem cells, progenitors, or a combination thereof.
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37. A method for producing a population enriched for human pancreatic stem cells, progenitors, or a combination thereof, wherein the population is obtained from primary pancreas tissues, the method comprising selecting from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for cells that are CD133+, CD49f+, or CD133+CD49f+ the method further comprising the steps of further enriching for pancreatic stem cells, progenitors, or a combination thereof, by further selecting for those cells that are CD34−
- , CD45−
, or CD34−
CD45−
.
- , CD45−
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38. A method for isolating a pancreatic stem cell, comprising:
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a) selecting from a population of primary pancreas tissues, the method comprising selecting from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for at least one selected cell that binds to a monoclonal antibody selected from the group consisting of;
AC133, GoH3, and 4F10;
b) introducing at least one selected cell to a serum free culture medium containing one or more growth factors; and
c) proliferating the at least one selected cell in the culture medium. - View Dependent Claims (39, 40, 41, 42)
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43. A method of producing a population from pancreatic tissue enriched for human pancreatic lineage committed progenitor and mature cells, comprising:
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a) contacting pancreatic cells, pancreatic-derived cells, or gastrointestinal-derived cells with a monoclonal antibody that binds CD9; and
b) selecting said primary pancreas tissues, the method comprising selecting from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells that bind to the monoclonal antibody;
wherein the selected CD9+ cells are enriched for human pancreatic lineage committed progenitor or mature cells capable of differentiating into insulin+ β
cells. - View Dependent Claims (44, 45)
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46. A method for producing a population enriched for human pancreatic lineage committed progenitor cells, comprising selecting from a population of primary pancreas tissues, the method comprising selecting from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells for those cells that are CD49f++CD9+ or CD133+CD9+.
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47. A method for isolating a pancreatic stem cell from primary pancreatic tissues, comprising:
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a) selecting from a population of pancreatic cells, pancreatic-derived cells, or gastrointestinal-derived cells for cells that are CD133+, CD49f+, or CD133+CD49f+;
b) removing the cells that are CD15+ wherein the remaining cells are CD15−
;
c) introducing the cells remaining after step b) to a serum-free culture medium containing one or more growth factors; and
d) proliferating the remaining cells in the culture medium.
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48. A method for producing a population enriched for human pancreatic stem cells, wherein the population is obtained from primary tissues, the method comprising selecting from a population of pancreatic tissue, pancreatic cells, pancreatic-derived cells, or primary gastrointestinal tissue or gastrointestinal-derived cells that are CD133+, CD49f+, or CD133+CD49f+ the method further comprising the steps of further enriching for pancreatic stem cells, by further selecting for those cells that are CD15−
- .
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49. A pancreatic stem cell, wherein the pancreatic stem cell is CD133+CD49f+.
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50. A pancreatic progenitor cell committed to the endocrine β
- -cell lineage, wherein the progenitor cell is CD49f+CD9+.
- View Dependent Claims (51)
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52. A pancreatic progenitor cell committed to the endocrine β
- -cell lineage, wherein the progenitor cell is CD49f+.
Specification