Method for identification of cell growth or differentiation factors
First Claim
Patent Images
1. A cell population selected from the group consisting of (a) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of Fcγ
- RII, Fcγ
RIII, Thy-1, CD44, VLA-4α
, LFA-1β and
combinations thereof;
(b) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of Fcγ
RII, Fcγ
RIII, CD44, VLA-4α
, LFA-1β and
combinations thereof;
(c) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of HSA, CD44, VLA-4α
, LFA-1β
, ICAM-1 and combinations thereof;
(d) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of CD45, Aa4.1, Sca-1, HSA, Fcγ
Fcγ
, Thy-1 Mac-1, Gr-1, CD44, VLA-4α
, LFA-1β and
combinations, thereof; and
(e) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of CD45, Aa4.1, HSA, Fcγ
RII, Fcγ
RIII, Thy-1, Mac-1, Gr-1, CD44, VLA-4α
, LFA-1β
, ICAM-1 and combinations thereof.
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Abstract
The present invention relates to novel immortalized precurso cell populations derived from embryonic stem cell populations and methods to produce such cell populations. Also disclosed is an assay to identify regulatory compounds capable of controlling cell growth for therapeutic and experimental use.
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Citations
83 Claims
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1. A cell population selected from the group consisting of (a) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of Fcγ
- RII, Fcγ
RIII, Thy-1, CD44, VLA-4α
, LFA-1β and
combinations thereof;
(b) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of Fcγ
RII, Fcγ
RIII, CD44, VLA-4α
, LFA-1β and
combinations thereof;
(c) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of HSA, CD44, VLA-4α
, LFA-1β
, ICAM-1 and combinations thereof;
(d) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of CD45, Aa4.1, Sca-1, HSA, Fcγ
Fcγ
, Thy-1 Mac-1, Gr-1, CD44, VLA-4α
, LFA-1β and
combinations, thereof; and
(e) a HOX11 precursor cell population comprising a cell surface molecule selected from the group consisting of CD45, Aa4.1, HSA, Fcγ
RII, Fcγ
RIII, Thy-1, Mac-1, Gr-1, CD44, VLA-4α
, LFA-1β
, ICAM-1 and combinations thereof. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
- RII, Fcγ
- 18. A HOX11 precursor cell population that is responsive to a growth factor selected from the group consisting of interleukin-3, interleukin-4, interleukin-6, interleukin-11, erythropoietin, C-kit ligand, leukocyte inhibitory factor and mixtures thereof.
- 22. A HOX11 precursor cell population that is responsive to a growth factor selected from the group consisting of interleukin-3, erythropoietin, C-kit ligand, leukocyte inhibitory factor and mixtures thereof.
- 26. A HOX11 precursor cell population that is responsive to a growth factor selected from the group consisting of interleukin-3, erythropoietin, GM-CSF and mixtures thereof.
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29. A precursor cell population having the identifying characteristics of a cell population selected from the group consisting of EBHX-1, EBHX-2, EBHX-3, EBHX-4, EBHX-5, EBHX-6, EBHX-7, EBHX-8, EBHX-9, EBHX-10, EBHX-11, EBHX-12, EBHX-13, EBHX-14 and EBHX-15.
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30. A method to produce an immortalized precursor cell population, comprising:
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a) transforming an embryonic stem cell population with an immortalizing gene to create a transformed stem cell population;
b) culturing said transformed stem cell population under effective conditions to produce a transformed embryoid body cell population; and
c) incubating said transformed embryoid body cell population under conditions suitable to obtain an immortalized precursor cell population. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37)
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38. A method to identify a regulatory factor that influences the growth or differentiation of a cell, comprising:
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a) contacting a HOX11 precursor cell population with a regulatory factor selected from the group consisting of a putative regulatory factor, a known regulatory factor and mixtures thereof; and
b) assessing the responsiveness of said precursor cell population to said regulatory factor. - View Dependent Claims (39, 40, 41, 42, 43)
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- 44. A method to identify a compound expressed during the development of a population of embryonic stem cells, comprising characterizing at least a portion of the cellular composition of at least one cell contained in a HOX11 precursor cell population, to identify a compound expressed during the development of a population of embryonic stem cells.
- 46. A method to produce an antibody, comprising administering to an animal an effective amount of a protein derived from a HOX11 precursor cell population and recovering an antibody capable of selectively binding to said protein.
- 48. A method to repopulate a hematopoietic cell population in an animal, comprising administering to an animal a suitable number of cells of a HOX11 precursor cell population.
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50. A method to identify a neutralizing reagent, comprising:
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a) contacting a HOX11 precursor cell population with a known regulatory factor to produce a controlled cell population;
b) combining said controlled cell population with a neutralizing reagent selected from the group consisting of a known neutralizing compound of said regulatory factor and a putative neutralizing compound of said regulatory factor; and
c) assessing the responsiveness of said precursor cell population to said neutralizing compound.
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51. An endothelial cell population produced by the method comprising,
a) transforming an EB cell population with a nucleic acid molecule encoding a Polyoma Middle T antigen to form Polyoma Middle T EB cells; - and
b) culturing said Polyoma Middle T EB cells under conditions suitable to obtain an endothelial cell population. - View Dependent Claims (52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62)
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63. An endothelial cell population having the identifying characteristics of D4T.
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64. A conditioned medium. comprising a cell culture supernatant recovered from a culture of an endothelial cell population produced by the method comprising,
a) transforming an EB cell population with a nucleic acid molecule encoding a Polyoma Middle T antigen to form Polyoma Middle T EB cells; - and
b) culturing said Polyoma Middle T EB cells under conditions suitable to obtain an endothelial cell population. - View Dependent Claims (65, 66, 67, 68, 69)
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70. A method to identify a hematopoietic cell growth factor, comprising isolating a compound from a conditioned medium comprising a cell culture supernatant recovered from a culture of an endothelial cell population produced by the method comprising,
a) transforming an EB cell population with a nucleic acid molecule encoding a Polyoma Middle T antigen to form Polyoma Middle T EB cells; - and
b) culturing said Polyoma Middle T EB cells under conditions suitable to obtain an endothelial cell population. - View Dependent Claims (71, 72, 73)
- and
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74. A method to enhance a population of precursor cells, comprising contacting a precursor cell population with a cell culture supernatant recovered from a culture of an endothelial cell population produced by the method comprising,
a) transforming an EB cell population with a nucleic acid molecule encoding a Polyoma Middle T antigen to form Polyoma Middle T EB cells; - and
b) culturing said Polyoma Middle T EB cells under conditions suitable to obtain an endothelial cell population. - View Dependent Claims (75)
- and
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76. An enhanced precursor cell population, comprising a precursor cell population contacted with a cell culture supernatant recovered from a culture of an endothelial cell population produced by the method comprising,
a) transforming an EB cell population with a nucleic acid molecule encoding a Polyoma Middle T antigen to form Polyoma Middle T EB cells; - and
b) culturing said Polyoma Middle T EB cells under conditions suitable to obtain an endothelial cell population, wherein said precursor cell population, when contacted with said cell supernatant results in the formation of an enhanced precursor population. - View Dependent Claims (77, 78, 79, 80)
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81. A method to identify a hematopoietic cell growth factor, comprising isolating a compound from an endothelial cell population produced by the method comprising,
a) transforming an EB cell population with a nucleic acid molecule encoding a Polyoma Middle T antigen to form Polyoma Middle T EB cells; - and
b) culturing said Polyoma Middle T EB cells under conditions suitable to obtain an endothelial cell population. - View Dependent Claims (82, 83)
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Specification