Method for production of neutrophils and uses therefor
First Claim
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1. A method for producing neutrophils in vitro, comprising:
- a) providing an expanded population of neutrophil progenitor cells;
b) culturing the expanded population of neutrophil progenitor cells with semi-confluent stromal cells in a medium suitable for culture of animal cells, the medium comprising at least one interleukin-6 (IL-6) family cytokine, to produce a secondary differentiation culture; and
c) culturing the cells from the secondary differentiation culture of step (b) with semi-confluent stromal cells in a medium suitable for culture of animal cells, the medium comprising;
granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GM-CSF) and at least one IL-6 family cytokine, to produce functionally mature neutrophils.
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Abstract
An in vitro method for the differentiation of functionally mature neutrophils from stem cells is disclosed. Also disclosed are methods of producing genetically modified neutrophils in vitro and methods of using the neutrophils produced according to the invention.
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Citations
53 Claims
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1. A method for producing neutrophils in vitro, comprising:
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a) providing an expanded population of neutrophil progenitor cells;
b) culturing the expanded population of neutrophil progenitor cells with semi-confluent stromal cells in a medium suitable for culture of animal cells, the medium comprising at least one interleukin-6 (IL-6) family cytokine, to produce a secondary differentiation culture; and
c) culturing the cells from the secondary differentiation culture of step (b) with semi-confluent stromal cells in a medium suitable for culture of animal cells, the medium comprising;
granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GM-CSF) and at least one IL-6 family cytokine, to produce functionally mature neutrophils. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 46, 47, 48, 49, 50, 51, 53)
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29. The method of claim 29, wherein the hydrocortisone is 21-hemisuccinate sodium salt.
- 45. An isolated, genetically modified neutrophil, wherein the neutrophil is produced in vitro.
Specification