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US 12,723,233 B2Granted
Media and methods for differentiating natural killer cells
AssigneeStemcell Technologies Canada Inc.
InventorsNooshin Tabatabaei-Zavareh, Albertus Wernerus Wognum
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Abstract
Media and methods for differentiating NK progenitor cells into NK cells are disclosed. The NK differentiation media comprises a pyrimidoindole compound such as UM171 or UM729.
Claims13
A method for differentiating NK cells, comprising:
- providing a population of NK cell progenitors, wherein the population of NK cell progenitors arise in contact with a NK cell progenitor differentiation medium and a stroma cell replacement comprising components selected from fibronectin, whether full-length or a fragment thereof, VCAM-1, and an immobilized Notch ligand;
- and
- contacting the population of NK cell progenitors in culture with an NK cell differentiation medium comprising a basal medium, IL-15 and a pyrimidoindole compound at a concentration between 10 nM and 3 μM and for a time sufficient to yield NK cells,
- wherein the NK cells co-express i) CD56, NKG2D and NKp44, or ii) CD56, NKG2D and NKp30,
- wherein an increased frequency and/or yield of NK cells co-expressing i) CD56, NKG2D and NKp44, or ii) CD56, NKG2D and NKp30 is obtained when contacting the population of NK cell progenitors with the NK cell differentiation medium as compared to when the NK cells are differentiated from the population of NK cell progenitors in a NK cell differentiation medium devoid of an added pyrimidoindole compound, and
- wherein the NK cells produce intracellular interferon γ upon stimulation.
3 more elements
A method for differentiating NK cells, comprising:
- providing a population of NK cell progenitors, wherein the population of NK cell progenitors arise in contact with a NK cell progenitor differentiation medium and a stroma cell replacement comprising components selected from fibronectin, whether full-length or a fragment thereof, VCAM-1, and an immobilized Notch ligand;
- and
- contacting the population of NK cell progenitors in culture with an NK cell differentiation medium comprising a basal medium, IL-15 and a pyrimidoindole compound at a concentration between 10 nM and 3 μM and for a time sufficient to yield NK cells,
- wherein the NK cells co-express i) CD56, NKG2D and NKp44, or ii) CD56, NKG2D and NKp30,
- wherein an increased frequency and/or yield of NK cells co-expressing i) CD56, NKG2D and NKp44, or ii) CD56, NKG2D and NKp30 is obtained when contacting the population of NK cell progenitors with the NK cell differentiation medium as compared to when the NK cells are differentiated from the population of NK cell progenitors in an NK cell differentiation medium devoid of an added pyrimidoindole compound,
- wherein less than 5% of the NK cells express CD3, and
- wherein the NK cells produce intracellular interferon γ upon stimulation.
4 more elements
Dependencies are inferred from the claim text.