MEDIA AND METHODS FOR DIFFERENTIATING NATURAL KILLER CELLS
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.
1 . An NK cell differentiation medium, comprising a pyrimidoindole compound.
2 . The NK cell differentiation medium according to claim 1 , wherein the pyrimidoindole compound is UM171 or UM729.
3 . The NK cell differentiation medium according to claim 1 , further comprising a basal medium.
4 . The NK cell differentiation medium according to claim 3 , further comprising one or more of SCF, FLT3L, IL-2, IL-3, IL-15, or IL-7.
5 . The NK cell differentiation medium according to claim 1 , wherein the NK cell differentiation medium is not conditioned by contact with stromal cells or a stroma cell replacement.
6 . The NK cell differentiation medium according to claim 1 , wherein the NK cell differentiation medium does not include an aryl hydrocarbon receptor antagonist.
7 . The NK cell differentiation medium according to claim 1 , wherein the NK cell differentiation medium is serum-free.
8 . The NK cell differentiation medium according to claim 1 , wherein the NK cell differentiation medium differentiates NK cell progenitors.
9 . The NK cell differentiation medium according to claim 8 , wherein the NK cell progenitors are isolated or derived from a primary sample, or are derived from pluripotent stem cells.
10 - 11 . (canceled)
12 . A method for differentiating NK cells, comprising:
providing a population of NK cell progenitors; and
contacting the population of NK cell progenitors in culture with the medium of claim 1 at a concentration and for a time sufficient to yield NK cells,
wherein the NK cells express CD56.
13 . The method according to claim 12 , wherein the population of NK cell progenitors express CD7 or CD5, or both.
14 - 15 . (canceled)
16 . The method according to claim 12 , wherein the NK cells increase in frequency during the contacting step or increase in number during the contacting step, or both.
17 . (canceled)
18 . The method according to claim 12 , wherein the population of NK cell progenitors increase or decrease in frequency or number during the contacting step.
19 . The method according to claim 12 , wherein the providing and contacting steps are not in the presence of stromal cells or a stroma cell replacement.
20 . The method according to claim 12 , wherein the providing and contacting steps are not in the presence of an aryl hydrocarbon receptor antagonist.
21 . The method according to claim 12 , wherein the concentration of the pyrimidoindole compound is between 10 nM and 3 μM.
22 . The method according to claim 12 , wherein the time is at least 1 week or about 2 weeks.
23 - 24 . (canceled)
25 . The method according to claim 12 , wherein the NK cells are cytotoxic.
26 . The method according to claim 12 , wherein the population of NK cell progenitors is derived or isolated from a primary sample.
27 - 28 . (canceled)
29 . The method according to claim 12 , wherein the population of NK cells is differentiated from a pluripotent stem cell.