NATURAL KILLER CELLS AND USES THEREOF
Provided herein are methods of producing natural killer (NK) cells and NK progenitor cell populations using a two-step expansion and differentiation method. Also provided herein are methods of producing populations of NK cells and NK progenitor cell populations using a three-step expansion and differentiation method. Also provided herein are methods of suppressing tumor cell proliferation using the NK cells, the NK cell populations, and the NK progenitor cell populations produced by the methods described herein, as well as methods of treating individuals having cancer or a viral infection, comprising administering the NK cells, the NK cell populations, and the NK progenitor cell populations produced by the methods described herein to an individual having the cancer or viral infection.
1 . An isolated population of natural killer (NK) progenitor cells, wherein said NK progenitor cells are produced according to the following method: (i) culturing hematopoietic stem cells or progenitor cells in a first medium comprising Flt3L, TPO, SCF, IL-7, G-CSF, IL-6 and GM-CSF, (ii) subsequently culturing said cells in a second medium comprising Flt3L, SCF, IL-15, and IL-7, IL-17 and IL-15, G-CSF, IL-6 and GM-CSF, and (iii) subsequently culturing said cells in a third medium comprising SCF, IL-15, IL-7, IL-2, G-CSF, IL-6 and GM-CSF.
2 . The isolated population of NK progenitor cells of claim 1 , wherein the duration of culturing step (i) is 7-9 days, wherein the duration of culturing step (ii) is 5-7 days, and wherein the duration of culturing step (iii) is 5-9 days.
3 . The isolated population of NK progenitor cells of claim 1 , wherein the duration of culturing step (i) is 7-9 days, wherein the duration of culturing step (ii) is 5-7 days, and wherein the duration of culturing step (iii) is 21-35 days.
4 . The isolated population of NK progenitor cells of claim 1 , 2 , or 3 wherein the hematopoietic stem or progenitor cells used in the method are CD34 + .
5 . The isolated population of NK progenitor cells of claim 1 , 2 , or 3 wherein the hematopoietic stem or progenitor cells comprise hematopoietic stem or progenitor cells from human placental perfusate and hematopoietic stem or progenitor cells from umbilical cord, wherein said placental perfusate and said umbilical cord blood are from the same placenta.
6 . The isolated population of NK progenitor cells of any one of claims 1 - 5 , wherein CD34− cells comprise more than 80% of the total population at the end of step (i).
7 . The isolated population of NK progenitor cells of any one of claims 1 - 6 , wherein said population comprises no more than 40% CD3−CD56+ cells
8 . The isolated population of NK progenitor cells of any one of claims 1 - 7 , wherein said population comprises cells which are CD52+CD117+.
9 . An isolated natural killer (NK) progenitor cell population, wherein said population comprises no more than 40% CD3−CD56+ cells.
10 . An isolated natural killer (NK) progenitor cell population, wherein said population comprises cells which are CD52+CD117+.
11 . A pharmaceutical composition comprising the isolated population of NK progenitor cells of any one of claims 1 - 10 .
12 . A method of suppressing the proliferation of tumor cells comprising bringing the tumor cells into proximity with the isolated population of NK progenitor cells of any one of claims 1 - 10 or the composition of claim 11 .
13 . The method of claim 12 , wherein said tumor cells are primary ductal carcinoma cells, glioblastoma cells, leukemia cells, acute T cell leukemia cells, chronic myeloid lymphoma (CML) cells, acute myelogenous leukemia cells, chronic myelogenous leukemia (CML) cells, lung carcinoma cells, colon adenocarcinoma cells, histiocytic lymphoma cells, multiple myeloma cells, colorectal carcinoma cells, colorectal adenocarcinoma cells, prostate cancer cells, or retinoblastoma cells.
14 . A method of treating hematologic cancer in a subject in need thereof, comprising administering to said subject the isolated population of NK progenitor cells of any one of claims 1 - 10 or the composition of claim 11 .
15 . The method of claim 14 , wherein said hematologic cancer is acute myeloid leukemia.