OPTIMIZATION OF NK-92 CELL GROWTH USING POLOXAMER
Provided herein are methods of culturing NK-92® cells using the growth media containing a non-ionic surfactant such that the cell culture have reduced clumping as compared to control NK-92® cells that have been cultures in a control medium lacking the non-ionic surfactant. The growth medium comprises 0.025 to 0.9% of a non-ionic surfactant, e.g., Poloxamer 188.
1 . A method of culturing NK-92® cells comprising culturing the NK-92® cells in a culture medium comprising 0.025% to 0.9% of a non-ionic surfactant, wherein the NK-92® cell culture has reduced clumping as compared to control NK-92® cells that have been cultured in a control medium lacking the non-ionic surfactant.
2 . The method of claim 1 , wherein the NK-92® cells maintained the substantially the same cytotoxicity as the control NK-92® cells.
3 . The method of claim 1 , wherein the cell culture is substantially free from clumping.
4 . The method of claim 1 , wherein the cells are cultured in at least 2 liters of culture medium.
5 . The method of claim 1 , wherein the non-ionic surfactant is Poloxamer 188.
6 . The method of claim 5 , wherein the culture medium comprises from 0.025% to 0.06% of Poloxamer 188.
7 . The method of claim 5 , wherein the culture medium comprises 0.05% of the Poloxamer 188.
8 . The method of claim 1 , wherein NK-92® cell culture has reduced cell aggregates as compared to a control cell culture.
9 . The method of claim 8 , wherein reduction of the percentage of cell aggregates is at least 40%.
10 . The method of claim 8 , wherein the NK-92® cell culture has less than 6% cell aggregates after 3 days of culturing.
11 . The method of claim 1 , wherein the NK-92® cells have a viability of at least 80%.
12 . The method of claim 1 , wherein the NK-92® cells comprise a cytokine, Fc Receptor, chimeric antigen receptor or a combination thereof.
13 . A method of reducing fluocculants in a culture medium, the method comprising adding to the culture medium 0.025% to 0.9% of a non-ionic surfactant, wherein the culture medium has reduced fluocculants as compared to control culture lacking the non-ionic surfactant.
14 . A cell culture comprising NK-92® cells and a culture medium comprising 0.025% to 0.9% of the non-ionic surfactant, wherein the cell culture has reduced clumping as compared to control culture comprising NK-92 cells and a medium lacking the non-ionic surfactant.
15 . The cell culture of claim 14 , wherein the NK-92® cells have been cultured for at least 3 days.
16 . The cell culture of claim 14 , wherein the cell culture is substantially free from clumping.
17 . The cell culture of claim 14 , wherein the NK-92® cells maintained the substantially the same cytotoxicity as the NK-92 cells in the control culture.
18 . The cell culture of claim 14 , wherein the non-ionic surfactant is Poloxamer 188.
19 . The cell culture of claim 14 , wherein the cell culture comprises 0.025% to 0.06% of the Poloxamer 188.
20 . The cell culture of claim 14 , wherein the cell culture comprises 0.05% of the Poloxamer 188.
21 . The cell culture of claim 14 , wherein the NK-92® cells comprise a cytokine, Fc Receptor, chimeric antigen receptor, or a combination thereof.
22 . The cell culture of claim 14 , wherein the cell culture has a volume of at least 2 liters.
23 . The cell culture of claim 22 , wherein the cell culture has a volume of at least 10 liters.