CELL CULTURE MEDIUM

1 .- 146 . (canceled)

147 . A method comprising culturing a population of genetically modified immune effector cells or hematopoietic stem or progenitor cells (HSPCs) in a culture medium comprising L-ornithine, wherein the culture medium has an osmolarity of about 275 mOsm/kg to about 320 mOsm/kg, and

wherein cell proliferation is increased compared to cells grown in a culture medium without L-ornithine and/or an osmolality of about 275 mOsm/kg to about 320 mOsm/kg.

148 . A method of expanding a population of genetically modified immune effector cells or hematopoietic stem or progenitor cells (HSPCs), comprising culturing the cells in a culture medium comprising L-ornithine, wherein the culture medium has an osmolarity of about 275 mOsm/kg to about 320 mOsm/kg, and

wherein cell proliferation is increased compared to cells grown in a culture medium without L-ornithine and/or an osmolality of about 275 mOsm/kg to about 320 mOsm/kg.

149 . A method of manufacturing a population of genetically modified immune effector cells or hematopoietic stem or progenitor cells (HSPCs), comprising culturing the cells in a culture medium comprising L-ornithine, wherein the culture medium has an osmolarity of about 275 mOsm/kg to about 320 mOsm/kg, and

wherein cell proliferation is increased compared to cells grown in a culture medium without L-ornithine and/or an osmolality of about 275 mOsm/kg to about 320 mOsm/kg.

150 . The method of claim 149 , wherein cell viability is increased compared to cells grown in a culture medium without L-ornithine and/or an osmolality of about 275 mOsm/kg to about 320 mOsm/kg.

151 . The method of claim 149 , wherein CD62L+ expression is increased in the genetically modified immune effector cells compared to genetically modified immune effector cells grown in a culture medium without L-ornithine and/or an osmolality of about 275 mOsm/kg to about 320 mOsm/kg.

152 . The method of claim 149 , wherein the genetically modified cells are modified to express a therapeutic protein, and wherein the therapeutic protein expression is increased compared to genetically modified cells grown in a culture medium without L-ornithine and/or an osmolality of about 275 mOsm/kg to about 320 mOsm/kg.

153 . The method of claim 149 , wherein the cells are cultured at about 38° C. to about 39.5° C.

154 . The method of claim 149 , wherein the culture medium has an osmolarity of about 310 mOsm/kg.

155 . The method of claim 149 , wherein the culture medium comprises about 0.75 to about 3.0 mM L-ornithine.

156 . The method of claim 149 , wherein the culture medium comprises a recombinant growth factor that increases ornithine decarboxylase (ODC) expression and/or activity.

157 . The method of claim 156 , wherein the recombinant growth factor is IL-2.

158 . The method of claim 149 , wherein:

a) the culture medium comprises a final ratio of NaCl to KCl of about 20:1 to about 30:1;

b) the culture medium comprises about 0.5 mM to about 3 mM CaCl 2 ; and/or

c) the culture medium comprises about 0.40 g/kg to about 0.80 g/kg NaHCO 3 .

159 . The method of claim 149 , wherein the culture medium comprises about 0.5 g/kg to about 1.5 g/kg poloxamer 188.

160 . The method of claim 149 , wherein the culture medium comprises about 5 mM to about 25 mM HEPES.

161 . The method of claim 149 , wherein the culture medium comprises about 40 g/L to about 60 g/L heat inactivated (HI) AB serum or gamma irradiated (GI) AB serum.

162 . The method of claim 149 , wherein the culture medium comprises a base medium for culturing immune effector cells and/or hematopoietic stem or progenitor cells.

163 . The method of claim 162 , wherein the base medium is X-VIVO 15 or IMDM, or variant thereof.

164 . The method of claim 149 , wherein the culture medium further comprises:

a) recombinant GM-CSF, G-CSF, IFN-γ, TGFβ, and/or TNFα;

b) human serum albumin (HSA);

c) cholesterol; and/or

d) vitamin E.

165 . The method of claim 149 , wherein the population of immune effector cells comprises cytotoxic T lymphocytes (CTLs), helper T cells, natural killer (NK) cells, natural killer T (NKT) cells, regulatory T cells, or dendritic cells.

166 . The method of claim 149 , wherein the population of hematopoietic stem or progenitor cells (HSPCs) are CD44+, CD34+, CD90+ and/or CD133+ cells.

167 . A medium for culturing a population of genetically modified immune effector cells and/or hematopoietic stem or progenitor cells (HSPCs) comprising:

a) a base medium for culturing genetically modified immune effector cells or hematopoietic stem or progenitor cells (HSPCs),

b) a buffer that maintains the pH of the culture medium in the range of about 6.5 to about 7.5,

c) a serum or suitable serum replacement,

d) one or more mono- and di-valent salts, wherein the mono- and di-valent salts comprise CaCl 2 , NaCl, and/or KCl, and further wherein the final molar ratio of NaCl to KCl in the culture medium is about 20:1 to about 30:1,

e) a recombinant growth factor, or chemically defined replacement, that increases ornithine decarboxylase (ODC) expression and/or activity; and

f) about 0.75 to about 3.0 mM L-ornithine;

wherein the cell culture medium has an osmolarity of about 275 mOsm/kg to about 320 mOsm/kg.

168 . The medium of claim 167 , wherein the culture medium comprises:

a) a base medium comprising X-VIVO 15,

b) about 10 mM HEPES buffer,

c) about 50 g/L HI Human AB Serum or GI AB serum,

d) about 0.60 g/kg NaHCO 3 ,

e) about 1.97 mM CaCl 2 , about 4.585 g/L NaCl, and about 0.33 g/L KCl, wherein the final molar ratio of NaCl to KCl in the culture medium is about 28:1,

f) about 1 g/kg poloxamer 188,

g) about 0.43 g/L L-alanine-L-glutamine dipeptide,

h) about 250 IU/mL recombinant human IL-2 growth factor; and

i) about 0.25 g/kg L-ornithine HCl;

wherein the cell culture medium has an osmolarity of about 310 mOsm/kg.