Cell culture methods and media comprising N-acetylcysteine
This application provides improved cell culture media and cell culture methods comprising N-acetylcysteine. These improved cell culture media and cell culture methods increase cell viability, cellular growth rate and/or reduce cell doubling time of cholesterol auxotrophic cells, myeloma cells, and hybridoma cells.
1. A method of increasing cell viability, increasing cell growth rate or reducing cell doubling time comprising:
a. providing a cell culture medium sufficient to support cell growth, wherein the cell culture medium comprises N-acetylcysteine; and
b. culturing a cell in the cell culture medium, wherein the cell is an NS0 cell, wherein the N-acetylcysteine is at a concentration of from 0.25 mM to 3 mM.
2. The method of claim 1 , wherein the cell culture medium is a serum free and animal-protein free medium or is a chemically-defined medium.
3. The method of claim 1 , wherein the average doubling time of the cell is shorter than the average doubling time of a cell in a cell culture with a control medium without N-acetylcysteine, or the average doubling time of the cell is reduced by at least 10%, 15%, 20%, 25% or 50% compared to the control medium without N-acetylcysteine.
4. The method of claim 3 , wherein the average cell doubling time is 60, 42, 32, 30 or 29 hours or less.
5. The method of claim 1 , wherein the cell viability is increased over a cell culture with a control medium excluding N-acetylcysteine, or is increased by at least 5%, 7% or 10% compared to a cell culture medium without N-acetylcysteine, or wherein cell viability is at least 90%, 92% or 93%.
6. The method of claim 1 , wherein the cells express a heterologous protein selected from the group consisting of:
(a) an IL-13 antibody or antigen-binding fragment thereof;
(b) an IL-13 antibody BAK502G9 or an antigen-binding fragment thereof;
(c) an antibody or antigen binding fragment having a heavy chain variable region according to one of SEQ ID NOs: 1, 9, 17, or 25 and a light chain variable region according to one of SEQ ID NOs: 2, 10, 18, or 26; and
(d) an antibody or antigen binding fragment comprising: a heavy chain variable region and a light chain variable region, comprising:
i. a HC CDR1 according to SEQ ID NO: 3, an HC CDR2 according to SEQ ID NO: 4, an HC CDR3 according to SEQ ID NO: 5, an LC CDR1 according to SEQ ID NO: 6, an LC CDR2 according to SEQ ID NO: 7, and an LC CDR3 according SEQ ID NO: 8;
ii. a HC CDR1 according to SEQ ID NO: 11, an HC CDR2 according to SEQ ID NO: 12, an HC CDR3 according to SEQ ID NO: 13, an LC CDR1 according to SEQ ID NO: 14, an LC CDR2 according to SEQ ID NO: 15, and an LC CDR3 according SEQ ID NO: 16;
iii. a HC CDR1 according to SEQ ID NO: 19, an HC CDR2 according to SEQ ID NO: 20, an HC CDR3 according to SEQ ID NO: 21, an LC CDR1 according to SEQ ID NO: 22, an LC CDR2 according to SEQ ID NO: 23, and an LC CDR3 according SEQ ID NO: 24; or
iv. a HC CDR1 according to SEQ ID NO: 27, an HC CDR2 according to SEQ ID NO: 28, an HC CDR3 according to SEQ ID NO: 29, an LC CDR1 according to SEQ ID NO: 30, an LC CDR2 according to SEQ ID NO: 31, and an LC CDR 3 according SEQ ID NO: 32.