Myeloma cell culture in transferrin-free low iron medium
The present invention relates to a method for culturing mammalian cells in a culture medium which is transferrin free and which contains no lipophilic or synthetic nitrogen-containing chelators. Also provided is the use of the medium and a process for providing a mammalian product by culturing cells capable of producing the product in the medium.
1. A method for the in vitro culture of a myeloma cell line, said method comprising:
(a) inoculating a culture medium with the myeloma cell line, said medium being capable of supporting the growth of said myeloma cell line and comprising iron at concentrations in the medium of from 0.064 mg/L to 1.6 mg/L, wherein said medium does not contain any of a transferrin, a lipophilic chelator, a synthetic nitrogen-containing chelator and a lipophilic synthetic nitrogen-containing chelator; and
(b) growing of the inoculated culture medium under appropriate conditions and using agitated suspension culture, and
wherein the source of iron is a soluble iron compound selected from the group consisting of ferric ammonium citrate, ferric ammonium oxalate, ferric ammonium fumarate, ferric ammonium malate and ferric ammonium succinate.
2. The method of claim 1 wherein the concentration of iron in the medium is from about 0.16 mg/L to about 0.32 mg/L.
3. The method of claim 1 wherein the source of iron is ferric ammonium citrate.
4. The method of claim 1 wherein the medium is serum free, protein free, free of components of animal derivation or is chemically defined.
5. The method of claim 1 wherein the myeloma cell line is selected from the group consisting of an NSO series cell line, a P3 series cell line, MOPC series cell line, the MPC-11 cell line, the J558L cell line, the K6H6/B5 cell line, the 45.6.TG1.7 cell line, the YO cell line, the Y3 HTK cell line, the RPMI 8226 cell line and the U266B1 cell line.
6. The method of claim 1 wherein the myeloma cell line is the NSO cell line.
7. A method for the in vitro culture of a myeloma cell line, the method comprising:
(a) inoculating a culture medium with the myeloma cell line, said medium being capable of supporting the growth of said myeloma cell line and comprising ferric ammonium citrate at a concentration in the medium of from 0.4 mg/L to 10 mg/L, wherein said medium does not contain any of a transferrin, a lipophilic chelator, a synthetic nitrogen-containing chelator and a lipophilic synthetic nitrogen-containing chelator; and
(b) growing of the inoculated culture medium under appropriate conditions and using agitated suspension culture.
8. The method of claim 7 wherein the ferric ammonium citrate is present in the medium at a concentration of from about 1 mg/L to about 2 mg/L.
9. A process for obtaining a mammalian cell product comprising culturing a myeloma cell capable of producing said product under agitated suspension culture and in a culture medium capable of supporting the growth of said myeloma cell line, said medium comprising iron at concentrations in the medium of from 0.064 mg/L to 1.6 mg/L, or ferric ammonium citrate at a concentration in the medium of from about 0.4 mg/L to about 10 mg/L, wherein said medium does not contain any of a transferrin, a lipophilic chelator, a synthetic nitrogen-containing chelator and a lipophilic synthetic; nitrogen-containing chelator; and recovering said mammalian cell product, and
wherein the source of iron is a soluble iron compound selected from the group consisting of ferric ammonium citrate, ferric ammonium oxalate, ferric ammonium fumarate, ferric ammonium malate and ferric ammonium succinate.
10. The process of claim 9 wherein the concentration of iron in the medium is from about 0.16 mg/L to about 0.32 mg/L.
11. The process of claim 9 wherein the source of iron is ferric ammonium citrate.
12. The process of claim 9 wherein the ferric ammonium citrate is present in the medium at a concentration of from about 1 mg/L to about 2 mg/L.
13. The process of claim 9 wherein the medium is serum free, protein free, free of components of animal derivation or is chemically defined.
14. The process of claim 9 wherein the myeloma cell line is selected from the group consisting of an NSO series cell line, a P3 series cell line, a MOPC series cell line, the MPC-11 cell line, the J558L cell line, the K6H6/B5 cell line, the 45.6.TG1.7 cell line, the YO cell line, the Y3 HTK cell line, the RPMI 8226 cell line and the U266B1 cell line.
15. The process of claim 9 wherein the myeloma cell line is the NSO cell line.
16. The process of claim 9 wherein the cell product is at least one product selected from the group consisting of a polypeptide, a protein, a hormone, a lymphokine, an interleukin, an industrially useful enzyme and a therapeutically useful enzyme.
17. The process of claim 16 wherein the cell product is an antibody or fragment thereof.