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1. WO2020142304 - SUPPLEMENTED SERUM-CONTAINING CULTURE MEDIUM FOR ENHANCED ARPE-19 GROWTH AND HUMAN CYTOMEGALOVIRUS VACCINE PRODUCTION

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[ EN ]

WHAT IS CLAIMED IS:

1. A cell culture medium comprising a basal cell culture medium that comprises serum and is further supplemented with between about 0.1 mM and about 10 mM of dexamethasone, between about 0.1 ng/mL and about 100 ng/mL of epidermal growth factor (EGF), IX MEM essential amino acids, 1X-4X glutamine synthetase expression medium (GSEM) supplement and trace elements.

2. The medium of claim 1, wherein the basal cell culture medium is

DMEM/F-12.

3. The medium of claim 1 or 2, wherein said serum is fetal bovine serum

(FBS).

4. The medium of claim 3, wherein the FBS is present at a concentration between about 2% and about 10%, inclusive.

5. The medium of any one of claims 1-4, wherein the dexamethasone is present at a concentration between about 1 and about 10 mM.

6. The medium of any one of claims 1-5, wherein the EGF is present at a concentration between about 1 and about 40 ng/mL.

7. The medium of any one of claims 1 to 6, wherein the IX MEM essential amino acids consist of arginine, cysteine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, and valine.

8. The medium of any one of claims 1 to 7, wherein the IX GSEM supplement consists of a mixture of L-alanine, L-asparagine, L-aspartic acid, L-glutamic acid, L-proline, L-serine, adenosine, guanosine, cytidine, uridine, and thymidine.

9. The medium of any one of claims 1 to 8, wherein the trace elements are selected from cupric sulfate, ferric citrate, sodium selenite, and zinc sulfate.

10. A composition comprising the medium of any one of claims 1 to 9 and cells selected from ARPE-19, Vero, MDCK, MRC-5, BHK, CEM, and LL-I cells.

11. The composition of claim 10, wherein said composition further comprises a virus.

12. The composition of claim 11, wherein said virus is a human herpesvirus.

13. The composition of claim 12, wherein said human herpesvirus is human cyto egalovirus (HCMV).

14. The composition of claim 13, wherein said HCMV is a conditional replication defective HCMV mutant.

15. The composition of claim 14, wherein the conditional replication defective

HCMV mutant has a genomic sequence of SEQ ID NO: 1.

16. A method of increasing virus production in cells, said method comprising culturing the composition of any of claims 10 to 15, wherein said composition provides increased production of the virus by at least 10% relative to the production of virus grown in a composition that does not contain the supplements of claim 1.