Non-tumorigenic MDCK cell line for propagating viruses
The present invention provides novel MDCK-derived adherent non-tumorigenic cell lines that can be grown in the presence or absence of serum. The cell lines of the present invention are useful for the production of vaccine material (e.g., viruses). More specifically, the cell lines of the present invention are useful for the production of influenza viruses in general and ca/ts influenza viruses in particular. The invention further provides methods and media formulations for the adaptation and cultivation of MDCK cells such that they remain non-tumorigenic. Additionally, the present invention provides methods for the production of vaccine material (e.g., influenza virus) in the novel cell lines of the invention.
1. A cell culture composition comprising non-tumorigenic and adherent Madin Darby Canine Kidney (MDCK) cells that have been adapted to grow in a serum-free medium comprising Taub's medium, lipids and wheat hydrolysate, which Taub's medium comprises a 50:50 mixture of DMEM and Ham's F12, 4.5 grams per liter glucose, 4mM L-glutamine, 5 micrograms per milliliter insulin, 5 micrograms per milliliter transferrin, 5picomolar triiodothyronine, 50 nanomolar hydrocortisone, 25 nanograms per milliliter prostaglandin El and 10 nanomolar sodium selenite.
2. The cell culture composition of claim 1 , wherein the MDCK cells have been passaged at least 20 passages.
3. The cell culture composition of claim 1 , wherein the MDCK cells are from the cell line MDCK-S deposited as ATCC Accession No. PTA-6500 or are from the cell line deposited as ATCC Accession No. CCL34.
4. The cell culture composition of claim 1 , wherein the MDCK cells are from the cell line MDCK-SF103 deposited as ATCC Accession No. PTA-6503.
5. The cell culture composition of claim 1 , wherein the serum-free medium is MediV SF103.
6. The cell culture composition of claim 1 , wherein the serum-free medium further comprises epidermal growth factor (EGF).
7. The cell culture composition of claim 6 , wherein the MDCK cells are from the cell line MDCK-S deposited as ATCC Accession No. PTA-6500 or are from the cell line deposited as ATCC Accession No. CCL34.
8. The cell culture composition of claim 6 , wherein the MDCK cells are from the cell line MDCK-SF103 deposited as ATCC Accession No. PTA-6503.
9. The cell culture composition of claim 6 , wherein the serum-free medium is MediV SF104 or MediV SF105.
10. The cell culture composition of claim 6 , wherein the serum-free medium further comprises tropolone and lacks transferin.
11. The cell culture composition of claim 10 , wherein the MDCK cells are from the cell line MDCK-S deposited as ATCC Accession No. PTA-6500 or are from the cell line deposited as ATCC Accession No. CCL34.
12. The cell culture composition of claim 10 , wherein the MDCK cells are from the cell line MDCK-SF103 deposited as ATCC Accession No. PTA-6503.
13. The cell culture composition of claim 10 , wherein the serum-free medium is MediV SF105.
14. The cell culture composition of claim 2 , wherein the MDCK cells have been passaged at least 30 passages.
15. The cell culture composition of claim 14 , wherein the MDCK cells have been passaged at least 40 passages.
16. The cell culture composition of claim 15 , wherein the MDCK cells have been passaged at least 50 passages.
17. The cell culture composition of claim 16 , wherein the MDCK cells have been passaged at least 60 passages.
18. An adherent, non-tumorigenic Madin Darby Canine Kidney (MDCK) cell line that can be cultivated in serum-free media and that can be infected by influenza viruses, which is prepared by a process comprising:
(a) adapting MDCK cells to grow in a serum-free medium comprising Taub's medium, lipids and wheat hydrolysate, which Taub's medium comprises a 50:50mixture of DMEM and Ham's F12, 4.5 grams per liter glucose, 4 mM L-glutamine, 5micrograms per milliliter insulin, 5 micrograms per milliliter transferrin, 5 picomolar triiodothyronine, 50 nanomolar hydrocortisone, 25 nanograms per milliliter prostaglandin E1and 10 nanomolar sodium selenite; and
(b) establishing a cell bank.
19. The adherent, non-tumorigenic MDCK cell line of claim 18 , wherein the MDCK cells are from the cell line MDCK-S deposited as ATCC Accession No. PTA-6500 or are from the cell line deposited as ATCC Accession No. CCL34.
20. The adherent, non-tumorigenic MDCK cell line of claim 18 , wherein the MDCK cells are from the cell line MDCK-SF103 deposited as ATCC Accession No. PTA-6503.
21. The adherent, non-tumorigenic MDCK cell line of claim 18 , wherein the serum-free medium is MediV SF103.
22. The adherent, non-tumorigenic MDCK cell line of claim 18 , wherein the serum-free medium further comprises epidermal growth factor (EGF).
23. The adherent, non-tumorigenic MDCK cell line of claim 22 , wherein the MDCK cells are from the cell line MDCK-S deposited as ATCC Accession No. PTA-6500 or are from the cell line deposited as ATCC Accession No. CCL34.
24. The adherent, non-tumorigenic MDCK cell line of claim 22 , wherein the MDCK cells are from the cell line MDCK-SF103 deposited as ATCC Accession No. PTA-6503.
25. The adherent, non-tumorigenic MDCK cell line of claim 22 , wherein the serum-free medium is MediV SF104 or MediV SF105.
26. The adherent, non-tumorigenic MDCK cell line of claim 22 , wherein the serum-free medium further comprises tropolone and lacks transferin.
27. The adherent, non-tumorigenic MDCK cell line of claim 26 , wherein the MDCK cells are from the cell line MDCK-S deposited as ATCC Accession No. PTA-6500 or are from the cell line deposited as ATCC Accession No. CCL34.
28. The adherent, non-tumorigenic MDCK cell line of claim 26 , wherein the MDCK cells are from the cell line MDCK-SF103 deposited as ATCC Accession No. PTA-6503.
29. The adherent, non-tumorigenic MDCK cell line of claim 26 , wherein the serum-free medium is MediV SF105.
30. The adherent, non-tumorigenic MDCK cell line of claim 18 , wherein the MDCK cells have been passaged at least 20 passages.
31. The adherent, non-tumorigenic MDCK cell line of claim 30 , wherein the MDCK cells have been passaged at least 30 passages.
32. The adherent, non-tumorigenic MDCK cell line of claim 31 , wherein the MDCK cells have been passaged at least 40 passages.
33. The adherent, non-tumorigenic MDCK cell line of claim 32 , wherein the MDCK cells have been passaged at least 50 passages.
34. The adherent, non-tumorigenic MDCK cell line of claim 33 , wherein the MDCK cells have been passaged at least 60 passages.
35. The adherent, non-tumorigenic MDCK cell line of claim 22 , wherein the serum-free medium is MediV SF104.
36. The cell culture composition of claim 6 , wherein the serum-free medium is MediV SF104.