Method for producing continuous cell lines
View Patent ↗The present invention relates to a method for production of continuous cell lines comprising providing living cells of an animal or a human, irradiating said cells with UV light, proliferating said cells and selecting multiplying cells as cells of a continuous cell line.
1. A method for production of a continuous avian cell line comprising irradiating cultured isolated primary avian cells with a UV light having a wavelength between 200 nm to 300 nm and with a dose from about 20 mJ/cm 2 to about 105 mJ/cm 2 , wherein the irradiation time is from 30 seconds to 4 minutes, and selecting cells capable to proliferate after at least 20 passages as cells of a continuous avian cell line.
2. The method of claim 1 wherein said UV light dosage is at least 50 mJ/cm 2 .
3. The method of claim 1 wherein selecting said cells comprises at least 40 passages.
4. The method of claim 1 wherein said cells are attached to a surface or are in suspension.
5. The method of claim 1 wherein said cells are cells of an embryo.
6. The method of claim 1 wherein said cells are a mixed culture of more than one type of tissue.
7. The method of claim 1 wherein said cells are in a monolayer.
8. The method of claim 1 further comprising infecting said cells with a virus under conditions which permit virus proliferation.
9. The method of claim 8 wherein said virus is selected from baculovirus, poxvirus, adenovirus, papovavirus, parvovirus, hepadnavirus, coronavirus, flavivirus, togavirus, astrovirus, picornavirus, retrovirus, orthomyxovirus, filovirus, paramyxovi-rus, rhabdovirus, arenavirus, and bunyavirus.
10. The method of claim 8 wherein said virus is selected from modified vaccinia virus Ankara (MVA), tick-borne encephalitis (TBE) virus, Yellow fever virus, West Nile virus, New Caledonia virus or an influenza virus.
11. The method of claim 1 further comprising transfecting said cells with a nucleic acid encoding a gene product, growing said cells under conditions which permit production of said gene product.
12. The method of claim 1 wherein said cells of said cell line are adapted for growth in serum free media.
13. The method of claim 12 wherein said medium is selected from DMEM/HAM's F12, RPMI, MEM, BME, Waymouth's medium, an oligopeptide free medium, a chemically defined medium or a combination thereof.
14. The method of claim 1 wherein said cells of said cell lines are selected for being non-tumorigenic and/or non-carcinogenic.
15. The method of claim 1 , wherein the dose is from about 70 mJ/cm 2 to about 105 mJ/cm 2 .
16. The method of claim 8 , further comprising collecting said virus.
17. The method of claim 11 , further comprising collecting said gene product.