Apparatus and method for electroporation of biological samples
The present invention relates to methods and apparatus for the encapsulation of biologically-active substances in various cell populations. More particularly, the present invention relates to a method and apparatus for the encapsulation of biologically-active substances in various cell populations in blood by electroporation to achieve therapeutically desirable changes in the physical characteristics of the various cell populations in blood.
1. A method of producing an infectious vector comprising:
a) transfecting a cell by flow electroporation with:
i) at least one polynucleotide molecule comprising one or more viral genes, wherein the polynucleotide lacks a functional packaging signal; and
ii) at least one polynucleotide molecule comprising one or more transgenes and a viral packaging signal;
b) culturing the transfected cell; and
c) harvesting the infectious vector.
2. The method of claim 1 , wherein the infectious vector is a retroviral vector.
3. The method of claim 1 , wherein the infectious vector is a lentiviral vector.
4. The method of claim 1 , wherein at least one polynucleotide molecule is a DNA molecule.
5. The method of claim 1 , wherein at least one polynucleotide molecule is a RNA molecule.
6. The method of claim 1 , wherein the cells are transfected with at least three polynucleotide molecules.
7. The method of claim 1 , wherein the cells are adherent cells.
8. The method of claim 1 , wherein the cells are suspension cells.
9. The method of claim 1 , wherein the cells are mammalian cells.
10. The method of claim 9 , wherein the cells are human cells.
11. The method of claim 9 , wherein the cells are mouse cells.
12. The method of claim 9 , wherein the cells are hamster cells.
13. A method of producing a retroviral vector or a lentiviral vector comprising:
a) transfecting a cell by electroporation with:
i) at least one polynucleotide molecule comprising one or more viral genes, wherein the polynucleotide lacks a functional packaging signal; and
ii) at least one polynucleotide molecule comprising one or more transgenes and a viral packaging signal;
b) culturing the transfected cell; and
c) harvesting the retroviral vector or the lentiviral vector.
14. The method of claim 13 , wherein at least one polynucleotide molecule is a DNA molecule.
15. The method of claim 13 , wherein at least one polynucleotide molecule is a RNA molecule.
16. The method of claim 13 , wherein the cells are transfected with at least three polynucleotide molecules.
17. The method of claim 13 , wherein the cell is an adherent cell.
18. The method of claim 13 , wherein the cell is a suspension cell.
19. The method of claim 13 , wherein the electroporation is flow electroporation.
20. The method of claim 13 , wherein the cells are mammalian cells.
21. The method of claim 20 , wherein the cells are human cells.
22. The method of claim 21 , wherein the cells are mouse cells.
23. The method of claim 21 , wherein the cells are hamster cells.