Magneto-biolistic methods

• US 5,753,477 A
• Filed: 03/19/1996
• Issued: 05/19/1998
• Est. Priority Date: 03/19/1996
• Status: Expired due to Term

• Current Assignee: University Technology Corporation
• Original Assignee: University Technology Corporation
• Inventors: Chan, Daniel C.F.
• Primary Examiner(s): Low, Christopher S.F.
• Application Number:
  US08/617,685
• Time in Patent Office: 791 Days
• Field of Search: 435/320.1, 435/172.3, 435/172.1, 435/252.3, 435/252.33, 435/325, 435/348, 435/349, 435/351, 435/352, 435/410-431, 514/44, 935/52, 935/53, 935/54, 935/55, 935/56, 935/57, 800/2, 800/205, 800/220, 800/230, 800/250
• US Class Current: 435/455
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 15/895 using biolistic methods

Abstract

The present invention discloses methods to transfect cells, comprising applying a strong magnetic field in pulses so as to affect a plurality of substance-carrying magnetic microparticles, the complexes being in physical proximity to a plurality of cells such that when the magnetic field is applied, the magnetic microparticles are pulled into the nuclei and/or cytoplasm of the cells.

Claims

1. A method for delivery of a composition to cells in vitro, said composition comprising a plurality of substance-carrying superparamagnetic microparticles, comprising:

applying a magnetic field in a least two pulses to said composition and cells, wherein said magnetic field is 0.5-50 Teslas in strength, 0.001-200 milliseconds in duration, and insufficient to heat-kill said cells, wherein said magnetic field is applied so as to achieve penetration of the cell membrane by said substance-carrying superparamagnetic microparticles, and said cells are maintainable in viable culture post-delivery.

2. A method of claim 1, wherein the substance is complexed to the superparamagnetic microparticle via a biopolymer coating on the superparamagnetic microparticle.

3. A method of claim 1, wherein the substance is complexed to superparamagnetic microparticle via a lipid coating on the superparamagnetic microparticle.

4. A method of claim 3, wherein the substance is nucleic acid.

5. A method of claim 1, wherein the substance is nucleic acid.

6. A method of claim 1, wherein the cells are mammalian cells.

7. A method of claim 6, wherein the mammalian cells are human cells.

8. A method of claim 7, wherein the human cells are selected from the group consisting of;

brain cells;

mammary gland cells;

peripheral blood lymphocyte cells;

skin cells;

liver cells;

lung cells;

pancreatic cells;

kidney cells;

heart cells;

prostate cells;

ovary cells and spleen cells.

9. A method of claim 2, wherein the substance is a pharmaceutical compound.

10. A method of claim 9, wherein the cells are human cells.

11. A method of claim 10, wherein the human cells are selected from the group consisting of;

brain cells;

mammary gland cells;

peripheral blood lymphocyte cells;

skin cells;

liver cells;

lung cells;

pancreatic cells;

kidney cells;

heart cells;

prostate cells;

ovary cells and spleen cells.

23. A method of claim 1, wherein the cell type transfected is a plant cell.

24. A method of claim 23, wherein the plant cell type transfected is a cereal grain.

25. A method of claim 24, wherein the cereal grain cell type transfected is barley.

12. A method for delivery of a composition to cell in vitro, said composition comprising a plurality of substance-carrying ferromagnetic microparticles, comprising:

applying a magnetic field in a least two pulses to said composition, wherein said magnetic field is 0.5-50 Teslas in strength, 0.001-200 milliseconds in duration, and insufficient to heat-kill said cells, wherein said magnetic field is applied so as to achieve penetration of the cell membrane by said substance-carrying microparticles, and said cells are maintainable in viable culture post-delivery.

13. A method of claim 12, wherein the substance is complexed to the superparamagnetic microparticle via a biopolymer coating on the superparamagnetic microparticle.

14. A method of claim 12, wherein the substance is complexed to the superparamagnetic microparticle via a lipid coating on the superparamagnetic microparticle.

15. A method of claim 14, wherein the substance is a nucleic acid.

16. A method of claim 12, wherein the substance is a nucleic acid.

17. A method of claim 12, wherein the cells are mammalian cells.

18. A method of claim 17, wherein the mammalian cells are human cells.

19. A method of claim 18, wherein the human cells are selected from the group consisting of;

brain cells;

mammary gland cells;

peripheral blood lymphocyte cells;

skin cells;

liver cells;

lung cells;

pancreatic cells;

kidney cells;

heart cells;

prostate cells;

ovary cells and spleen cells.

20. A method of claim 13, wherein the substance is a pharmaceutical compound.

21. A method of claim 20, wherein the cells are human cells.

22. A method of claim 21, wherein the human cells are selected from the group consisting of;

brain cells;

mammary gland cells;

peripheral blood lymphocyte cells;

skin cells;

liver cells;

lung cells;

pancreatic cells;

kidney cells;

heart cells;

prostate cells;

ovary cells and spleen cells.

26. A method of claim 12, wherein the cell type transfected is a plant cell.

27. A method of claim 26, wherein the plant cell type transfected is a cereal grain.

28. A method of claim 27, wherein the cereal grain cell type transfected is barley.