Inkjet gene printing

Patent metadata

Field	Value
Publication number	US-9598704-B2
Application number	US-201514697704-A
Country	US
Kind code	B2
Filing date	Apr 28, 2015
Priority date	Jun 7, 2007
Publication date	Mar 21, 2017
Grant date	Mar 21, 2017

Abstract

Provided herein are methods and apparatuses for transfecting a cell with a compound of interest by stressing the cell, e.g. with shear stress. The compound of interest may be nucleic acids, proteins, molecules, nanoparticles, drugs, etc., or any combination thereof. Methods of printing cells with an inkjet printing device are also provided, wherein at least a portion of viable cells (preferably at least 1%) are transfected with a compound of interest. Preferably, at least 25% of the cells are viable after printing. In addition, methods of forming an array of viable cells are provided wherein at least a portion of the viable printed cells (preferably at least 1%) are transfected with at least one compound of interest.

First claim

That which is claimed is:

1. A method for transfecting cells with a compound of interest, wherein at least a portion of said cells are transfected with said compound of interest, said method comprising the steps of:
providing a composition comprising said cells in the presence of said compound of interest in a liquid carrier; and then
forcing said composition through an orifice so that said cells are stressed in the presence of said compound of interest, wherein said orifice has a diameter of between one-eighth and twelve times the average diameter of said cells, and wherein said cells are stressed for a period of from 0.1 to 10 microseconds;
thereby transfecting at least a portion of said cells with said compound of interest,
wherein said forcing step is carried out by inkjet printing of said cells with an inkjet printing device.

2. The method of claim 1 , wherein said inkjet printing device is selected from the group consisting of thermal inkjet printers and piezoelectric inkjet printers.

3. The method of claim 1 , wherein said inkjet printing device comprises at least one inkjet cartridge, and wherein said inkjet cartridge is selected from the group consisting of thermal inkjet printer cartridges and piezoelectric inkjet printer cartridges.

4. A method of printing cells, wherein at least a portion of said cells are transfected with a compound of interest, said method comprising the steps of:
providing an inkjet printing device, said device comprising at least one inkjet printer cartridge;
loading a composition to be printed into said printer cartridge, such that said composition upon loading comprises said cells in the presence of said compound of interest; and
printing said composition onto a substrate, wherein said cells are printed through an orifice having a diameter of between one-eighth and twelve times the average diameter of said cells, and wherein said cells are stressed for a period of from 0.1 to 10 microseconds during said printing;
thereby transfecting at least a portion of said cells with said compound of interest.

5. The method of claim 4 , wherein said substrate of said printing step is coated with collagen.

6. The method of claim 4 , wherein said printing step is carried out by printing said composition onto a tissue substrate in vivo.

7. The method of claim 4 , wherein said at least a portion of said cells is at least 1% of cells that are viable after said printing step.

8. The method of claim 4 , wherein said at least a portion of said cells is at least 5% of cells that are viable after said printing step.

9. The method of claim 4 , wherein said at least a portion of said cells is at least 10% of cells that are viable after said printing step.

10. The method of claim 4 , wherein at least 25% of the cells in said loading step are viable after said forcing step.

11. The method of claim 4 , wherein at least 50% of the cells in said loading step are viable after said forcing step.

12. The method of claim 4 , wherein at least 75% of the cells in said loading step are viable after said forcing step.

13. The method of claim 4 , wherein said printer cartridge of said providing step comprises a plurality of nozzles, and wherein said nozzles have a diameter of from 0.01 to 100 micrometers.

14. The method of claim 4 , wherein said cells of said loading step are eukaryotic cells.

15. The method of claim 4 , wherein said cells of said loading step are prokaryotic cells.

16. The method of claim 4 , wherein said compound of interest of said loading step is selected from the group consisting of: nucleic acids, proteins, molecules, nanoparticles and drugs.

17. The method of claim 4 , wherein said compound of interest of said loading step comprises a nucleic acid.

18. The method of claim 4 , wherein said composition of said loading step is premixed.

19. The method of claim 4 , wherein said inkjet printing device of said providing step is selected from the group consisting of thermal inkjet printers and piezoelectric inkjet printers.

20. The method of claim 4 , wherein said inkjet printer cartridge of said providing step is selected from the group consisting of thermal inkjet printer cartridges and piezoelectric inkjet printer cartridges.

21. A method of forming an array of viable cells wherein at least a portion of said cells are transfected with at least one compound of interest, said method comprising the steps of:
providing an inkjet printing device, said device comprising at least one inkjet printer cartridge;
loading a composition to be printed into said printer cartridge, such that said composition upon loading comprises cells in the presence of at least one compound of interest; and
printing said composition onto a substrate in an organized pattern, wherein said cells are printed through an orifice having a diameter of between one-eighth and twelve times the average diameter of said cells, and wherein said cells are stressed for a period of from 0.1 to 10 microseconds during said printing;
thereby forming an array of viable cells wherein at least a portion of said cells are transfected with said at least one compound of interest.

22. The method of claim 21 , wherein said substrate of said printing step is coated with collagen.

23. The method of claim 21 , wherein said printing step is carried out by printing said composition onto a tissue substrate in vivo.

24. The method of claim 21 , wherein said at least a portion of said cells is at least 1% of cells that are viable after said printing step.

25. The method of claim 21 , wherein said at least a portion of said cells is at least 5% of cells that are viable after said printing step.

26. The method of claim 21 , wherein said at least a portion of said cells is at least 10% of cells that are viable after said printing step.

27. The method of claim 21 , wherein at least 25% of the cells in said loading step are viable after said printing step.

28. The method of claim 21 , wherein at least 50% of the cells in said loading step are viable after said printing step.

29. The method of claim 21 , wherein at least 75% of the cells in said loading step are viable after said printing step.

30. The method of claim 21 , wherein said printer cartridge of said providing step comprises a plurality of nozzles, and wherein said nozzles have a diameter of from 0.01 to 100 micrometers.

31. The method of claim 21 , wherein said cells of said loading step are eukaryotic cells.

32. The method of claim 21 , wherein said cells of said loading step are prokaryotic cells.

33. The method of claim 21 , wherein said compound of interest of said loading step is selected from the group consisting of: nucleic acids, proteins, molecules, nanoparticles and drugs.

34. The method of claim 21 , wherein said compound of interest of said loading step comprises a nucleic acid.

35. The method of claim 21 , wherein said composition of said loading step is premixed.

36. The method of claim 21 , wherein said inkjet printing device of said providing step is selected from the group consisting of thermal inkjet printers and piezoelectric inkjet printers.

37. The method of claim 21 , wherein said inkjet printer cartridge of said providing step is selected

Assignees

• Univ Wake Forest Health Sciences

Inventors

• Xu Tao
• Yoo James J
• Atala Anthony

Classifications

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C12N15/89

  using microinjection · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• C12N15/895 Primary

  using biolistic methods · CPC title

• A61K31/713

  Double-stranded nucleic acids or oligonucleotides · CPC title