DEVICES, SYSTEMS, AND METHODS FOR THE FABRICATION OF TISSUE UTILIZING UV CROSS-LINKING

• US 20140093932A1
• Filed: 03/11/2013
• Published: 04/03/2014
• Est. Priority Date: 04/20/2012
• Status: Active Grant

• Current Assignee: Organovo Inc. (Organovo Holdings, Inc.)
• Original Assignee: Organovo Inc. (Organovo Holdings, Inc.)
• Inventors: Murphy, Keith, Dorfman, Scott, Law, Richard Jin, Le, Vivian Anne
• Granted Patent: US 9,499,779 B2
• Time in Patent Office: Days
• US Class Current: 435/173.4
• CPC Class Codes: B29C 64/112 using individual droplets, e.g. from jetting heads
• CPC Class Codes: B33Y 10/00 Processes of additive manufacturing
• CPC Class Codes: B33Y 30/00 Apparatus for additive manufacturing; Details thereof or accessories therefor
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: B41J 3/407 for marking on special material
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 29/00 Means for introduction, extraction or recirculation of materials, e.g. pumps pumps per se F04B
• CPC Class Codes: C12M 33/00 Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture

Abstract

Described herein are bioprinters comprising: one or more printer heads, wherein a printer head comprises a means for receiving and holding at least one cartridge, and wherein said cartridge comprises contents selected from one or more of: bio-ink, and support material; a UV light module for optionally exposing the contents of at least one cartridge to UV light; a means for calibrating the position of at least one cartridge; and a means for dispensing the contents of at least one cartridge. Also described herein are methods of using and bioprinting cartridges for such bioprinters.

Claims

1. A bioprinter comprising:

a. one or more printer heads, wherein a printer head comprises a means for receiving and holding at least one cartridge, and wherein said cartridge comprises contents selected from one or more of;

bio-ink and support material;

b. a UV light module for optionally exposing the contents of at least one cartridge to UV light; and

c. a means for dispensing the contents of at least one cartridge.

2. The bioprinter of claim 1, wherein the means for dispensing the contents of at least one cartridge is application of a piston, application of pressure, application of compressed gas, hydraulics, or a combination thereof.

3. The bioprinter of claim 1, wherein the bio-ink or support material comprises one or more UV cross-linkable materials.

4. The bioprinter of claim 1, wherein the UV light module exposes the contents of at least one cartridge to UV light prior to dispensing.

5. The bioprinter of claim 1, wherein the UV light module is tunable with respect to wavelength, intensity, exposure time, or any combination thereof.

6. The bioprinter of claim 5, wherein the UV light module comprises one or more optionally engaged attenuation filters, wherein the attenuation filters lower UV intensity when engaged.

7. The bioprinter of claim 1, further comprising a means for applying a wetting agent to one or more of:

a. a printer stage;

b. a receiving surface;

c. a deposition orifice;

d. the bio-ink;

e. the support material; and

f. a printed construct;

wherein the wetting agent is applied at one or more time points selected from;

before the bio-ink or support material is dispensed by the bioprinter, substantially concurrently with dispensing, and after the bio-ink or support material is dispensed by the bioprinter.

8. A method of fabricating a tissue construct comprising the steps of:

a. depositing bio-ink onto a support, wherein the bio-ink comprises a UV cross-linkable material and has been exposed to UV light to partially or completely cross-link the UV cross-linkable material to form a semi-solid or solid body; and

b. incubating the bio-ink to allow it to cohere and to form a tissue, wherein said incubation has a duration of about 1 hour to about 10 days.

9. The method of claim 8, further comprising the step of preparing a liquid bio-ink solution comprising a UV cross-linkable material and cells.

10. The method of claim 9, further comprising the step of exposing a liquid bio-ink solution comprising a UV cross-linkable material and cells to UV light to partially or completely cross-link the UV cross-linkable material to form a semi-solid or solid body.

11. The method of claim 8, further comprising the step of depositing bio-ink onto a support, wherein the bio-ink comprises a UV cross-linkable material and has not been exposed to UV light.

12. The method of claim 8, wherein the UV cross-linkable material is a hydrogel.

13. The method of claim 8, wherein the bio-ink comprises mammalian cells.

14. The method of claim 13, wherein the bio-ink comprises differentiated, mammalian cells.

15. The method of claim 13, wherein the bio-ink comprises non-differentiated, mammalian cells.

16. The method of claim 8, wherein the bio-ink is deposited by extrusion from a bioprinter.

17. A bioprinter cartridge, wherein the contents of the cartridge comprises a UV cross-linkable material and optionally cells.

18. The bioprinter cartridge of claim 17, wherein the UV cross-linkable material is partially or completely cross-linked.

19. The bioprinter cartridge of claim 17, wherein the UV cross-linkable material is a hydrogel.

20. The bioprinter cartridge of claim 17, wherein the cartridge comprises a glass-walled reservoir.

21. The bioprinter cartridge of claim 17, wherein the cartridge is permeable to UV light.

22. A UV light module for a bioprinter comprising a UV light source that optionally exposes the contents of at least one bioprinter cartridge to UV light.

23. The UV light module of claim 22, wherein the UV light module is reversibly attachable to a bioprinter.

24. The UV light module of claim 22, wherein the UV light module is tunable with respect to wavelength, intensity, exposure time, or any combination thereof.

25. The UV light module of claim 24, further comprising one or more optionally engaged attenuation filters, wherein the attenuation filters lower UV intensity when engaged.

26. A tissue construct comprising a multicellular body on a support substrate, wherein the multicellular body comprises a UV cross-linkable material and cells, wherein the multicellular body was deposited by a bioprinter, provided that the multicellular body was exposed to UV light to partially or completely cross-link the UV cross-linkable material to form a semi-solid or solid body.

27. The tissue construct of claim 26, wherein the construct comprises a plurality of multicellular bodies.

28. The tissue construct of claim 26, wherein the UV cross-linkable material is a hydrogel.

29. The tissue construct of claim 26, wherein the multicellular body comprises differentiated, mammalian cells.

30. The tissue construct of claim 26, wherein the multicellular body comprises non-differentiated, mammalian cells.