Devices, systems, and methods for the fabrication of tissue utilizing UV cross-linking

• US 9,499,779 B2
• Filed: 03/11/2013
• Issued: 11/22/2016
• 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
• Primary Examiner(s): Long, Scott
• Assistant Examiner(s): NGUYEN, NGHI V
• Application Number:
  US13/794,368
• Publication Number:
  US 20140093932A1
• Time in Patent Office: 1,352 Days
• Field of Search: 435/173.4, 435/286.3, 435/177, 347/20, 424/572, 700/118
• US Class Current: 1/1
• 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 at least one printer head comprises a means for receiving and holding at least one cartridge, wherein at least one cartridge comprises a capillary tube, and wherein said capillary tube comprises a bio-ink, wherein the bio-ink comprises a solid or semi-solid composition of living cells;

b. a UV light module comprising a chamber, wherein the at least one cartridge is receivable in the chamber and the UV light module is configured to expose UV light in the chamber prior to or during extrusion of the bio-ink; and

c. a means for extruding the bio-ink of the capillary tube by the application of pressure.

2. The bioprinter of claim 1, wherein the means for extruding the contents of the capillary tube by the application of pressure, is by application of a piston, application of compressed gas, application of hydraulics, or a combination thereof.

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

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

5. The bioprinter of claim 4, wherein the UV light module comprises one or more attenuation filters, wherein the attenuation filters lower UV intensity when engaged.

6. 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; and

e. a printed construct;

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

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

7. The bioprinter of claim 1, wherein the at least one cartridge is receivable in the chamber and configured to expose UV light in the chamber during extrusion of the bio-ink.

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

a. depositing bio-ink onto a support with the bioprinter of claim 1, 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.