Bio-ink structures and methods of producing the same

Patent metadata

Field	Value
Publication number	US-12404413-B2
Application number	US-201816153501-A
Country	US
Kind code	B2
Filing date	Oct 5, 2018
Priority date	Oct 5, 2018
Publication date	Sep 2, 2025
Grant date	Sep 2, 2025

Abstract

Bio-ink comprising freeze-dried cells, methods of making a living structure from a bio-ink material of freeze-dried cells, and methods of using the living structure for biosensing, tissue regeneration, environment sensing, drug discovery, catalysis, and/or clinical implementation are described herein.

First claim

We claim:

1. A bio-ink comprising:
a first filler component comprising freeze-dried cells, wherein the first filler component of freeze-dried cells has a cell density of at least about 10 vol % or more of the total volume of the bio-ink, and
a second filler component comprising nanocellulose crystalline powder,
wherein the bio-ink is configured to be used for bioprinting three-dimensional structures including living cells.

2. The bio-ink of claim 1 , wherein the first filler component of freeze-dried cells are microbes, wherein the microbes are selected from the group consisting of bacteria, algae, fungi, protozoa, and a mixture thereof.

3. The bio-ink of claim 1 , wherein the first filler component of freeze-dried cells has a cell density of at least about 20 vol % or more of the total volume of the bio-ink.

4. The bio-ink of claim 1 , wherein the first filler component of freeze-dried cells has a cell density of at least about 40 vol % or more of the total volume of the bio-ink.

5. The bio-ink of claim 1 , wherein the first filler component of freeze-dried cells has a cell density of at least about 60 vol % or more of the total volume of the bio-ink.

6. The bio-ink of claim 1 , wherein the first filler component of freeze-dried cells has a cell density of at least about 80 vol % or more of the total volume of the bio-ink.

7. A bio-ink comprising:
a first filler component comprising freeze-dried cells, wherein the first filler component of freeze-dried cells has a cell density of at least about 10 wt % of the total weight of the bio-ink and
a second filler comprising nanocellulose crystalline powder,
wherein the bio-ink is configured to be used for bioprinting three-dimensional structures including living cells.

8. The bio-ink of claim 7 , wherein the first filler component of freeze-dried cells are microbes, and wherein the microbes are selected from the group consisting of bacteria, algae, fungi, protozoa, and a mixture thereof.

9. The bio-ink of claim 7 , wherein the first filler component of freeze-dried cells has a cell density of at least about 20 wt % or more of the total weight of the bio-ink.

10. The bio-ink of claim 7 , wherein the first filler component of freeze-dried cells has a cell density of at least about 40 wt % or more of the total weight of the bio-ink.

11. The bio-ink of claim 7 , wherein the first filler component of freeze-dried cells has a cell density of at least about 60 wt % or more of the total weight of the bio-ink.

12. The bio-ink of claim 7 , wherein the first filler component of freeze-dried cells has a cell density of at least about 80 wt % or more of the total weight of the bio-ink.

13. A bio-ink comprising:
a first filler component comprising freeze-dried cells, wherein the first filler component of freeze-dried cells has a cell density of at least about 10 vol % or more of the total volume of the bio-ink,
a second filler component selected from the group consisting of nanocellulose, cellulose nanocrystals (CNC), cellulose nanofibrils (CNF), and nanocellulose crystalline powder,
wherein the bio-ink is configured to be used for bioprinting three-dimensional structures including living cells, and

a binder comprising an oligomer, monomer, or a combination thereof, wherein the binder is configured to form a polymer when the bio-ink is used for bio-printing the three-dimensional structures including living cells.

14. The bio-ink of claim 13 , wherein the binder comprises styrene, methacrylate, vinyl alcohol, polyisobutylene, glycerol, polypropylene, or polyethylene glycol dimethacrylate.

15. The bio-ink of claim 13 , further comprising a photo-initiator configured to initiate photopolymerization of the binder upon irradiation with light.

16. The bio-ink of claim 15 , wherein the photo-initiator comprises 1-hydroxycyclohexyl phenyl ketone, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-1-[4-(-methylthio)phenyl]-2-4-morpholinyl)-1-propanone, hydroxyacetophenone, phosphineoxide, benzophenone, or lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP).

17. The bio-ink of claim 13 , wherein the first filler component of freeze-dried cells are microbes, wherein the microbes are selected from the group consisting of bacteria, algae, fungi, protozoa, and a mixture thereof.

18. The bio-ink of claim 13 , wherein the first filler component of freeze-dried cells has a cell density of at least about 20 vol % or more of the total volume of the bio-ink.

19. The bio-ink of claim 13 , wherein the first filler component of freeze-dried cells has a cell density of at least about 40 vol % or more of the total volume of the bio-ink.

20. The bio-ink of claim 13 , wherein the first filler component of freeze-dried cells has a cell density of at least about 60 vol % or more of the total volume of the bio-ink.

Assignees

• L Livermore Nat Security Llc

Inventors

• Qian Fang
• Baker Sarah
• Deotte Joshua R
• Duoss Eric
• Knipe Jennifer Marie
• Ruelas Samantha
• Spadaccini Christopher
• Stolaroff Joshuah K
• Zhu Cheng

Classifications

• B33Y70/00

  Materials specially adapted for additive manufacturing · CPC title

• B29C64/106

  using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material · CPC title

• B33Y10/00

  Processes of additive manufacturing · CPC title

• B29K2105/16

  Fillers · CPC title

• C12P7/06

  Ethanol, i.e. non-beverage · CPC title