Bio-ink structures and methods of producing the same

Patent metadata

Field	Value
Publication number	US-2020109299-A1
Application number	US-201816153501-A
Country	US
Kind code	A1
Filing date	Oct 5, 2018
Priority date	Oct 5, 2018
Publication date	Apr 9, 2020
Grant date	—

Abstract

In various embodiments, the present disclosure provides 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.

First claim

I/We claim:

1 . A bio-ink comprising a filler of freeze-dried cells.

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

3 . The bio-ink of claim 1 , wherein the filler of freeze-dried cells has a cell density of at least about 5 wt %, about 10 wt %, about 20 wt %, about 30 wt %, about 40 wt %, about 50 wt %, about 60 wt %, about 70 wt %, about 80 wt %, about 90 wt %, about 95 wt % or more of the total weight of the bio-ink or at least about 5 vol %, about 10 vol %, about 20 vol %, about 30 vol %, about 40 vol %, about 50 vol %, about 60 vol %, about 70 vol %, about 75 vol %, about 80 vol %, about 85 vol %, about 90 vol %, about 95 vol %, or more of the total volume of the bio-ink.

4 . The bio-ink of claim 1 , wherein the filler is a single filler or a dual filler.

5 . The bio-ink of claim 4 , wherein the dual filler comprises freeze-dried live cells and a dual filler component.

6 . The bio-ink of claim 5 , wherein the dual filler is cellulose selected from the group consisting of nanocellulose, cellulose nanocrystals (CNC), cellulose nanofibrils (CNF) bacterial cellulose (BC), and nanocellulose crystalline powder.

7 . The bio-ink of claim 1 , further comprising a hydrogel.

8 . A method of making a living structure comprising use of the bio-ink of claim 1 .

9 . The method of claim 8 , further comprising seeding at least one additional cell into the living structure.

10 . A living structure made from a bio-ink comprised of a filler of freeze-dried cells.

11 . The living structure of claim 10 , wherein the living structure is three dimensional (3D), a lattice, a scaffold, and/or porous.

12 . The living structure of claim 10 , wherein at least a portion of the freeze-dried cells are encapsulated in the living structure.

13 . The living structure of claim 10 , wherein the living structure maintains cell viability and/or is metabolically active for at least about 1 month, about 2 months, about 3 months, 4 months, about 6 months, about 8 months, or about 10 months.

14 . The living structure of claim 10 , wherein the living structure of freeze-dried cells has a cell density of at least about 5 wt %, about 10 wt %, about 20 wt %, about 30 wt %, about 40 wt %, about 50 wt %, about 60 wt %, about 70 wt %, about 80 wt %, about 90 wt %, about 95 wt % or more of the total weight of the bio-ink or least about 5 vol %, about 10 vol %, about 20 vol %, about 30 vol %, about 40 vol %, about 50 vol %, about 60 vol %, about 70 vol %, about 75 vol %, about 80 vol %, about 85 vol %, about 90 vol %, about 95 vol %, or more of the total volume of the bio-ink.

15 . The living structure of claim 10 , wherein the living structure has a resolution of at least about 10 μm, 20 μm, about 40 μm, about 50 μm, about 80 μm, about 100 μm, about 200 μm, about 300 μm, about 400 μm, and/or about 500 μm.

16 . The living structure of claim 10 , wherein the living structure has a tunable intercellular distance.

17 . The living structure of claim 10 , wherein the living structure exhibits high mass transport.

18 . A method of using a living structure made from a filler comprised freeze-dried cells for biosensing, tissue regeneration, environment sensing, drug discovery, catalysis, and/or clinical implementation.

19 . The method of claim 18 , wherein the living structure mimics morphology, intercellular interactions, signaling pathway activation, and/or diffusion of cells by performing cellular functions selected from the group consisting of cell proliferation, gene expression, protein expression, and responding to external stimuli.

20 . The method of claim 18 , wherein the catalysis is biocatalysis selected from the group consisting of food fermentation, biofuel production, protein synthesis, wastewater treatment, and bioremediation.

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

• C09D11/14 Primary

  based on carbohydrates · CPC title

• C09D11/03 Primary

  characterised by features other than the chemical nature of the binder · CPC title

• B29C64/106

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

• C09D11/101

  Inks specially adapted for printing processes involving curing by wave energy or particle radiation, e.g. with UV-curing following the printing · CPC title

• C09D11/30

  Inkjet printing inks · CPC title