Functionalization of plant tissues for human cell expansion

Patent metadata

Field	Value
Publication number	US-12280177-B2
Application number	US-202117388652-A
Country	US
Kind code	B2
Filing date	Jul 29, 2021
Priority date	Mar 14, 2016
Publication date	Apr 22, 2025
Grant date	Apr 22, 2025

Abstract

Decellularized plant tissues and the use of these plant tissues as scaffolds are disclosed herein. Particularly, decellularized plant tissues are functionalized such to allow for human cell adhesion, thereby allowing for their use as scaffolds for human cells. These scaffolds can then be used in a number of applications/markets, including as research tools for tissue engineering, regenerative medicine, and basic cellular biology.

First claim

The invention claimed is:

1. A plant scaffold comprising a decellularized plant tissue comprising a plant adhesion molecule conjugated to a cell adhesion peptide, wherein the plant scaffold has a perfusable structure.

2. The plant scaffold as set forth in claim 1 , wherein the decellularized plant tissue is selected from the group consisting of leaf tissue, stem tissue, root tissue, and combinations thereof.

3. The plant scaffold as set forth in claim 1 , wherein the decellularized plant tissue is derived from a plant selected from the group consisting of spinach, sweet wormwood, parsley, vanilla, peanut, and combinations thereof.

4. The plant scaffold as set forth in claim 1 , wherein the plant adhesion molecule is selected from the group consisting of a dopamine-containing compound, a polyphenol and combinations thereof.

5. The plant scaffold as set forth in claim 1 , wherein the cell adhesion peptide is selected from the group consisting of RGD, RGDS (SEQ ID NO:7), CRGDS (SEQ ID NO:8), CRGDSP (SEQ ID NO:9), CPHSRNSGSGSGSGSGRGD (SEQ ID NO:29), Acetylated-GCYGRGDSPG (SEQ ID NO:30), CRDGS (SEQ ID NO:31), cyclic RGD {Fd}C (SEQ ID NO: 32), and RGDSP (SEQ ID NO:43).

6. The plant scaffold as set forth in claim 1 , further comprising a mineral layer, wherein the mineral layer comprises a calcium to phosphate ratio of from about 2.5:1 to about 1:1.

7. The plant scaffold as set forth in claim 1 , further comprising a mineral layer, wherein the mineral layer further comprises a biomolecule selected from the group consisting of nucleic acids, proteins, peptides, growth factors, proteoglycans, and combinations thereof.

8. The plant scaffold of claim 1 , further comprising a cell.

9. The plant scaffold of claim 8 , wherein the cell is a mesenchymal stem cell.

10. A method for preparing a plant scaffold for tissue engineering, the method comprising:
decellularizing a plant tissue to provide a plant scaffold having a perfusable structure; and
contacting the decellularized plant tissue with a plant adhesion molecule conjugated to a cell adhesion peptide.

11. The method as set forth in claim 10 , wherein the decellularizing the plant tissue comprises perfusing the plant tissue with at least one detergent.

12. The method as set forth in claim 10 , wherein the plant adhesion molecule is selected from the group consisting of a dopamine-containing compound, a polyphenol and combinations thereof.

13. The method as set forth in claim 10 , wherein the cell adhesion peptide is selected from the group consisting of RGD, RGDS (SEQ ID NO:7), CRGDS (SEQ ID NO:8), CRGDSP (SEQ ID NO:9), GWGGRGDSP (SEQ ID NO:11), CPHSRNSGSGSGSGSGRGD (SEQ ID NO:29), Acetylated-GCYGRGDSPG (SEQ ID NO:30), CRDGS (SEQ ID NO:31), cyclic RGD {Fd}C (SEQ ID NO:32), and RGDSP (SEQ ID NO:43).

14. The method as set forth in claim 10 , wherein the plant scaffold supports vasculature above about 200 μm diffusion limit.

15. A cell culture method comprising contacting a cell with the plant scaffold as set forth in claim 1 .

Assignees

• Wisconsin Alumni Res Found
• Arkansas State University—Jonesboro
• Worcester Polytech Inst

Inventors

• Murphy William L
• Fontana Gianluca
• Gershlak Joshua
• Gaudette Glenn
• Weathers Pam
• Dominko Tanja
• Rolle Marsha
• Hernandez Sarah
• Cramer Carol
• Medina-Bolivar Luis Fabircio
• Binder Bernard

Classifications

• C12N5/04

  Plant cells or tissues {(culture media C12N5/0025)} · CPC title

• A61L2300/414

  Growth factors · CPC title

• A61L27/54

  Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)} · CPC title

• A61L27/227

  Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24 · CPC title

• A61L27/12

  Phosphorus-containing materials, e.g. apatite · CPC title