Three-dimensional discontinuous entity for cell culturing

Patent metadata

Field	Value
Publication number	US-9593304-B2
Application number	US-201314430833-A
Country	US
Kind code	B2
Filing date	Sep 24, 2013
Priority date	Sep 25, 2012
Publication date	Mar 14, 2017
Grant date	Mar 14, 2017

Abstract

The present invention is related to methods and materials for culturing and transporting stem cells in a three-dimensional culture. The materials comprise cellulose nanofibrils in a form of three-dimensional discontinuous entities in a biocompatible hydrogel.

First claim

The invention claimed is:

1. A three-dimensional discontinuous entity for culturing of cells comprising
a. an aqueous medium comprising a mixture of a first and second aqueous medium; and
b. hydrogel bodies comprising cellulose nanofibrils and/or derivatives thereof suspended in the aqueous medium, the cellulose nanofibrils being obtained from plant-derived, mechanically disintegrated cellulose, wherein the cellulose nanofibrils and/or derivatives thereof are suspended in the aqueous medium, wherein the hydrogel bodies have discontinuous structures, are separate or interconnected, and have different sizes and forms.

2. The three-dimensional discontinuous entity according to claim 1 , wherein the ratio of total volume of the hydrogel bodies to total volume of the three-dimensional discontinuous entity is 10%-99% (v/v).

3. The three-dimensional discontinuous entity according to claim 1 , wherein the hydrogel bodies are interconnected.

4. The three-dimensional discontinuous entity according to claim 1 , wherein a yield stress of the three-dimensional discontinuous entity is lower than a yield stress of the corresponding continuous hydrogel in the same conditions.

5. The three-dimensional discontinuous entity according to claim 1 , wherein a yield stress of the three-dimensional discontinuous entity is 1-95% of a yield stress of the corresponding continuous hydrogel in the same conditions.

6. The three-dimensional discontinuous entity according to claim 1 , wherein the diameter of the cellulose nanofibrils is less than 1 μm.

7. The three-dimensional discontinuous entity according to claim 1 , wherein the cellulose nanofibrils comprise native ion-exchanged, chemically modified or physically modified derivatives of cellulose nanofibrils or nanofibril bundles.

8. The three-dimensional discontinuous entity according to claim 1 , wherein the aqueous medium is a cell culture medium comprising at least one nutrient source and at least one component required for sustaining undifferentiated, differentiating or differentiated cell growth.

9. A cell culture matrix comprising a cell or cells and/or a tissue or tissues and a three-dimensional discontinuous entity according to claim 1 , wherein the cell or cells and/or tissue or tissues are present at least partially embedded in said entity in a three-dimensional or two-dimensional arrangement.

10. The cell culture matrix according to claim 9 , wherein the cells are differentiated or undifferentiated mammalian cells.

11. The cell culture matrix according to claim 9 , wherein the cells are undifferentiated stem cells.

12. The cell culture matrix according to claim 9 , wherein the cells are embryonic stem cells or induced pluripotent stem cells.

13. The cell culture matrix according to claim 9 , wherein the cells are human cells or non-human cells.

14. The cell culture matrix according to claim 13 wherein the cells are non-human embryonic stem cells or induced pluripotent stem cells.

15. A kit comprising
a first container comprising the three-dimensional discontinuous entity according to claim 1 or the three-dimensional discontinuous entity according to claim 1 in dehydrated form such as dry powder, concentrated granulate, or concentrated hydrogel body, and
a second container comprising cellulase.

16. The three-dimensional discontinuous entity of claim 1 further comprising one or more components selected from the group consisting of a cell culture medium, an extra cellular matrix component, a serum, growth factor, a protein, an antibiotic, and a preservative.

17. The three-dimensional discontinuous entity of claim 1 , wherein the discontinuous structure of the hydrogel bodies is a homogeneous structure broken by mixing.

18. The three-dimensional discontinuous entity of claim 1 , wherein the hydrogel bodies are interconnected hydrogel bodies having aqueous cavities positioned therebetween.

19. The three-dimensional discontinuous entity of claim 1 , wherein the hydrogel bodies are loosely connected and floating within the mixture.

20. The three-dimensional discontinuous entity of claim 1 , further comprising cells, the hydrogel bodies being indirectly connected to one another via the cells.

21. The three-dimensional discontinuous entity of claim 1 , wherein the second aqueous medium is a cell culture medium.

22. The three-dimensional discontinuous entity of claim 1 , wherein proteases are undetectable in the three-dimensional discontinuous entity.

23. The three-dimensional discontinuous entity of claim 1 , wherein the cellulose nanofibrils are sterilized.

Assignees

• Upm Kymmene Corp

Inventors

• Laukkanen Antti
• Lou Yan-Ru
• Yliperttula Marjo
• Kuisma Tytti
• Nikander Johanna
• Pere Jaakko

Classifications

• C12N5/0062

  General methods for three-dimensional culture · CPC title

• C12N5/0606

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title

• C12N5/0602 Primary

  Vertebrate cells · CPC title

• C12N5/0696

  Artificially induced pluripotent stem cells, e.g. iPS · CPC title

• C12N2533/78

  Cellulose · CPC title