THREE DIMENSIONAL CELL CULTURE COMPOSITIONS AND METHODS OF USE

• US 20140051168A1
• Filed: 08/08/2013
• Published: 02/20/2014
• Est. Priority Date: 08/17/2012
• Status: Active Grant

• Current Assignee: Lena Biosciences, Inc.
• Original Assignee: Lena Biosciences, Inc.
• Inventors: Vukasinovic, Jelena
• Granted Patent: US 9,334,473 B2
• Time in Patent Office: Days
• US Class Current: 435/397
• CPC Class Codes: C12N 2533/12 Glass
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Described herein is a three-dimensional cell culture scaffold composition comprising an absorbent rigid (AR) component, and in some embodiments, further comprises a gel component. The absorbent rigid component preferably comprises a glass fiber material. It is a surprising finding of the present invention that an AR component having a void volume of between approximately 70% and 95% results in a three-dimensional cell culture composition that allows for robust, high-throughput screening and high-content screening accessible tissue models with preserved cell morphology, heterogeneity of cell types and cell populations, extracellular matrix constituents, functional cell-cell and cell-extracellular matrix interactions and signaling with sufficient specificities to tissue physiology and pathology.

Claims

1. A three-dimensional cell culture scaffold composition comprising a rigid absorbent component and cells, wherein the rigid absorbent component has a void volume of between approximately 70% and 95%, which void volume comprises one or more cells.

2. The composition of claim 1, wherein the void volume further comprises a gel.

3. The composition of claim 1, wherein the rigid absorbent component comprises glass fibers.

4. The composition of the claim 3, wherein the glass fibers are borosilicate glass fibers.

5. The composition of claim 3, wherein the glass fibers have a diameter between approximately 3 and 30 μ

m.

6. The composition of claim 3, wherein the rigid absorbent component is a Millipore G041 material.

7. The composition of claim 3, wherein the rigid absorbent component comprises approximately 70 to 95% w/w glass fibers.

8. The composition of claim 1, wherein the rigid absorbent component comprises an absorbent coating.

9. The composition of claim 8, wherein the absorbent coating is poly vinyl alcohol.

10. The composition of claim 2, wherein the gel is a MATRIGEL™

.

11. The composition of claim 1, wherein the void volume is between approximately 85-95%.

12. The composition of claim 1, wherein the scaffold further comprises a gradient of one or more molecules.

13. A method of culturing or suspending cells comprising placing the cells in a three-dimensional cell culture scaffold composition, which composition comprises a rigid absorbent component and has a void volume of between approximately 70% and 95%, thereby creating a three dimensional cell culture or suspension.

14. The method of claim 13, wherein the void volume further comprises a gel.

15. The method of claim 13, wherein the rigid absorbent component comprises glass fibers.

16. The method of claim 15, wherein the glass fibers are borosilicate glass fibers.

17. The method of claim 15, wherein the glass fibers have a diameter between approximately 3 and 30 μ

m.

18. The method of claim 15, wherein the rigid absorbent component is a Millipore G041 material.

19. The method of claim 15, wherein the rigid absorbent component comprises approximately 70 to 95% w/w glass fibers.

20. The method of claim 13, wherein the rigid absorbent component comprises an absorbent coating.

21. The method of claim 20, wherein the absorbent coating is poly vinyl alcohol.

22. The method of claim 14, wherein the gel is a MATRIGEL™

.

23. The method of claim 13, wherein the void volume is between approximately 85-95%.

24. The method of claim 13, wherein the rigid absorbent component comprises a poly-D-lysine coating.

25. The method of claim 23, further comprising a high content or high throughput screening of the three-dimensional cell culture.

26. The method of claim 23, further comprising placing two or more of the rigid absorbent components vertically or horizontally adjacent to one another.

27. A method of adding one or more molecules comprising placing the one or more molecules in a rigid absorbent component which has a void volume of between approximately 70% and 95%, thereby creating a uniform or non-uniform concentration of the one or more molecules within the rigid absorbent component.

28. The method of claim 27, wherein the rigid absorbent component further comprises a coating, a gel, one or more cells, a coating and a gel, coating and one or more cells, gel and one or more cells, or a coating, a gel and one or more cells.

29. The method of claim 27, wherein different molecules or different concentrations of the same molecules are placed at different locations so as to form a gradient.