Self-contained mold for culture and transport of 3D cellular gels

Patent metadata

Field	Value
Publication number	US-12180449-B2
Application number	US-201916284449-A
Country	US
Kind code	B2
Filing date	Feb 25, 2019
Priority date	Apr 6, 2018
Publication date	Dec 31, 2024
Grant date	Dec 31, 2024

Abstract

A mold capable of preparing a cellular or acellular construct includes a block including a three-dimensional cavity for a sample, a base, and a removable sheet inserted between the block and the base. The sheet may be coated with silicone grease prior to insertion of the sheet between the block and the base. Mechanical constraints may be included at each end of the cavity to constrain cell driven contraction and to allow for gripping the sample after the sample is removed from the mold. The sample may be prepared by partially filling the cavity with gel, filling the remaining portion of the cavity with culture media, and allowing a three-dimensional sample to culture within the cavity for a time. The sample may be transported to a bioreactor and ejected from the mold by removing the sheet and applying pressure to the mechanical constraints.

First claim

What is claimed is:

1. A system for preparing and transporting a cellular or acellular construct for controlled extraction of the construct, the system comprising:
a sample constructed within a three-dimensional cellular scaffold and culture media;
a block including a block height of at least twenty millimeters, a block width that is more than the block height, a block length that is more than the block width, a three-dimensional cavity for constructing the sample in three dimensions, wherein the block is made from a bio-inert material that does not produce an adverse effect with said sample, wherein the three-dimensional cavity comprises:
a cavity height that is identical to the block height,
a first sub-cavity having a first mechanical constraint,
a second sub-cavity having a second mechanical constraint,
a third, elongated sub-cavity, wherein the first sub-cavity and the second sub-cavity are located at each end of the third, elongated sub-cavity, and
a dog-bone and/or cruciform shape;

a base including a base height that is less than the block height, wherein the base is made from a bio-inert material; and
an impermeable, removable sheet inserted into a cutout in the base and slid beneath the block.

2. The system of claim 1 , wherein the block is made from polytetrafluoroethylene (PTFE).

3. The system of claim 1 , wherein the base is made from poly lactic acid (PLA).

4. The system of claim 1 , wherein the impermeable, removable sheet is made from polytetrafluoroethylene (PTFE).

5. The system of claim 1 , wherein the impermeable, removable sheet is thinner than the base.

6. The system of claim 1 , wherein the impermeable, removable sheet is coated with silicone grease prior to insertion of the impermeable, removable sheet between the block and the base.

7. The system of claim 1 , the block width is thirty-three millimeters, and the block length is fifty-seven millimeters.

8. The system of claim 7 , wherein the base has a base width identical to the block width, a base length identical to the block length, and the base height is one and a half millimeters.

9. The system of claim 1 , wherein the base includes a rectangular cut-out spanning an entire thickness of the base and a rectangular ridge supporting the impermeable, removable sheet.

10. The system of claim 9 , wherein the rectangular ridge has a rectangular ridge height of one millimeter and a rectangular ridge width of at least one millimeter.

11. The system of claim 1 , wherein the cavity has a sample width of seven millimeters and a gauge length of twenty-one millimeters.

12. The system of claim 1 , wherein the first and second mechanical constraints are positioned at each end of the three-dimensional cavity to constrain cell driven contraction and to allow for gripping the sample after the sample is removed from the mold.

13. The system of claim 1 , wherein the first constraint and/or the second constraint comprises a natural fiber tab.

14. The system of claim 1 , wherein the first constraint and/or the second constraint comprise a porous polyethylene bar.

15. The system of claim 1 , wherein the three-dimensional cellular scaffold comprises a collagen gel.

16. The system of claim 15 , wherein the sample comprises less than forty percent of the collagen gel and more than sixty percent of the culture media.

17. The system of claim 1 , wherein the three-dimensional cellular scaffold comprises fibrin.

18. The system of claim 1 , wherein the three-dimensional cellular scaffold comprises agarose.

19. The system of claim 1 , wherein the three-dimensional cellular scaffold comprises a hydrogel.

Assignees

• Univ Boise State

Inventors

• Everingham John
• Lujan Trevor

Classifications

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N2527/00

  Culture process characterised by the use of mechanical forces, e.g. strain, vibration · CPC title

• C12N2535/00

  Supports or coatings for cell culture characterised by topography · CPC title

• C12N2533/40

  Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers · CPC title

• C12N2539/00

  Supports and/or coatings for cell culture characterised by properties · CPC title