Fabrication of vascularized tissue using microfabricated two-dimensional molds

• US 10,231,820 B2
• Filed: 11/17/2016
• Issued: 03/19/2019
• Est. Priority Date: 03/25/2002
• Status: Expired due to Fees

• Current Assignee: Charles Stark Draper Laboratory Incorporated
• Original Assignee: Charles Stark Draper Laboratory Incorporated, The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Vacanti, Joseph P., Shin, Young-Moon M., Ogilvie, Jennifer, Sevy, Alexander, Maemura, Tomoyuki, Ishii, Osamu, Kaazempur-Mofrad, Mohammad R., Borenstein, Jeffrey T., King, Kevin R., Wang, Chiao-Chun, Weinberg, Eli
• Primary Examiner(s): Fox, Allison M
• Application Number:
  US15/354,865
• Publication Number:
  US 20170296322A1
• Time in Patent Office: 852 Days
• Field of Search: None
• CPC Class Codes: A61F 2/062 Apparatus for the production of blood vessels made from natural tissue or with layers of living cells prostheses made from natural tissue or living cells A61L27/507
• CPC Class Codes: A61F 2240/004 Using a positive or negative model, e.g. moulds
• CPC Class Codes: A61F 2240/005 Templates
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3808 Endothelial cells
• CPC Class Codes: A61L 27/3891 as distinct cell layers
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/06 Tubular C12M23/08, C12M23/16 take precedence
• CPC Class Codes: C12M 23/30 biodegradable
• CPC Class Codes: C12M 25/02 Membranes; Filters filters or filtration in general B01D24/00-B01D41/00
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture
• CPC Class Codes: C12N 2533/12 Glass
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691
• CPC Class Codes: Y10T 29/49826 : Assembling or joining

Abstract

Methods and materials for making complex, living, vascularized tissues for organ and tissue replacement, especially complex and/or thick, structures, such as liver tissue is provided. Tissue lamina is made in a system comprising an apparatus having (a) a first mold or polymer scaffold, a semi-permeable membrane, and a second mold or polymer scaffold, wherein the semi-permeable membrane is disposed between the first and second molds or polymer scaffolds, wherein the first and second molds or polymer scaffolds have means defining microchannels positioned toward the semi-permeable membrane, wherein the first and second molds or polymer scaffolds are fastened together; and (b) animal cells. Methods for producing complex, three-dimensional tissues or organs from tissue lamina are also provided.

Claims

1. A lamina assembly comprising:

a lower structure having an upper side defining lower structure channels;

an upper structure having a lower side defining at least one upper structure microchannel that opposes the lower structure channels;

a semi-permeable membrane disposed between the lower and upper structures to separate the respective channels; and

cells cultured on at least one side of the semi-permeable membrane.

2. A lamina assembly as recited in claim 1, wherein the lower and upper structures are fastened together.

3. A lamina assembly as recited in claim 1, wherein each of the first and second structures and the semi-permeable membrane are comprised of material that is suitable for attachment and culturing of animal cells and the cells are animal cells.

4. A lamina assembly as recited in claim 1, wherein the cells are cultured on both sides of the semi-permeable membrane.

5. A lamina assembly as recited in claim 1, wherein the lower structure channels form an inlet in fluid communication with an outlet through a plurality of branching channels.

6. A lamina assembly as recited in claim 5, further comprising a nutrient supply connected to the inlet and an excretion removal line connected to the outlet.

7. A lamina assembly as recited in claim 5, further comprising a pump connected to the inlet for circulating fluid through the tissue lamina assembly.

8. A lamina assembly as recited in claim 5, wherein the inlet is directly connected to blood vessels of an animal.

9. A lamina assembly as recited in claim 1, wherein in the upper structure at least one microchannel forms a branching pattern identical to a branching pattern in the lower structure.

10. A lamina assembly as recited in claim 1, wherein the upper structure has an upper side defining upper structure channels, and

further comprising:

a second upper structure having a lower side defining second upper structure channels that oppose the upper structure channels of the upper side; and

a second semi-permeable membrane disposed between the upper and second upper structures to separate the respective channels; and

cells cultured on at least one side of the second semi-permeable membrane.

11. A lamina assembly as recited in claim 10, wherein the second upper structure has an upper side defining second upper structure channels, and

further comprising:

a third upper structure having a lower side defining third upper structure channels that oppose the second upper structure channels of the upper side; and

a third semi-permeable membrane disposed between the second and third upper structures to separate the respective channels; and

cells cultured on at least one side of the third semi-permeable membrane.

12. A lamina assembly as recited in claim 1, wherein the lower structure channels are open-faced channels and open portions of the lower structure channels are adjacent to the semi-permeable membrane.

13. A lamina assembly as recited in claim 1, further comprising a second tissue lamina assembly attached to the upper structure, the second tissue lamina assembly including:

a third structure having a side defining third structure channels;

a fourth structure having a side defining fourth structure channels that oppose the third structure channels;

a second semi-permeable membrane disposed between the third and fourth structures to separate the respective channels; and

cells cultured on at least one side of the semi-permeable membrane.

14. A lamina assembly for supplementing or replacing at least one organ function, comprising:

a lower structure having an upper side defining a plurality of lower microchannels;

an upper structure having a lower side defining at least one channel that opposes at least a portion of the lower microchannels;

a semi-permeable membrane disposed between the lower microchannels and the at least one channel; and

cells cultured on at least one of the semi-permeable membrane, the lower structure or the upper structure.

15. A lamina assembly as recited in claim 14, wherein the plurality of lower microchannels have endothelial cells seeded therein and the at least one channel has parenchymal cells therein.

16. A lamina assembly as recited in claim 14, wherein the semi-permeable membrane allows exchange of oxygen, nutrients and waste between fluid circulating in the plurality of lower microchannels and the at least one channel.

17. A lamina assembly as recited in claim 14, wherein each of the lower and upper structures and the semi-permeable membrane are comprised of material that is suitable for attachment and culturing of animals cells, and wherein animal cells are cultured on both sides of the semi-permeable membrane.

18. A lamina assembly as recited in claim 14, wherein the lower and upper structures are fastened together and the plurality of lower microchannels form at least one inlet in communication with at least one outlet.

19. A lamina assembly as recited in claim 14, further comprising cells cultured on at least one side of the semi-permeable membrane, wherein the plurality of lower microchannels are branched microchannels.