FABRICATION OF TISSUE LAMINA USING MICROFABRICATED TWO-DIMENSIONAL MOLDS

• US 20100098742A1
• Filed: 06/28/2002
• Published: 04/22/2010
• Est. Priority Date: 04/30/1999
• Status: Active Grant

• Current Assignee: Charles Stark Draper Laboratory Incorporated, The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: Charles Stark Draper Laboratory Incorporated
• Inventors: Sevy, Alexander, Maemura, Tomoyuki, Wang, Chiao-Chun, Borenstein, Jeffrey T., Kaazempur-Mofrad, Mohammad Reza, Vacanti, Joseph P., Weinberg, Eli, Ishii, Osamu, Ogilvie, Jennifer, King, Kevin R., Shin, Young-Moon Michael
• Granted Patent: US 7,759,113 B2
• Time in Patent Office: Days
• US Class Current: 424/424
• 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

1-117. -117. (canceled)

118. A tissue lamina having multiple layers, each layer comprising a first polymer scaffold configured with a plurality of branched microchannels, a second polymer scaffold having a reservoir holding animal cells, and a semi-permeable membrane disposed between the first and second polymer scaffolds, the semi-permeable membrane configured for allowing exchange of oxygen, nutrients and waste between circulating fluid flowing in the plurality of microchannels and animal cells held in the reservoir, wherein the first polymer scaffold and the second polymer scaffold are fastened together about the semi-permeable membrane such that the circulating fluid indirectly passes oxygen, nutrients and waste with the animal cells, thereby making a tissue lamina that can supplement or replace at least one organ function.

124. The tissue lamina of claim 118, wherein at least a portion of each of the first and second polymer scaffolds have microchannels for the circulating fluid that form a pattern, and the pattern in the first polymer scaffold is identical to the pattern in the second polymer scaffold and designed to efficiently aid the exchange of oxygen, nutrients and waste.

125. The tissue lamina of claim 118, wherein at least a portion of each of the first and second polymer scaffolds have microchannels for the circulating fluid that form a pattern, and the pattern in the first polymer scaffold is non-identical to the pattern in the second polymer scaffold and designed to efficiently aid the exchange of oxygen, nutrients and waste.

126. The tissue lamina of claim 118, wherein the microchannels have a minimum diameter of from about 1 μ

m to about 500 μ

m.

127. The tissue lamina of claim 118, wherein at least one growth factor is covalently linked to at least one of the polymer scaffolds.

128. The tissue lamina of claim 118, wherein at least one growth factor is covalently linked to the semi-permeable membrane.

129. The tissue lamina of claim 118, wherein the semi-permeable membrane is biodegradable.

130. The tissue lamina of claim 118, wherein the semi-permeable membrane has pores of between about 0.01 and about 20 μ

m.

131. The tissue lamina of claim 118, wherein at least one of the polymer scaffolds comprises through-holes for receiving fasteners to secure the tissue lamina together.

132. The tissue lamina of claim 118, wherein the first and second polymer scaffolds are fastened by solvent bonding, reflow by heating, treating the surface of the polymer scaffolds with oxygen plasma, polymer flow at the surface of the polymer scaffolds, mechanically fastening the polymer scaffolds with fasteners, or adhering the polymer scaffolds with adhesives, or adhesive films.

134. The tissue lamina of claim 118, wherein the animal cells are selected from the group consisting of endothelial cells, parenchymal cells, bone marrow cells, hematopoietic cells, muscle cells, osteoblasts, stem cells, and fibroblasts.

135. The tissue lamina of claim 118, further comprising endothelial cells seeded in the microchannels of the first polymer scaffold and the animal cells of the second polymer scaffold comprises parenchymal cells.

136. The tissue lamina of claim 135, wherein the parenchymal cells are derived from stem cells or from an organ selected from the group consisting of heart, liver, pancreas, intestine, brain, kidney, reproductive tissue, lung, muscle and bone marrow.

137. The tissue lamina of claim 136, wherein the organ is liver.

138. The tissue lamina of claim 136, wherein the organ is kidney.

139. The tissue lamina of claim 118, wherein the material of the first polymer scaffold is identical to the material of the second polymer scaffold.

141. The tissue lamina of claim 118, wherein the material of at least one polymer scaffold is biodegradeable.

235. The tissue lamina of claim 135, wherein the parenchymal cells are derived from stem cells.

243. The tissue lamina of claim 118, wherein the first polymer scaffold is comprised of a material selected from the group consisting of poly(dimethyl siloxane), poly(glycerol-sebacate), polylactide-co-glycolide, and combinations thereof.

244. The tissue lamina of claim 118, wherein the second polymer scaffold is comprised of a material selected from the group consisting of poly(dimethyl siloxane), poly(glycerol-sebacate), polylactide-co-glycolide, and combinations thereof.

272. The tissue lamina apparatus of claim 118, wherein the semi-permeable membrane is comprised of a material that is suitable for attachment and culturing of animal cells.

276. The tissue lamina of claim 118, wherein the animal cells are human cells.

288. The tissue lamina of claim 118, wherein the first polymer scaffold comprises a molded polymer scaffold.

289. The tissue lamina of claim 118, wherein the second polymer scaffold comprises a molded polymer scaffold.

290. The tissue lamina of claim 118, wherein the tissue lamina is implantable.

291. The tissue lamina of claim 118, wherein one or more of the plurality of branched microchannels are open-faced channels, wherein open portions of the channels are adjacent to the semi-permeable membrane.

292. The tissue lamina of claim 118, wherein the reservoir is an open-faced compartment adjacent to the semi-permeable membrane.

293. The tissue lamina of claim 118, wherein the material of the first polymer scaffold is non-identical to the material of the second polymer scaffold.

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168. A method of making a tissue lamina comprising the steps of:

(a) positioning a semi-permeable membrane between a first and second polymer scaffold, wherein the first polymer scaffold is configured with a plurality of branched microchannels and the second polymer scaffold is configured as a reservoir for holding animal cells;

(b) fastening the first and second polymer scaffolds together such that the semi-permeable membrane is disposed between the polymer scaffolds;

(c) seeding animal cells into the reservoir of the second polymer scaffold;

(d) indirectly exchanging oxygen, nutrients and waste between a fluid flowing in the plurality of microchannels and animal cells held in the reservoir through the semi-permeable membrane; and

(e) culturing the animal cells under conditions such that tissue is formed to supplement or replace at least one organ function.

173. The method of claim 168, wherein at least a portion of each of the first and second polymer scaffolds have microchannels for the circulating fluid that form a pattern, and the pattern in the first polymer scaffold is identical to the pattern in the second polymer scaffold and designed to efficiently aid the exchange of oxygen, nutrients and waste.

174. The method of claim 168, wherein at least a portion of each of the first and second polymer scaffolds have microchannels for the circulating fluid that form a pattern, and the pattern in the first polymer scaffold is non-identical to the pattern in the second polymer scaffold and designed to efficiently aid the exchange of oxygen, nutrients and waste.

175. The method of claim 168, wherein the microchannels have a minimum diameter of from about 1 μ

m to about 500 μ

m.

176. The method of claim 168, wherein at least one growth factor is covalently linked to at least one of the polymer scaffolds.

177. The method of claim 168, wherein at least one growth factor is covalently linked to the semi-permeable membrane.

178. The method of claim 168, wherein the semi-permeable membrane is biodegradable.

179. The method of claim 168, wherein the semi-permeable membrane has pores of between about 0.01 and about 20 μ

m.

180. The method of claim 168, wherein at least one of the polymer scaffolds comprises through-holes for receiving fasteners to secure the tissue lamina together.

181. The method of claim 168, wherein the first and second polymer scaffolds are fastened by solvent bonding, reflow by heating, treating the surface of the polymer scaffolds with oxygen plasma, polymer flow at the surface of the polymer scaffolds, mechanically fastening the polymer scaffolds with fasteners, or adhering the polymer scaffolds with adhesives, or adhesive films or adhesive molds.

183. The method of claim 168, wherein the animal cells are selected from the group consisting of endothelial cells, parenchymal cells, bone marrow cells, hematopoietic cells, muscle cells, osteoblasts, stem cells, and fibroblasts.

184. The apparatus of claim 168, wherein the first polymer scaffold comprises endothelial cells seeded in the microchannels of the first polymer scaffold and the animal cells of the second polymer scaffold are parenchymal cells.

185. The method of claim 184, wherein the parenchymal cells are derived from an organ selected from the group consisting of heart, liver, pancreas, intestine, brain, kidney, reproductive tissue, lung, muscle and bone marrow.

186. The method of claim 185, wherein the organ is liver.

187. The method of claim 185, wherein the organ is kidney.

188. The method of claim 168, wherein the material of the first polymer scaffold is identical to the material of the second polymer scaffold.

189. The method of claim 168, wherein the material of the first polymer scaffold is non-identical to the material of the second polymer scaffold.

190. The method of claim 168, wherein the material of at least one polymer scaffold is biodegradeable.

192. The method of claim 168, wherein the animal cells are parenchymal cells.

237. The method of claim 184, wherein the parenchymal cells are derived from stem cells.

248. The method of claim 168, wherein the second polymer scaffold is comprised of a material selected from the group consisting of poly(dimethyl siloxane), poly(glycerol-sebacate), polylactide-co-glycolide, and combinations thereof.

274. The method of claim 168, wherein the semi-permeable membrane is comprised of a material that is suitable for attachment and culturing of animal cells.

278. The method of claim 168, wherein the animal cells are human cells.

282. The method of claim 168, wherein the first polymer scaffold comprises circulatory flow.

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