Three-Dimensional Microfabricated Bioreactors with Embedded Capillary Network
First Claim
1. A method of making a microvascularized bioreactor, said method comprising:
- a. providing a photocurable liquid composition having a top surface;
b. providing a source of light capable of curing at least a portion of said composition;
c. illuminating said composition top surface with said light, wherein said illumination is in a pattern thereby simultaneously generating a polymerized pattern layer having a layer thickness;
d. immersing said polymerized pattern layer into said composition depth by a vertical displacement corresponding to said layer thickness;
e. waiting a surface dwell time for said surface to become substantially level;
f. repeating said illuminating step to generate an adjacent polymerized pattern layer, wherein said repeating step is repeated for a number of steps to generate a microvascular network having an interior surface and an exterior surface; and
g. contacting said exterior surface with a cell population, thereby obtaining said microvascularized bioreactor;
wherein said microvascular network has a permeability to a biological material that varies with location within said network.
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Accused Products
Abstract
In an aspect, the present invention uses projection micro stereolithography to generate three-dimensional microvessel networks that are capable of supporting and fostering growth of a cell population. For example, provided is a method of making a microvascularized bioreactor via layer-by-layer polymerization of a photocurable liquid composition with repeated patterns of illumination, wherein each layer corresponds to a layer of the desired microvessel network. The plurality of layers are assembled to make a microvascular network. Support structures having different etch rates than the structures that make up the network provides access to manufacturing arbitrary geometries that cannot be made by conventional methods. A cell population is introduced to the external wall of the network to obtain a microvascularized bioreactor. Provided are various methods and related bioreactors, wherein the network wall has a permeability to a biological material that varies within and along the network.
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Citations
69 Claims
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1. A method of making a microvascularized bioreactor, said method comprising:
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a. providing a photocurable liquid composition having a top surface; b. providing a source of light capable of curing at least a portion of said composition; c. illuminating said composition top surface with said light, wherein said illumination is in a pattern thereby simultaneously generating a polymerized pattern layer having a layer thickness; d. immersing said polymerized pattern layer into said composition depth by a vertical displacement corresponding to said layer thickness; e. waiting a surface dwell time for said surface to become substantially level; f. repeating said illuminating step to generate an adjacent polymerized pattern layer, wherein said repeating step is repeated for a number of steps to generate a microvascular network having an interior surface and an exterior surface; and g. contacting said exterior surface with a cell population, thereby obtaining said microvascularized bioreactor; wherein said microvascular network has a permeability to a biological material that varies with location within said network. - View Dependent Claims (2, 4, 8, 10, 11, 17, 18, 20, 25, 36, 37, 39, 68)
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3. (canceled)
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5-7. -7. (canceled)
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9. (canceled)
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12-16. -16. (canceled)
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19. (canceled)
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21-24. -24. (canceled)
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26-35. -35. (canceled)
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38. (canceled)
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40-42. -42. (canceled)
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43. A method of making a three-dimensional device, said method comprising:
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a. providing photocurable liquid composition having a top surface; b. providing a light source capable of curing at least a portion of said composition; c. illuminating said composition top surface with gray scale illumination, wherein said gray scale illumination is a pattern of light intensity or duration that generates a pattern of polymer having a spatially varying cross-linking density; and d. contacting said polymer with an etchant that selectively removes polymer having a lower cross-linking density; thereby making a three-dimensional device. - View Dependent Claims (44, 45, 47)
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46. (canceled)
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48. (canceled)
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49. A method of producing a biological material, said method comprising:
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a. providing a vascularized bioreactor having a three-dimensional network of microvessels capable of fostering a cell population, wherein said network has i. a polymeric wall with a lumen-facing side and a cell-facing side, wherein at least a portion of said wall is permeable to said biological material and said wall has a permeability that varies with microvessel position; ii. an inlet port upstream of said network for introducing culture media to said network; iii. an outlet port downstream of said network for removing said culture media; b. obtaining an isolated cell population capable of producing said biological material or a precursor thereof; c. contacting said cell population to at least a portion of said outward-facing side of said network wall; d. culturing said cell population in said bioreactor by introducing a culture media to said inlet port to expose said network wall inward-facing side to said culture media; and e. collecting said culture media that has transited said microvessel network at said outlet port; wherein said cell population produces a biological material capable of diffusing from said cell population to said culture media via said network wall so that said collecting step collects at least a portion of said produced biological material, said biological material is a biofuel, pharmaceutical, drug, a prodrug, or any precursors thereof.
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50-56. -56. (canceled)
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57. A vascularized bioreactor comprising:
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a. a three-dimensional network of microvessels having a wall made of a biocompatible polymer, wherein said wall has at least one parameter that varies with a longitudinal or a radial position within the network; b. an inlet port for introducing culture media to said network; and c. an exit port where culture media is removed from said network.
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58-67. -67. (canceled)
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69-71. -71. (canceled)
Specification