Vascularized In Vitro Perfusion Devices, Methods of Fabricating, and Applications Thereof
First Claim
1. A method of manufacturing a stable and adaptable in vitro microcirculation system, the method comprising:
- a) casting a network of channels on a polymerized matrix gel with a sacrificial material, said network cast to form at least one perfusion inlet port and inlet channel, and at least one perfusion outlet port and outlet channel, and one or more cross channels, each cross channel being in communication with both an inlet channel and an outlet channel;
b) incorporating an isolate of intact native microvessels into a polymerizable matrix;
c) distributing the polymerizable matrix from step b) over the network of cast channels and polymerizing the matrix to form a continuous polymerized gel matrix comprising both the network of cast channels and the intact native microvessels;
d) incubating the gel matrix under conditions suitable to promote spontaneous growth of a neovasculature from the native microvessel isolate, said incubating optionally taking place before or after step c);
e) flushing the sacrificial material from the cast network to yield a molded network of channels;
f) lining the molded channels with endothelial cells to form a continuous network of endothelial cell-lined channels; and
g) subjecting the network of endothelial cell-lined channels to perfusion with a perfusion fluid sufficient to induce endothelial sprouting from the one or more cross channels and inosculation between at least two sprouts and the neovasculature, thereby forming a stable adaptable microcirculation system.
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Abstract
A bona fide adaptable in vitro microcirculation model is provided by integrating a 3-D printed network of endothelial-cell lined perfusion channels, formed via sacrificial casting in a gel matrix, with a native, adaptable microvasculature matured from native microvessels added to the gel matrix. Responsive vascular adaptation exhibited by the in vitro microcirculation is physiologically relevant. Methods for fabricating, devices, models and investigative platforms for pharmaceutical applications, vascular mechanism and microvessel-parenchyma interaction studies, and vascularizing strategies for tissue engineering applications are also disclosed.
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Citations
11 Claims
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1. A method of manufacturing a stable and adaptable in vitro microcirculation system, the method comprising:
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a) casting a network of channels on a polymerized matrix gel with a sacrificial material, said network cast to form at least one perfusion inlet port and inlet channel, and at least one perfusion outlet port and outlet channel, and one or more cross channels, each cross channel being in communication with both an inlet channel and an outlet channel; b) incorporating an isolate of intact native microvessels into a polymerizable matrix; c) distributing the polymerizable matrix from step b) over the network of cast channels and polymerizing the matrix to form a continuous polymerized gel matrix comprising both the network of cast channels and the intact native microvessels; d) incubating the gel matrix under conditions suitable to promote spontaneous growth of a neovasculature from the native microvessel isolate, said incubating optionally taking place before or after step c); e) flushing the sacrificial material from the cast network to yield a molded network of channels; f) lining the molded channels with endothelial cells to form a continuous network of endothelial cell-lined channels; and g) subjecting the network of endothelial cell-lined channels to perfusion with a perfusion fluid sufficient to induce endothelial sprouting from the one or more cross channels and inosculation between at least two sprouts and the neovasculature, thereby forming a stable adaptable microcirculation system. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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Specification