Models and methods to establish perfused vascularized tissues in three-dimensional in vitro culture
AssigneeAdvanced Solutions Life Sciences, LLC
InventorsJames B. Hoying, Michael W. Golway, Sarah Moss
Life of this patent
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Abstract
Provided herein are 3D tumor angiogenesis models and their methods of preparation and use. In some aspects, the need for identifying whether a potential drug target influences angiogenesis, identifying compounds that modulate angiogenesis, and identifying new drug targets for modulating angiogenesis.
Claims36
A three-dimensional (3D) tumor model comprising:
- tumor cells;
- and,
- isolated microvessel fragments or a microvasculature developed therefrom,
- wherein the isolated microvessel fragments or the microvasculature are embedded within a polymerized medium comprised of extracellular matrix.
A method for preparing a vascularized 3D tumor model comprising:
- providing isolated microvessel fragments to a space between two channels embedded within a polymerized medium;
- and
- providing tumor cells on or embedded within the polymerized medium.
A method for preparing a vascularized 3D tumor model comprising:
- providing isolated microvessel fragments to a space between a first channel and a second channel embedded within a polymerized medium;
- incubating the isolated microvessel fragments within the polymerized medium for a period until angiogenesis is observed;
- perfusing a fluid media from an inlet reservoir through the first channel to an outlet reservoir, wherein the outlet reservoir is operably connected to the second channel such that the perfused fluid media can traverse the second channel, wherein the fluid media is perfused for about three to five days or until the isolated microvessel fragments have visibly inosculated;
- at least partially obscuring the first channel at the outlet to the outlet reservoir to provide an increased pre-load pressure;
- re-initiating perfusion of the fluid media;
- and
- providing tumor cells on or embedded within the polymerized medium.
A method for preparing a vascularized 3D model comprising:
- providing isolated microvessel fragments to a space between a first channel and a second channel embedded within a polymerized medium;
- incubating the isolated microvessel fragments within the polymerized medium for a period until angiogenesis is observed;
- perfusing a fluid media from an inlet reservoir through the first channel to an outlet reservoir, wherein the outlet reservoir is operably connected to the second channel such that the perfused fluid media can traverse the second channel, wherein the fluid media is perfused for about three to five days or until the isolated microvessel fragments have visibly inosculated;
- at least partially obscuring the first channel at the outlet to the outlet reservoir to increase channel preload;
- providing isolated endothelial cells to the first and/or second channels;
- and
- re-initiating perfusion of the fluid media.
A 3D angiogenesis model comprising isolated microvessel fragments or a microvasculature developed therefrom between two parallel channels embedded within a polymerized medium, wherein each channel comprises an inlet end and an outlet end, each inlet end being operably connected to a fluid media source and wherein at least one outlet end is operably linked to the inlet end of a different channel and wherein the fluid media is actively pumped into at least one channel to allow for interstitial flow-conditioning.
Dependencies are inferred from the claim text.