Synthetic microfluidic systems for hypoxia
Inventors
Prabhakarpandian, Balabhaskar • Pant, Kapil
Assignees
Publication Number
US-10775364-B2
Publication Date
2020-09-15
Expiration Date
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Abstract
A method of inducing ischemia includes: providing a cell culture device having a first cell culture in an internal chamber and a second cell culture in at least one fluid channel and a perfusion modulating system that causes changes in oxygen flow in the internal chamber and/or at least one fluid channel; flowing liquid media having oxygen through the internal chamber and at least one fluid channel of the cell culture device; modulating oxygen perfusion in the internal chamber with the perfusion modulating system by varying and selectively blocking the flow rate of at least one of the liquid media or oxygen through the internal chamber to induce varying levels of ischemia; and assaying for ischemia in the first cell culture.
Core Innovation
A method of inducing ischemia comprises providing a cell culture device having a first cell culture in an internal chamber and a second cell culture in at least one fluid channel, the cell culture device comprising an internal chamber with at least one port coupled to a perfusion modulating system, at least one fluid channel bordering the internal chamber with at least one port coupled to the perfusion modulating system, and a wall separating the internal chamber and at least one fluid channel having gaps that fluidly couple the internal chamber with the at least one fluid channel. The method includes flowing liquid media having oxygen through the internal chamber and at least one fluid channel, modulating oxygen perfusion in the internal chamber by varying and selectively blocking the flow rate of at least one of the liquid media or oxygen through the internal chamber to induce varying levels of ischemia, and assaying for ischemia in the first cell culture.
The invention addresses limitations of existing ischemia and hypoxia models by providing an in vitro microfluidic device and assay for studying ischemia and hypoxia on cells in controlled perfusion conditions. The background describes that current animal models are expensive, technically complex and raise ethical concerns, and that existing in vitro models have limitations, making it advantageous to have devices and methods that provide improved experimental analysis and studies on cells and cultures under ischemic and hypoxic conditions.
The device is described as a microfluidic chip having a central internal chamber surrounded by capillary channels separated by posts or pillar structures with gaps to allow diffusion-based fluidic coupling, and can be configured as synthetic microvascular networks (SMN) or idealized microvascular networks (IMN). The device is described for use in simulating ischemia and hypoxia by interrupting flow of media and oxygen, enabling controlled study of ischemia and hypoxia, interaction between endothelial cells and tissue, and use in therapeutic screening and assays for various cell types including myocytes, neurons, nephrons, and hepatocytes.
Claims Coverage
This coverage identifies seven main inventive features drawn from the single independent claim (claim 1) and its central steps and device elements.
Internal chamber configured for an internal cell culture
The internal chamber being configured for an internal cell culture that has at least one port coupled to a perfusion modulating system capable of modulating perfusion of media in the internal chamber.
Fluid channel bordering the internal chamber configured for a channel cell culture
The at least one fluid channel bordering the internal chamber, the at least one fluid channel being configured for a channel cell culture that has at least one port coupled to the perfusion modulating system capable of modulating perfusion of media in the at least one fluid channel.
Wall separating internal chamber and fluid channel with gaps
A wall separating the internal chamber and at least one fluid channel having gaps that fluidly couple the internal chamber with the at least one fluid channel.
Perfusion modulating system causing changes in oxygen flow
Wherein the perfusion modulating system causes changes in oxygen flow in the internal chamber and/or at least one fluid channel.
Flowing liquid media having oxygen through chamber and channels
Flowing liquid media having oxygen through the internal chamber and at least one fluid channel of the cell culture device.
Modulating oxygen perfusion by varying and selectively blocking flow
Modulating oxygen perfusion in the internal chamber with the perfusion modulating system by varying and selectively blocking the flow rate of at least one of the liquid media or oxygen through the internal chamber to induce varying levels of ischemia.
Assaying for ischemia in the first cell culture
Assaying for ischemia in the first cell culture.
The independent claim centers on a microfluidic cell culture device with an internal chamber and bordering fluid channel(s) coupled to a perfusion modulating system, a porous separating wall, active modulation of oxygen-containing media including selective blocking to induce varying ischemia, and subsequent assaying for ischemia.
Stated Advantages
Provides a more physiologically-relevant testing system for therapeutic screening and basic research of ischemia and hypoxia.
Enables creation of small volume, inexpensive, disposable, optically clear microfluidic chips suitable for long-term cell culture and cellular assays.
Allows perfusion, occlusion, and reperfusion at flow rates observed in live capillaries and introduction of chemical or physical insults to cells.
Permits real-time visualization and is amenable to medium to high throughput screening, including integration into well plate formats.
Facilitates study of differences between healthy and diseased microvasculature and interaction between endothelial cells and tissue.
Can reduce animal studies during preliminary stages of therapeutic development and provides a valid and cost-effective tool for studying new therapeutic approaches.
Documented Applications
Studying ischemia and hypoxia on cells in controlled perfusion conditions.
Development and screening of therapeutics for treating ischemia and hypoxia affected tissues.
High-throughput assays for studying any cell having ischemia or hypoxia and integration into well plate formats.
Modeling and studying myocardial ischemia (MI) in a physiological microenvironment with myocytes in the internal chamber.
Studying ischemia and hypoxia in other tissue cell types, including neurons, nephrons, and hepatocytes.
Creating gradients of hypoxia, inducing reperfusion injury, and simulating varying intensities and durations of ischemia.
Co-culture assays with endothelial cells in capillary channels to study interactions with tissue cells.
Characterizing viability, pH/hypoxia, and biomarkers in ischemia, hypoxia, and post-MI conditions.
Studying stem cell therapies and regenerative approaches for cells in the internal chamber.
Using the device for drug discovery and mechanistic characterization of ischemia and hypoxia stimuli.
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