Microfluidic cell culture
Inventors
Lowe, JR., Randall D. • Beaumont, Kristin G. • Karunakaran, Aathavan • Marks, Natalie C. • McEwen, Jason M. • White, Mark P. • Nevill, J. Tanner • Wang, Gang F. • McFarland, Andrew W. • Malleo, Daniele • Breinlinger, Keith J. • Guan, Xiao • Chapman, Kevin T.
Assignees
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Publication Number
US-12134758-B2
Publication Date
2024-11-05
Expiration Date
Abstract
Systems, methods and kits are described for culturing one or more biological cells in a microfluidic device, including provision of nutrients and gaseous components configured to enhance cell growth, viability, portability, or any combination thereof. In some embodiments, culturing a single cell may produce a clonal population in the microfluidic device.
Core Innovation
The invention relates to preparing and using microfluidic devices for cell growth by forming conditioned surface monolayers inside discrete microfluidic circuits. A microfluidic device is formed with a base, a microfluidic circuit structure, and a cover, each having an inner surface facing an interior of one or more microfluidic circuit elements. The inner surfaces have a monolayer covalently attached to them to support cell growth, viability, portability, or any combination thereof.
Conditioning is achieved by introducing a conditioning modification reagent comprising a moiety for supporting cell growth, viability, portability, or any combination thereof and a reactive pairing moiety Rpx into the microfluidic circuit. The monolayer includes a linking group and a reactive moiety Rx configured to covalently link to the conditioning modification reagent within the microfluidic circuit element. Covalently linking the reactive pairing moiety Rpx with the reactive moiety Rx forms a conditioned surface monolayer on the inner surfaces of the microfluidic circuit elements.
The invention further includes culturing at least one biological cell in the microfluidic device by forming the conditioned surface in a growth chamber. Cells are introduced into the growth chamber having the conditioned surface and are incubated for a period of time long enough to expand the biological cells to produce a colony, while a first fluidic medium is non-continuously perfused through a flow region during incubation.
Claims Coverage
Two independent claims are present. Across them, there are two core inventive feature sets: covalently attaching a conditioned surface monolayer inside microfluidic circuit elements using complementary reactive moieties, and culturing biological cells in a growth chamber that has the conditioned surface while applying non-continuous perfusion of a first fluidic medium.
Covalently forming a conditioned surface monolayer via complementary reactive moieties
Introducing a conditioning modification reagent having a moiety for supporting cell growth, viability, portability, or any combination thereof and a reactive pairing moiety Rpx into a microfluidic circuit, wherein a monolayer on the inner surfaces includes a linking group and a reactive moiety Rx configured to covalently link to the conditioning modification reagent, and wherein covalently linking the reactive pairing moiety Rpx with the reactive moiety Rx forms a conditioned surface monolayer on the inner surfaces of the microfluidic circuit elements so the conditioned surface monolayer is configured to support cell growth, viability, portability, or any combination thereof.
Conditioned-surface culturing using a discrete microfluidic circuit and non-continuous perfusion
Forming a conditioned surface in a growth chamber by introducing a conditioning modification reagent, wherein inner surfaces have a covalently attached monolayer comprising a linking group and a reactive moiety Rx that covalently links to the conditioning modification reagent, introducing at least one biological cell into the growth chamber configured to support cell growth, viability, portability, or any combination thereof, incubating for a period of time long enough to expand the biological cell to produce a colony, and non-continuously perfusing the first fluidic medium through a flow region during incubation.
Overall, the claim coverage focuses on preparing microfluidic devices with covalently attached conditioned surface monolayers formed by complementary reactive moieties, and on culturing biological cells in a growth chamber within a discrete microfluidic circuit using non-continuous perfusion during incubation.
Stated Advantages
Configured to support cell growth, viability, portability, or any combination thereof within the microfluidic device.
Enables expansion of biological cells to produce a colony during incubation.
Documented Applications
Preparing a microfluidic device to support cell growth, viability, portability, or any combination thereof by forming a conditioned surface monolayer in microfluidic circuit elements.
Culturing at least one biological cell in a microfluidic device, incubating to expand cells to produce a colony of biological cells using the conditioned surface in a growth chamber and non-continuous perfusion through a flow region.
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