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US 2013/0171682 A1Published application
Cell Culture and Gradient Migration Assay Methods and Devices
AssigneeEMD Millipore Corporation
InventorsJu-Sung Paul Hung, Philip J. Lee, Amedeo J. Cappione, III
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Abstract
A number of novel improved microfluidic configurations and systems and methods of manufacture and operation for a microfluidic invasion assay system.
Claims33
A method for culturing and monitoring cells using a microfluidic cell culture system comprising:
- introducing one or more groups of cells in to a cell chamber of the microfluidic cell culture system;
- culturing the one or more groups of cells in the cell chamber;
- creating and controlling a gradient in the cell chamber using one or more microfluidic flow channels;
- wherein the cell chamber provides an uninterrupted optical path to the cells during culturing and creation and control of the gradient;
- capturing a plurality of images of the cells in the microfluidic culture system during culturing and during control of the gradient;
- using an information system to analyze one or more cells in a sequence of the plurality of images to determine migration properties of the one or more cells.
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A method for culturing and monitoring cells using microfluidic cell culture systems comprising:
- monitoring cells (e.g., the metastatic human breast cancer cell line MDA-MB-231) to determine their movement in response to spatial changes in serum concentration;
- detecting for cells exposed to a gradient (e.g. FBS), movement of cells along the Y-axis toward the upper barrier of the cell chamber in the positive direction of the gradient;
- monitoring one or more control cells cultured in the absence of a spatial serum gradient to detect less movement with far more random directional properties;
- wherein cells exposed to gradients are determined to move preferentially toward the source of higher gradient concentration;
- and wherein both cell expansion and movement occurred during the 4 hour to 6 day intervals, with images taken some period after gradient induction.
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A microfluidic device comprising:
- a. a first channel;
- b. a first barrier separating the first channel from a cell chamber;
- c. a second barrier separating the cell chamber from a second channel;
- d. the first channel configured to contain a first substance;
- d. the second channel configured to contain a second substance;
- e. such that cells in the cell holding region are exposed to a gradient of the first substance in the cell chamber attracting the cells towards the first barrier.
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Dependencies are inferred from the claim text.