Cell culture and gradient migration assay methods and devices

摘要

A number of novel improved microfluidic configurations and systems and methods of manufacture and operation for a microfluidic invasion assay system.

主权项

1. A method for culturing and monitoring cells using a microfluidic cell culture system, wherein the microfluidic cell culture system comprises a cell chamber, separated from a first microfluidic flow channel by a perfusion barrier and from a second microfluidic flow channel by a second perfusion barrier on an opposite side of the cell chamber, a gravity flow inlet well, and an outlet well, the method comprising:
introducing one or more groups of cells into the cell chamber of the microfluidic cell culture system; culturing the one or more groups of cells in the cell chamber; continuously perfusing the cultured one or more groups of cells by supplying a media solution to the gravity flow inlet well, wherein a medium level difference between the gravity flow inlet well and outlet well creates a perfusion flow rate; creating and controlling a first gradient in the cell chamber by supplying a first substance to the first microfluidic flow channel and a second substance to the second microfluidic flow channel, the first gradient forming within the cell chamber perpendicular to the first perfusion barrier and the second perfusion barrier; creating and controlling a second gradient in the cell chamber, after a selected period of time, by supplying a third substance to the first microfluidic flow channel and a fourth substance to the second microfluidic flow channel, the second gradient forming within the cell chamber perpendicular to the first perfusion barrier and the second perfusion barrier; wherein the cell chamber provides an uninterrupted optical path to the cultured one or more cells during culturing and creation and control of the gradients; capturing a plurality of images of the cultured one or more cells in the microfluidic culture system during culturing and during control of the gradients; using an information system to analyze the cultured one or more cells in a sequence of the plurality of images to determine migration properties of the cultured one or more cells in response to the first and second gradients.