Cell culture and gradient migration assay methods and devices
A number of novel improved microfluidic configurations and systems and methods of manufacture and operation for a microfluidic invasion assay system.
1. A microfluidic device comprising:
a first flow channel, the first flow channel configured so that a first substance flows therein;
a first perfusion barrier separating the first flow channel from a cell chamber;
a second flow channel, the second flow channel being configured so that a second substance flows therein;
a second perfusion barrier separating the cell chamber from the second flow channel;
such that cells in the cell chamber are exposed to a gradient of the first substance in the cell chamber attracting the cells towards the first perfusion barrier.
2. A microfluidic device according to claim 1 , wherein the first flow channel, second flow channel, or both flow channels are separated from the cell chamber by a set of cross section microfluidic pores or passages measuring about 8×8 microns.
3. A microfluidic device according to claim 1 , wherein the first perfusion barrier and second perfusion barrier consist of a network of passages of approximately 100×4×2 μm (L×W×H) and further wherein a narrow cross section of the passages prevents cells from passing through the first perfusion barrier and the second perfusion barrier.
4. A microfluidic device according to claim 1 , wherein the cell chamber is about 4.8×0.5×0.05 mm in dimension (L×W×H) and is used to count the number of cells that migrate.
5. A microfluidic system comprising:
one or more devices according to claim 1 ;
an active control microfluidic plate with at least three inlet wells connected to each of the one or more devices; wherein at least one well is configured so that it introduces a chemoattractant agent as the first substance into the first flow channel; and
a manifold for controlling pressure and/or contents in at least one well connected to at least one device during cell culture.
6. The system according to claim 5 , further comprising:
at least three inlet wells and one outlet well associated with each of the one or more devices.
7. An active control microfluidic plate comprising:
at least one microfluidic device according to claim 1 ;
a first inlet well and a second inlet well in communication with the first flow channel of the microfluidic device;
a third inlet well and a fourth inlet well in communication with the second flow channel of the microfluidic device;
the first and second inlet wells configured to receive the first substance, and the third and fourth inlet wells configured to receive the second substance.
8. The microfluidic device of claim 1 , wherein the first substance is chosen from the group consisting of chemoattractants, dyes, fetal bovine serum (FBS), hormonal stimuli, and drugs.
9. The microfluidic device of claim 1 , wherein the first substance is different from the second substance.
10. The microfluidic device of claim 1 , wherein the gradient has a profile that is linear in a first axis perpendicular to the first perfusion barrier and second perfusion barrier, and free of variation in a second axis parallel to the first perfusion barrier and second perfusion barrier.
11. The microfluidic device of claim 1 , wherein the cells in the cell chamber migrate in a profile that is linear in a first axis perpendicular to the first perfusion barrier and second perfusion barrier, and migrate in a profile that is free of variation in a second axis parallel to the first perfusion barrier and second perfusion barrier.