Cell culture and gradient migration assay methods and devices

Patent metadata

Field	Value
Publication number	US-9353343-B2
Application number	US-201314081314-A
Country	US
Kind code	B2
Filing date	Nov 15, 2013
Priority date	Jan 21, 2010
Publication date	May 31, 2016
Grant date	May 31, 2016

Abstract

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

First claim

What is claimed:

1. A microfluidic device comprising:
a first flow channel containing a first substance flowing through;
a first perfusion barrier separating the first flow channel from a cell chamber, the first substance perfusing through the first perfusion barrier into the cell chamber;
a second perfusion barrier opposite from said first perfusion barrier and separating the cell chamber from a second flow channel, the second flow channel containing a second substance flowing through, the second substance perfusing through the second perfusion barrier into the cell chamber;
a gradient formed within the cell chamber comprising the first substance perfusing through the first perfusion barrier and the second substance perfusing through the second perfusion barrier;
such that a plurality of cells within the cell chamber are exposed to the gradient of the first substance and the second substance.

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 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 devices; wherein at least one well is configured so that it introduces a chemoattractant agent as the first substance into the first 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. The microfluidic device of claim 1 , further comprising:
a second gradient formed within the cell chamber, the second gradient comprising a third substance flowing through the first flow channel and perfusing through the first perfusion barrier, and a fourth substance flowing through the second flow channel and perfusing through the second perfusion barrier.

8. 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.

9. The microfluidic device of claim 1 , wherein the first substance and is chosen from the group consisting of chemoattractants, dyes, fetal bovine serum (FBS), hormonal stimuli, and drugs.

10. The microfluidic device of claim 1 , wherein the first substance is different from the second substance.

11. The microfluidic device of claim 1 , wherein the gradient has a profile that is substantially linear in a first axis perpendicular to the first perfusion barrier and second perfusion barrier, and substantially free of variation in a second axis parallel to the first perfusion barrier and second perfusion barrier.

12. The microfluidic device of claim 1 , wherein the cells in the cell chamber migrate in a profile that is substantially linear in a first axis perpendicular to the first perfusion barrier and second perfusion barrier, and migrate in a profile that is substantially free of variation in a second axis parallel to the first perfusion barrier and second perfusion barrier.

Assignees

• Emd Millipore Corp

Inventors

• Hung Ju-Sung Paul
• Lee Philip J
• Cappione Amedeo J

Classifications

• B01L2200/0647

  Handling flowable solids, e.g. microscopic beads, cells, particles · CPC title

• B01L3/502753

  characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation · CPC title

• C12M23/34

  Internal compartments or partitions · CPC title

• C12M27/18 Primary

  Flow directing inserts · CPC title

• C12M23/40

  Manifolds; Distribution pieces (fluid transfer means B01L3/563) · CPC title