Microfluidic device for generating neural cells to simulate post-stroke conditions

Patent metadata

Field	Value
Publication number	US-9632076-B2
Application number	US-201414539544-A
Country	US
Kind code	B2
Filing date	Nov 12, 2014
Priority date	Feb 3, 2012
Publication date	Apr 25, 2017
Grant date	Apr 25, 2017

Abstract

This application provides devices for modeling ischemic stroke conditions. The devices can be used to culture neurons and to subject a first population of the neurons to low-oxygen conditions and a second population of neurons to normoxic conditions. The neurons are cultured on a porous barrier, and on the other side of the barrier run one or more fluid-filled channels. By flowing fluid with different oxygen levels through the channels, one can deliver desired oxygen concentrations to the cells nearest those channels.

First claim

What is claimed:

1. A method of modeling an ischemic condition, comprising:
providing a microfluidic device including a fluid-filled chamber, a channel, a porous barrier separating the chamber from the channel, a first population of CNS cells proximal to the channel, and a second population of CNS cells distal from the channel relative to the first population of CNS cells;
concurrently with exposing the first population of CNS cells to a low concentration of oxygen, exposing the second population of CNS cells to a high concentration of oxygen, thereby modeling the ischemic condition, wherein:
exposing the first population of CNS cells to a low concentration of oxygen comprises flowing an oxygen scavenger through the channel; and
measuring at least one cellular property of the first population of CNS cells.

2. The method of claim 1 , wherein ischemic condition is ischemic stroke.

3. The method of claim 1 , wherein the CNS cells comprise neurons, microglia, astrocytes, oligodendrocytes, or neural progenitors.

4. The method of claim 1 , further comprising producing a third population of CNS cells between the first population and the second population, wherein the third population of CNS cells models a penumbra produced by an ischemic stroke.

5. The method of claim 1 , further comprising measuring trans-endothelial electrical resistance across the first and second population of cells.

6. The method of claim 1 , further comprising visualizing the cells in the device by microscopy.

7. The method of claim 1 , further comprising removing the cells from the device and performing biochemical analysis or microscopy on the removed cells.

8. The method of claim 1 , wherein measuring the at least one cellular property further comprises testing for a factor secreted by the cells.

9. The method of claim 8 , comprising measuring for the factor in a sample taken from the chamber.

10. The method of claim 8 , comprising measuring for the factor in a sample taken from the channel.

11. The method of claim 1 , further comprising exposing the cells to a test agent.

12. The method of claim 11 , wherein the test agent promotes neurodegeneration, necrosis, or apoptosis.

13. The method of claim 11 , wherein the agent is a cancer cell capable of invading neural tissue.

14. The method of claim 1 , further comprising not perfusing a fluid through the fluid-filled chamber.

15. A method of modeling an ischemic condition, comprising:
providing a microfluidic device including a fluid-filled chamber, a channel, a porous barrier separating the chamber from the channel, a first population of CNS cells proximal to the channel, and a second population of CNS cells distal from the channel relative to the first population of CNS cells;
concurrently with exposing the first population of CNS cells to a high concentration of oxygen, exposing the second population of CNS cells to a low concentration of oxygen, thereby modeling the ischemic condition, wherein:
exposing the first population of CNS cells to a high concentration of oxygen comprises flowing oxygenated fluid through the channel; and

measuring at least one cellular property of the first population of CNS cells.

Assignees

• Charles Stark Draper Laboratory Inc
• The Univ Of South Florida

Inventors

• Achyuta Anilkumar Harapanahalli
• Cuevas Javier
• Sundaram Shivshankar
• Katnik Chris

Classifications

• G01N33/5058 Primary

  Neurological cells · CPC title

• G01N2510/00

  Detection of programmed cell death, i.e. apoptosis · CPC title

• G01N33/5026

  on cell morphology · CPC title

• C12M41/34

  of gas · CPC title

• G01N2333/545

  IL-1 · CPC title