Capillary perfused bioreactors with multiple chambers

• US 7,534,601 B2
• Filed: 08/27/2003
• Issued: 05/19/2009
• Est. Priority Date: 08/27/2002
• Status: Expired due to Fees

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Prokop, Alex, Wikswo, John P., Lin, Charles P., Hofmeister, William H., Leboeuf, Eugene J., Reiserer, Randall S., Chung, Chang Y., Cliffel, David, Haselton, Frederick R., Stremler, Mark A., McCawley, Lisa J., Baudenbacher, Franz J.
• Primary Examiner(s): Lam; Ann Y
• Application Number:
  US10/525,559
• Publication Number:
  US 20060141607A1
• Time in Patent Office: 2,092 Days
• Field of Search: 435/289.1, 435/4, 435/7.92, 4352871-2875, 435/7.2, 436/514, 422/249, 422/246
• US Class Current: 435/289.1
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 29/10 Perfusion

Abstract

A bioreactor for cultivating living cells in a liquid medium. In one embodiment of the present invention, the bioreactor includes a first substrate having a first surface, an opposite second surface and edges. The bioreactor further includes a second substrate having a first surface and an opposite second surface, defining a cavity with a bottom surface, where the bottom surface is located therebetween the first surface and the second surface. The first surface of the first substrate is received by the second surface of the second substrate to cover the cavity so as to form a channel for receiving cells and a liquid medium. In forming the bioreactor, the channel is sized to allow the growth of a layer of cells on a biocompatible coating layer and a flow of liquid in the channel. The flow of liquid is controlled so as to provide a known shear force to the layer of cells. The flow of liquid can be further controlled so as to provide an environment that simulates a vascular space in the channel.

Claims

1. A bioreactor comprising:

i. a first substrate having a first surface and an opposite second surface, defining a chamber therebetween the first surface and the opposite second surface of the first substrate for receiving biological cells and a liquid medium;

ii. an inlet port formed in the first substrate and apart from the chamber;

iii. a first connection channel formed in the first substrate, wherein the first connection channel is in fluid communication with the inlet port and the chamber for allowing a stream of substance to be delivered to the chamber;

iv. an outlet port formed in the first substrate and apart from the chamber;

v. a second connection channel formed in the first substrate, wherein the second connection channel is in fluid communication with the outlet port and the chamber for allowing a stream of substance to be removed from the chamber; and

vi. confining means positioned in the chamber to form a confinement region to confine the biological cells therein with the liquid medium,

wherein the chamber, the inlet portion, the first connection channel, the outlet port, and the second connection channel are all formed in the first substrate;

wherein the confining means comprises a first filter and a second filter, wherein the first filter is positioned proximate to the first connection channel and the second filter is positioned proximate to the second connection channel, and the first filter and the second filter are substantially parallel to each other; and

wherein each of the first filter and the second filter comprises an array of posts spaced apart from each other not to allow biological cells to pass through it.

2. The bioreactor of claim 1, wherein the first substrate is fabricated from glass, Mylar, PDMS, silicon, a polymer, a semiconductor, or any combination of them.

3. The bioreactor of claim 1, further comprising a second substrate having a first surface and an opposite, second surface, wherein the second substrate is sized such that when the first surface of the second substrate is received by the second surface of the first substrate, the chamber is covered.

4. The bioreactor of claim 3, further comprising at least one supporting member positioned outside the confined region of the chamber for supporting the second substrate.

5. The bioreactor of claim 3, further comprising at least one supporting member positioned inside the confined region of the chamber for supporting the second substrate.

6. A bioreactor comprising:

i. a first substrate having a first surface and an opposite second surface, defining a chamber therebetween the first surface and the opposite second surface of the first substrate for receiving biological cells and a liquid medium;

ii. an inlet port formed in the first substrate and apart from the chamber;

iii. a first connection channel formed in the first substrate, wherein the first connection channel is in fluid communication with the inlet port and the chamber for allowing a stream of substance to be delivered to the chamber;

iv. an outlet port formed in the first substrate and apart from the chamber;

v. a second connection channel formed in the first substrate, wherein the second connection channel is in fluid communication with the outlet port and the chamber for allowing a stream of substance to be removed from the chamber; and

vi. confining means positioned in the chamber to form a confinement region to confine the biological cells therein with the liquid medium, wherein the confining means comprises an array of posts,

wherein the chamber, the inlet portion, the first connection channel, the outlet port, and the second connection channel are all formed in the first substrate such that, in operation, the stream of substance flows from the inlet port through the first connection channel, the chamber and the second connection channel to the outlet port in a direction that is substantially parallel to the plane of the first surface of the first substrate; and

wherein the first substrate further defines a first alternate port and a third connection channel in fluid communication with the first alternate port and the confined region of the chamber for allowing seed biological cells to perfuse only inside the confined region in the chamber.

7. The bioreactor of claim 6, wherein the first substrate is fabricated from glass, Mylar, PDMS, silicon, a polymer, a semiconductor, or any combination of them.

8. The bioreactor of claim 6, further comprising a second substrate having a first surface and an opposite, second surface, wherein the second substrate is sized such that when the first surface of the second substrate is received by the second surface of the first substrate, the chamber is covered.

9. The bioreactor of claim 8, further comprising at least one supporting member positioned outside the confined region of the chamber for supporting the second substrate.

10. The bioreactor of claim 8, further comprising at least one supporting member positioned inside the confined region of the chamber for supporting the second substrate.

11. The bioreactor of claim 6, wherein the stream of substance is controlled so as to provide a gradient to the channel.

12. The bioreactor of claim 6, wherein the stream of substance comprises chemokine.

13. The bioreactor of claim 6, wherein the stream of substance comprises a substance affecting the growth of biological cells.

14. The bioreactor of claim 6, wherein sidewalls at the intersections of the first connection channel with the chamber and the second connection channel with the chamber are tapered to form an angle of inclination between about 10 and 45 degrees from vertical and an enclosed angle between about 30 and 80 degrees.