Capillary Perfused Bioreactors with Multiple Chambers

• US 20090215654A1
• Filed: 03/05/2009
• Published: 08/27/2009
• Est. Priority Date: 08/27/2002
• Status: Active Grant

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Wikswo, John P., Lin, Charles P., Hofmeister, William H., Leboeuf, Eugene J., Prokop, Ales, Reiserer, Randall S., Chung, Chang Y., Cliffel, David, Haselton, Frederick R., Stremler, Mark A., McCawley, Lisa J., Baudenbacher, Franz J.
• Granted Patent: US 8,003,378 B2
• Time in Patent Office: Days
• US Class Current: 506/33
• 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, an opposite second surface and edges;

ii. a second substrate having a first surface and an opposite second surface, defining a cavity with a bottom surface, wherein the bottom surface is located therebetween the first surface and the second surface, and wherein 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 biological cells and a liquid medium;

iii. a recess formed in the second substrate with a bottom surface and in fluid communication with the channel; and

iv. a barrier positioned for covering the recess so as to form an outer chamber, wherein the barrier has a porosity to allow the channel and the outer chamber in fluid communication and control the move of at least one predetermined type of biological cells between the channel and the outer chamber.

2. The bioreactor of claim 1, wherein the second substrate further defines a first opening and an opposite, second opening adapted for allowing a flow of liquid to be introduced into the channel through the first opening and away from the channel through the second opening substantially along a first direction.

3. The bioreactor of claim 2, further comprising a biocompatible coating layer applied to the interior surfaces of the second substrate around the channel.

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28. A bioreactor comprising:

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

ii. an inlet port formed in the first substrate;

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;

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 a region in the chamber proximate to the first connection channel to confine the biological cells.

29. The bioreactor of claim 28, wherein the confining means comprises a plurality of traps, wherein each of the plurality of traps is capable of receiving at least one biological cell.

30. The bioreactor of claim 29, wherein each of the plurality of traps comprises a structure defining a recess so as to receive and confine at least one biological cell therein.

31. The bioreactor of claim 30, wherein the structure is partially formed with a filter to allow the recess to be in fluid communication with the chamber.

32. The bioreactor of claim 29, wherein the plurality of traps forms an array.

33. The bioreactor of claim 28, 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 first connection channel for allowing additional substance to be introduced into the chamber.

34. The bioreactor of claim 28, wherein the first substrate further defines a second alternate port, a third connection channel, and a second chamber, wherein the third connection channel is in fluid communication with second alternate port and the second chamber, and the second chamber is in fluid communication with the first connection channel.

35. The bioreactor of claim 34, wherein the second chamber is formed with an oxygen permeable structure to provide oxygen to the biological cells.

36. The bioreactor of claim 28, wherein the first substrate is fabricated from glass, Mylar, PDMS, silicon, a polymer, a semiconductor, or any combination of them.

37. The bioreactor of claim 28, further comprising a second substrate having a first surface and an opposite, second surface, and means adapted for electrochemical measurements of the biological cells in the chamber, wherein the means for electrochemical measurements is positioned with the second substrate such that when the first surface of the second substrate is received by the second surface of the first substrate, the means for electrochemical measurements is at a corresponding measurement position.

38. The bioreactor of claim 37, wherein the means for electrochemical measurements comprises:

i. at least one electrode monitoring entry of the biological cells into the chamber;

ii. at least one electrode monitoring leaving of the biological cells from the chamber; and

iii. a plurality of electrodes detecting chemical species in the chamber.

39. The bioreactor of claim 28, further comprising a third substrate having a first surface and an opposite, second surface, and means adapted for optical measurements, wherein the means for optical measurements is positioned with the third substrate such that when the first surface of the third substrate is received by the second surface of the first substrate, the means for optical measurements is at a corresponding measurement position.

40. The bioreactor of claim 39, wherein the means for optical measurements comprises a plurality of optical sensors strategically positioned for detecting chemical and biological species within the chamber and the physiological state of the biological cells within the chamber.

41. The bioreactor of claim 39, wherein the third substrate is at least partially transparent.

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50. A bioreactor comprising:

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

ii. at least one second chamber formed in the first substrate for receiving a second type of biological cells and a liquid medium;

iii. at least one connection channel formed in the first substrate, wherein at least one connection channel is in fluid communication with a corresponding second chamber and the first chamber for allowing the first type of biological cells and the second type of the biological cells to interact with each other.

51. The bioreactor of claim 50, wherein the first type of biological cells comprises protozoa, and the second type of biological cells comprises bacteria.

53. The bioreactor of claim 51, wherein the connection channel is formed to allow protozoa to travel therein, and further comprising a sizing limiting post to limit the mobility of protozoa.

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