PERIFUSION DEVICE

• US 20110020856A1
• Filed: 06/30/2010
• Published: 01/27/2011
• Est. Priority Date: 01/12/2007
• Status: Active Grant

• Current Assignee: Biorep Technologies, Inc., University of Miami
• Original Assignee: Biorep Technologies, Inc., University of Miami
• Inventors: BERGGREN, PER-OLOF, POO, RAMON E., Cabrera, Over, ECHEVERRI, FELIPE, RICORDI, CAMILLO
• Granted Patent: US 8,263,389 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/40 Manifolds; Distribution pieces fluid transfer means B01L3/563
• CPC Class Codes: C12M 23/42 Integrated assemblies, e.g. cassettes or cartridges
• CPC Class Codes: C12M 41/00 Means for regulation, monitoring, measurement or control, e.g. flow regulation controlling or regulating chemical, physical or physicochemical processes B01J19/0006; heating or cooling apparatus for laboratory use B01L7/00; electro optical investigation of individual particles, flow cytometers G01N15/14; automatic analysis G01N35/00; controlling or regulating in general G06N
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: Y10T 137/87249 Multiple inlet with multiple outlet

Abstract

A perifusion device includes at least one sample container for cells, the sample container having an inlet and an outlet. The container receives test liquid through the inlet and discharges the liquid through the outlet. A manifold having a plurality of liquid inlets, control valves, and liquid outlets can be provided so that the flow of liquids from source containers to the sample containers can be varied and controlled. A receptacle housing has a plurality of receptacles for receiving fluid from the outlet of the sample container. A drive is connected to the receptacle housing for moving the receptacle housing such that liquid samples are collected sequentially from the outlet of the sample containers. A programmable controller can be provided to control movement of the receptacle housing at predetermined times, and to record data identifying liquid samples in the receptacles. The test liquid includes at least one stimuli for the cells, which can be the presence, absence, or concentration of a compound in the liquid, or a physical property of the liquid such as temperature. The liquid collected in the receptacles is analyzed to determine the response of the cells to the stimuli.

Claims

1. A perifusion device, comprising:

a plurality of sample containers;

the sample containers having a liquid inlet and a liquid outlet, the containers receiving liquid through the inlet and discharging through the liquid outlet;

a receptacle housing having a plurality of receptacles for receiving liquid from the outlets of the sample containers, said plurality of receptacles being positioned in said receptacle housing such that different receptacles receive samples from different liquid outlets of said sample containers;

a drive connected to the receptacle housing for moving said receptacle housing such that samples from the liquid outlets of said sample containers are successively collected in different ones of said plurality of receptacles;

a valve manifold having a plurality of liquid inlets and a plurality of liquid outlets, and a plurality of flow junctions connecting said liquid inlets to said liquid outlets, and at least one valve disposed between each liquid inlet and each flow junction, said liquid outlets of said valve manifold being connected to said liquid inlets of said sample containers;

a plurality of liquid sources for supplying liquid to said liquid inlets, each liquid source being in liquid connection to one of said liquid inlets of said valve manifold; and

,

a programmable controller for controlling the operation of said valves and the flow of liquid from said liquid sources to the mixing sites, and thereby to the sample containers.

2. The perifusion device of claim 1, wherein each of said liquid inlets is connected to a liquid inlet conduit of said manifold, said liquid inlet conduit having a plurality of branch lines, each branch line being connected to at least one of said flow junctions and having a valve associated with said branch line to control liquid flow through said branch line and to said flow junction.

3. The perifusion device of claim 2, wherein each flow junction connects to a plurality of liquid outlets of said valve manifold.

4. The perifusion device of claim 2, wherein each flow junction connects to a plurality of said branch lines and to a plurality of liquid inlets and liquid sources.

5. The perifusion device of claim 1, wherein said receptacles are arranged in rows and columns and said liquid outlets of said sample containers are aligned in a row, said rows of receptacles being aligned with said row of liquid outlets to receive samples from said sample containers, movement of said receptacle housing by said drive causing successive rows of receptacles in said columns to receive successive samples from said sample containers.

6. The perifusion device of claim 1, wherein said receptacle housing is a tray, said tray being detachably securable to a receptacle drive assembly in either landscape or portrait positions.

7. The perifusion device of claim 1, further comprising a heater for heating gas flowing through said perifusion device, said heater comprising a plurality of gas inlets and a plurality of gas outlets.

8. The perifusion device of claim 1, wherein said receptacles are wells.

9. The perifusion device of claim 1, wherein said sample containers are columns.

10. The perifusion device of claim 1, wherein said sample containers are packed with substrate.

11. The perifusion device of claim 10, wherein said substrate comprises at least one selected from the group consisting of beads and gel.

12. The perifusion device of claim 1, further comprising a pump for causing said liquid to flow through said column.

13. The perifusion device of claim 12, wherein said pump controls the volumetric flow rate through said container.

14. The perifusion device of claim 12, wherein said pump is a peristaltic pump.

15. The perifusion device of claim 1, further comprising a programmable controller for operating said drive to move said receptacle housing at predetermined times.

16. The perifusion device of claim 15, wherein said programmable controller is a computer.

17. The perifusion device of claim 1, further comprising at least one sensor for sensing a characteristic of the liquid.

18. The perifusion device of claim 17, comprising at least one sensor upstream of the sample and at least one sensor downstream of the sample.

19. The perifusion device of claim 1, wherein said liquid comprises a cell stimulus compound, said compound being at least one selected from the group consisting of carbohydrate, lipid, and peptide.

20. The perifusion device of claim 1, wherein said cell stimulus compound comprises a drug which stimulates cell behavior in some detectable way.

21. The perifusion device of claim 1, further comprising at least one temperature controller for controlling the temperature of said liquid flowing through said sample container.

22. The perifusion device of claim 1, comprising first and second sample containers, the output of said first sample containers flowing into and becoming the input for said second sample containers.

23. A method for performing perifusion, comprising the steps of:

providing a perifusion device, said perifusion device comprising a plurality of sample containers having a liquid inlet and a liquid outlet, the containers receiving liquid through the inlet and discharging through the outlet;

a receptacle housing having a plurality of receptacles for receiving liquid from the outlet of the sample container, said plurality of receptacles being positioned in said receptacle housing such that different receptacles receive samples from different liquid outlets of said sample containers;

a drive connected to the receptacle housing for moving said receptacle housing such that samples from the liquid outlets of said sample containers are successively collected in different ones of said plurality of receptacles;

a valve manifold having a plurality of liquid inlets and a plurality of liquid outlets, and a plurality of flow junctions connecting said liquid inlets to said liquid outlets, and at least one valve disposed between each liquid inlet and each mixing site;

a plurality of liquid sources for supplying liquid to said containers, each liquid source being connected to one of said liquid inlets of said valve manifold;

said liquid outlets of said valve manifold being connected to said liquid inlets of said sample containers; and

, a programmable controller for controlling the operation of said valves and the flow of liquid from said liquid sources to the mixing sites, and thereby to the sample containers;

placing sample cells into said containers;

providing in said liquid sources at least one stimulus for said cells;

causing said liquid containing said stimulus to flow through said valve manifold;

operating said control means to cause said valves to control the flow of liquids from said liquid sources to said flow junctions, said liquids being mixed in said flow junctions and exiting through said liquid outlets to said inlets of said sample containers;

collecting said liquid from said liquid outlets of said sample containers in said receptacles;

causing said drive to move said receptacle housing;

collecting at least one additional sample in at least one additional receptacle; and

,

detecting the response of the cells to the stimulus in said collected samples.

24. A device for conducting perifusion, comprising:

first and second liquid source containers, said source containers having differing concentrations of a substance;

a pump connected to each source container for metering determined flow rates from each source container;

a flow junction for combining the flows from each source container, the output from the flow junction being directed to at least one sample container;

a receptacle housing having a plurality of receptacles for receiving liquid from outlets of the sample container, said plurality of receptacles being positioned in said receptacle housing such that different receptacles sequentially receive liquid output from the outlet of said sample container;

a drive connected to the receptacle housing for moving said receptacle housing such that samples from the liquid outlet of said sample container are successively collected in different ones of said plurality of receptacles; and

,

a programmable controller for controlling the operation of said pumps and the combining of said flows to obtain a desired final concentration of said substance flowing into the sample container.