MINIATURIZED, AUTOMATED IN-VITRO TISSUE BIOREACTOR

• US 20140322701A1
• Filed: 04/29/2014
• Published: 10/30/2014
• Est. Priority Date: 04/30/2013
• Status: Active Grant

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Original Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Inventors: Pannu, Satinderpall S., Tooker, Angela C., Wheeler, Elizabeth K., Sheth, Heeral, Chen, Haiyin, Hudson, Bryan D., Kulp, Kris, Mcnerney, Margaret W., Qian, Fang
• Granted Patent: US 10,793,820 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 41/26 of pH
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00

Abstract

In one embodiment, a system includes a bioreactor coupled to a substrate. The bioreactor includes: a plurality of walls defining a reservoir; a plurality of fluidic channels in at least some of the walls; a fluidic inlet in fluidic communication with the reservoir via, the fluidic channels; a fluidic outlet in fluidic communication with the reservoir via the fluidic channels; and one or more sensors coupled to the reservoir, each sensor being configured to detect one or more of: environmental conditions in the reservoir; and physiological conditions of one or more cells optionally present in the reservoir. In another embodiment, a method includes delivering media to a reservoir of a bioreactor; delivering a, plurality of cells to the reservoir, controlling a reservoir temperature and a reservoir gas composition; using one or more of the sensors to monitor environmental and physiological conditions; and reporting the environmental and/or physiological conditions.

Claims

1. A system, comprising:

a bioreactor coupled to a substrate, the bioreactor comprising;

a plurality of walls defining a reservoir;

a plurality of fluidic channels in at least some of the walls;

a fluidic inlet in fluidic communication with the reservoir via the fluidic channels;

a fluidic outlet in fluidic communication with the reservoir via the fluidic channels; and

one or more sensors coupled to the reservoir, each sensor being configured to detect one or more of;

environmental conditions in the reservoir; and

physiological conditions of one or more cells optionally present in the reservoir.

2. The system as recited in claim 1, wherein the sensors comprise one or more of:

chemical sensors configured to detect chemical conditions in the reservoir;

mechanical sensors configured to detect one or more mechanical forces acting on contents of the reservoir;

electrical sensors configured to detect one or more electrical forces acting on contents of the reservoir; and

sensors comprising a material selected from the group consisting of;

platinum, indium oxide, gold and activated iridium oxide.

3. The system as recited in claim 2, wherein at least one surface of the sensors is functionalized to detect one or more of:

biological markers, oxidative stress, cytotoxicity, neurotransmitters, reservoir pH, osmolality, cellular metabolism, an expression level of one or more proteins of interest, an expression level of one or more DNA sequences, and an expression level of one or more mRNA sequences.

4. The system as recited in claim 2, wherein the sensors are arranged in a three-dimensional pattern.

5. The system as recited in claim 1, further comprising a control board configured to facilitate communication between the bioreactor and a processor.

6. A system, comprising:

a substrate;

a plurality of the bioreactors as recited in claim 1 coupled to the substrate; and

a control board configured to;

facilitate communication between the system and a processor; and

independently control each of the bioreactors.

7. The system as recited in claim 1, further comprising:

a plurality of openings in one of the walls, the openings configured to facilitate;

adding one or more of cells and reagents to the reservoir, and

removing one or more of cells and reagents from the reservoir; and

transferring fluid between the reservoir and another reservoir

a plurality of plugs, each plug being adapted to engage one of the openings and selectively control gas exchange and material exchange between the reservoir and an environment external to the reservoir.

8. The system as recited in claim 1, wherein the fluidic inlet, the fluidic outlet, and the reservoir are configured to mix one or more fluids in the reservoir.

9. The system as recited in claim 1, wherein the substrate comprises a glass layer and a polymer layer.

10. The system as recited in claim 1, wherein the substrate comprises an optically transparent viewing window configured to transmit optical information from the reservoir.

11. The system as recited in claim 1, further comprising a vacuum pump in communication with the reservoir via a plurality of channels in the substrate.

12. The system as recited in claim 1, further comprising:

a gas-source in communication with the reservoir via a plurality of channels in at least one of the walls.

13. The system as recited in claim 1, wherein the walls comprise a material selected from a group consisting of:

dielectrics, silicon-based polymers, polydimethylsiloxane (PDMS), polystyrene, spin-on-glass, and polyurethane.

14. The system as recited in claim 1, further comprising a heat source in thermal communication with the reservoir, the heat source comprising one or more of:

a resistive heater embedded in the substrate; and

a heating chamber comprising temperature control, the heating chamber surrounding the bioreactor.

15. The system as recited in claim 1, further comprising one or more environmental controls.

16. The system as recited in claim 1, further comprising integrated electronics configured to perform environmental control, fluidic processing and control, sensing and stimulating cells functionalities.

17. The system as recited in claim 1, wherein the system is compatible with conventional microbiology techniques to allow traditional assays and robotic manipulations.

18. The system as recited in claim 1, wherein the system is optically transparent to allow for optical assays.

19. The system as recited in claim 1, wherein the system is configured to connect to other bio-reactors to recapitulate additional functions.

20. The system as recited in claim 1, further comprising one or more three-dimensional tissue constructs in the bio-reactor.

21. A method, comprising:

delivering media to a reservoir of a bioreactor;

delivering a plurality of cells to the reservoir, wherein at least some of the cells are disposed onto one or more sensors in the bioreactor;

controlling a reservoir temperature and a reservoir gas composition;

using one or more of the sensors to monitor one or more of;

environmental conditions in the reservoir, and

physiological conditions of at least some of the cells; and

communicating the monitored environmental conditions and/or physiological conditions to an external device.

22. The method as recited in claim 21, further comprising:

evacuating the media from the reservoir via a fluidic outlet in fluidic communication with the reservoir to either remove waste or for further analysis; and

delivering new media, reagents or cells to the reservoir.

23. The method as recited in claim 21, further comprising:

stimulating at least some of the cells using the sensors,

monitoring the stimulated cells to detect a physiological response to the stimulation; and

communicating any detected physiological response to the external device.

24. The method as recited in claim 21, further comprising autoclaving the bioreactor, wherein the autoclaving does not detrimentally affect operation of any electrical component of the bioreactor.

25. The method as recited in claim 21, wherein at least some of the sensors comprise a plurality of nanowire sensors arranged in a three-dimensional matrix throughout the reservoir, and

wherein monitoring one or more of the environmental conditions and the physiological conditions is performed in three dimensions using the plurality of nanowire sensors.