Pumpless microfluidic organ-on-a-chip system including a functional immune system

Patent metadata

Field	Value
Publication number	US-11554373-B2
Application number	US-201716474768-A
Country	US
Kind code	B2
Filing date	Dec 29, 2017
Priority date	Dec 30, 2016
Publication date	Jan 17, 2023
Grant date	Jan 17, 2023

Abstract

A pumpless microfluidic system is disclosed that can be used to mimic the interaction of organ systems with the immune system. Also disclosed is a method for mimicking an immune system, comprising culturing a plurality of organ cells and at least one population of immune cells in the disclosed pumpless microfluidic system under physiological conditions. The method can further comprise activating an immune reaction in the pumpless microfluidic system, continuing the culture for a defined period, collecting a sample of culture medium from the system, and assaying the sample for one or more indicators of an immune response.

First claim

What is claimed is:

1. A gravity driven pumpless microfluidic system, comprising:
a first chamber containing a first plurality of organ cells attached to at least a portion of a cell attachment surface of the first chamber, wherein the first plurality of organ cells mimics a function of a first organ;
a microcantilever sensor to non-invasively measure a function of the first plurality of organ cells; and
at least one population of living immune cells, including at least one population of leukocytes, recirculating through the first chamber in a serum free culture medium for at least two days without activation,
wherein the serum free medium comprises GDNF, BDNF, CNTF, NT3, NT4, vitronectin, agrin, sonic hedgehog, laminin, cAMP, retinoic acid, IGF-1, and NaCl.

2. The pumpless microfluidic system of claim 1 , further comprising a second chamber in fluid connection with the first chamber containing a second plurality of organ cells attached to at least a portion of a cell attachment surface of the second chamber, wherein the second plurality of organ cells mimic a function of a second organ.

3. The pumpless microfluidic system of claim 2 , further comprising a third chamber in fluid connection with the first and second chambers containing a third plurality of organ cells attached to at least a portion of a cell attachment surface of the third chamber, wherein the third plurality of organ cells mimic a function of a third organ.

4. The pumpless microfluidic system of claim 1 , wherein the organ cells are selected from the group consisting of: cardiomyocytes, skeletal muscle myotubes, hepatocytes, kidney cells, neurons, epithelial cells, astrocytes, Schwann cells, bone marrow cells, cancer cells lines (drug-resistant and non-drug resistant), blood vessel endothelial cells, pancreatic islet cells, oligodendrocytes, synoviocytes, and fibroblasts.

5. The pumpless microfluidic system of claim 1 , wherein first chamber and the microcantilever sensor are arranged on a chip.

6. The pumpless microfluidic system of claim 1 , further comprising a recording device operably connected to the microcantilever sensor, wherein the recording device is configured to record data from the microcantilever sensor.

7. The pumpless microfluidic system of claim 6 , wherein the first chamber is one of a plurality of chambers, and the system further comprises one or more microfluidic channels interconnecting the plurality of chambers.

8. The pumpless microfluidic system of claim 1 , wherein the at least one population of immune cells comprise at least one of: peripheral blood mononuclear cells (PBMCs); granulocytes selected from the group consisting of neutrophils, eosinophils, basophils, and mast cells; or agranulocytes selected from the group consisting of monocytes, macrophages, lymphocytes, and natural killer (NK) cells.

9. The pumpless microfluidic system of claim 1 , further comprising at least one agonist or antagonist of an innate immune response.

10. The pumpless microfluidic system of claim 9 , wherein agonist or antagonist of the innate immune response comprises a complement protein or an interferon and/or activates a complement cascade.

11. The pumpless microfluidic system of claim 1 , further comprising at least one agonist or antagonist of an adaptive immune response.

12. The pumpless microfluidic system of claim 11 , wherein agonist or antagonist of the adaptive immune response comprises an antibody or a cytokine.

13. A method for mimicking an immune system, the method comprising;
attaching a first plurality of organ cells to at least a portion of a cell attachment surface in a first chamber of a gravity driven pumpless microfluidic system;
measuring a function of the first plurality of organ cells with a microcantilever sensor;
recirculating at least one population of immune cells, including at least one population of leukocytes, through the first chamber in a serum free culture medium comprising GDNF, BDNF, CNTF, NT3, NT4, vitronectin, agrin, sonic hedgehog, laminin, cAMP, retinoic acid, IGF-1, and NaCl; and
maintaining viability of the at least one population of leukocytes for at least two days without activation.

14. The method of claim 13 , further comprising evaluating the system for an immune response.

15. The method of claim 14 , wherein the immune response is evaluated by monitoring the first plurality of organ cells for immune cell infiltration.

16. The method of claim 14 , further comprising activating an immune reaction in the pumpless microfluidic system and continuing the culture for a defined period prior to evaluating the system for an immune response.

17. The method of claim 16 , wherein the immune reaction is activated by a physical insult to the first plurality of organ cells or by adding one or more chemical or biological agents to the culture medium.

18. The method of claim 13 , further comprising measuring a functional readout of tissue damage.

Assignees

• Univ Central Florida Res Found Inc
• Hesperos Inc

Inventors

• Hickman James
• Colon Alisha
• Mcaleer Christopher
• Sasserath Trevor
• Rumsey John
• Elbrecht Daniel

Classifications

• B01L2300/0877

  Flow chambers · CPC title

• C12M23/16 Primary

  Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093) · CPC title

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title

• B01L3/5027

  by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip · CPC title

• B01L3/502753 Primary

  characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation · CPC title