Systems, methods, and devices relating to a biomimetic cellularized nephron unit

• US 9,657,261 B2
• Filed: 06/15/2012
• Issued: 05/23/2017
• Est. Priority Date: 06/15/2011
• Status: Active Grant

• Current Assignee: Charles Stark Draper Laboratory Incorporated
• Original Assignee: Charles Stark Draper Laboratory Incorporated
• Inventors: Charest, Joseph L., Borenstein, Jeffrey T., Frohlich, Else
• Primary Examiner(s): JOHNSON, KARA D
• Application Number:
  US13/524,981
• Publication Number:
  US 20120318726A1
• Time in Patent Office: 1,803 Days
• Field of Search: 435395
• CPC Class Codes: A61M 1/14 Dialysis systems; Artificial kidneys; Blood oxygenators semi-permeable membranes characterised by the material, manufacturing processes therefor B01D71/00 ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis
• CPC Class Codes: C12M 23/00 Constructional details, e.g. recesses, hinges flow directing inserts in C12M27/18-C12M27/24; apparatus for chemical or physical processes in general B01J, chemical or physical laboratory apparatus in general B01L
• CPC Class Codes: C12M 23/06 Tubular C12M23/08, C12M23/16 take precedence
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 25/00 Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
• CPC Class Codes: C12M 25/10 Hollow fibers or tubes hollow fiber modules in general B01D63/02
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis

Abstract

Disclosed herein are systems and devices for culturing cells in a biomimetic environment of a cellularized nephron unit, and methods for fabricating and using the cellularized nephron unit.

Claims

1. A bioartificial kidney comprising:

a microfluidic flow channel comprising at least one topographical surface;

an inlet in fluid connection with the flow channel for allowing fluid to flow into the flow channel; and

renal cells seeded on the topographical surface as a confluent monolayer;

wherein

the topography of the surface of the flow channel comprises ridges parallel to a length of the microfluidic flow channel and having a width less than 5 μ

m and wherein the confluent monolayer of renal cells forms a substantially fluid-impermeable barrier.

2. The bioartificial kidney of claim 1, wherein the topography of the surface is configured to promote increased adhesion of cells in the cell layer to the at least one surface.

3. The bioartificial kidney of claim 1, further comprising a fluid source coupled to the flow channel via the inlet, wherein the fluid source is configured to flow a fluid through the flow channel and induce a shear stress upon the cell layer.

4. The bioartificial kidney of claim 3, wherein the flow channel is formed as part of one or more structures selected from the group consisting of a Loop of Henle, a collecting tubule and a distal tubule.

5. The bioartificial kidney of claim 3, wherein the fluid source is configured to flow the fluid at a flow rate that results in a level of shear stress on the cell layer that is less than or equal to about 10.0 dyne/cm2 in at least one region of the bioartificial kidney.

6. The bioartificial kidney of claim 3, wherein the fluid source is configured to flow the fluid at a flow rate that results in a level of shear stress on the cell layer that is less than or equal to about 1.0 dyne/cm2 in at least one region of the bioartificial kidney.

7. The bioartificial kidney of claim 3, wherein the fluid source is configured to flow the fluid at a flow rate that results in a level of shear stress on the cell layer that is less than or equal to about 0.1 dyne/cm2 in at least one region of the bioartificial kidney.

8. The bioartificial kidney of claim 3, wherein the fluid source is configured to flow the fluid at a flow rate that results in a level of shear stress on the cell layer that is about 0.02 dyne/cm2 in at least one region of the bioartificial kidney.

9. The bioartificial kidney of claim 3, wherein the flow channel is configured such that the fluid flows at a first flow rate in a first region of the bioartificial kidney and at a second flow rate in a second region of the bioartificial kidney.

10. The bioartificial kidney of claim 9, wherein the first flow rate results in a first level of shear stress on the cell layer in the first region, and wherein the second flow rate results in a second level of shear stress on the cell layer in the second region, and wherein the first and second levels of shear stress are different.

11. The bioartificial kidney of claim 9, wherein the bioartificial kidney comprises a Loop of Henle comprising an ascending limb and a descending limb, and wherein the first region is the ascending limb of the Loop of Henle, and the second region is the descending limb of the Loop of Henle.

12. The bioartificial kidney of claim 9, wherein the bioartificial kidney comprises a collecting duct, a distal tubule, and a Loop of Henle comprising an ascending limb and a descending limb, wherein the first region is in the Loop of Henle, and the second region is in one or more of a collecting duct and a distal tubule.

13. The bioartificial kidney of claim 1, further comprising a first and second surface, wherein the first surface of the flow channel has a first topography, and wherein the second surface of the flow channel has a second topography.

14. The bioartificial kidney of claim 13, wherein the bioartificial kidney comprises a Loop of Henle comprising an ascending limb and a descending limb, wherein the first surface is in the ascending limb of the Loop of Henle, and wherein the second surface is in the descending limb of the Loop of Henle.

15. The bioartificial kidney of claim 13, wherein the bioartificial kidney comprises a collecting duct, a distal tubule, and a Loop of Henle comprising an ascending limb and a descending limb wherein the first surface is in the Loop of Henle, and wherein the second surface is in one or more of a collecting duct and a distal tubule.

16. The bioartificial kidney of claim 13, further comprising a transition topography surface between the first and second surface.

17. The bioartificial kidney of claim 13, wherein the topography of the first surface comprises ridges with a first pitch, and the topography of the second surface comprises ridges with a second pitch.

18. The bioartificial kidney of claim 13, wherein the topography of the first surface comprises ridges in a first orientation with respect to fluid flow, and the topography of the second surface comprises ridges in a second orientation with respect to fluid flow.

19. The bioartificial kidney of claim 13, wherein the first topography comprises ridges, and the second topography comprises one or more topographies of the pit or post families.

20. The bioartificial kidney of claim 1, further comprising a cytophilic substance disposed on a portion of a substrate for growing the cell layer in the portion of the substrate, and wherein the portion of the substrate forms a surface of the flow channel.

21. The bioartificial kidney of claim 20, wherein the cytophilic substance comprises a collagen protein.

22. The bioartificial kidney of claim 1, wherein the surface topography is configured to cause the arrangement, behavior, or morphology of the cell layer to replicate an arrangement, behavior, or morphology of cells in a kidney.

23. The bioartificial kidney of claim 1, further comprising at least a second flow channel having at least one surface of the second flow channel having a topography formed therein.

24. The bioartificial kidney of claim 23, wherein the first flow channel is separated from the second flow channel by a membrane.

25. The bioartificial kidney of claim 23, wherein the first flow channel is seeded with renal epithelial cells.

26. The bioartificial kidney of claim 23, wherein the second flow channel is seeded with vascular epithelial cells.

27. The bioartificial kidney of claim 23, wherein the first flow channel comprises a blood flow layer, and the second flow channel comprises a filtrate layer.

28. The bioartificial kidney of claim 23, wherein at least one surface of the first channel and includes a different surface topography than a corresponding surface in the second channel.

29. The bioartificial kidney of claim 28, wherein the surface topography in the first channel comprises a different pitch or shape than the surface topography in the second channel.

30. The bioartificial kidney of claim 23, further comprising a first fluid source for flowing a fluid through the first flow channel and the second flow channel, wherein the fluid induces a first shear stress upon the cell layer in the first channel and a second shear stress upon the cell layer in the second channel.

31. The bioartificial kidney of claim 30, wherein the first shear stress is different than the second shear stress.

32. The bioartificial kidney of claim 1, wherein the renal cells comprise one or more cell types selected from the group consisting of proximal tubule cells, renal proximal tubule epithelial cells, Madin-Darby canine kidney cells, primary inner medullary collecting duct cells, primary proximal tubule cells, embryonic stem-cells, adult stem-cells, induced pluripotent stem cells and endothelial cells.

33. A bioartificial kidney comprising:

a microfluidic flow channel comprising at least one topographical surface;

an inlet in fluid connection with the flow channel for allowing fluid to flow into the flow channel; and

renal cells seeded on the topographical surface;

wherein the topography of the surface of the flow channel comprises ridges parallel to a length of the microfluidic flow channel and having a width of about 1 μ

m or about 3 μ

m and is selected to cause the renal cells to form a cell layer disposed on the surface to achieve an arrangement, behavior, or morphology determined at least in part by the topography of the at least one surface.