Devices and methods for pharmacokinetic-based cell culture system

• US 8,030,061 B2
• Filed: 09/08/2006
• Issued: 10/04/2011
• Est. Priority Date: 04/25/2001
• Status: Expired due to Fees

• Current Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Original Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Harrison, Robert Andrew, Shuler, Michael, Meyers, Scott, Baxter, Gregory T., Sin, Aaron
• Primary Examiner(s): Beisner; William H
• Application Number:
  US11/530,381
• Publication Number:
  US 20070020613A1
• Time in Patent Office: 1,852 Days
• Field of Search: 435/373, 435/288.5, 435/293.1, 435/294.1
• US Class Current: 435/288.5
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture
• 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
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0671 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5067 Liver cells

Abstract

Devices, in vitro cell cultures, systems, and methods are provided for microscale cell culture analogous (CCA) device.

Claims

1. A pharmacokinetic-based microscale culture device, comprising:

a first microscale chamber containing a first type of cell, wherein the first microscale chamber is dimensioned to maintain the first type of cell under conditions that give rise to least one pharmacokinetic parameter value in vitro that is comparable to a value for the same at least one pharmacokinetic parameter obtained with respect to the same type of cell in vivo, wherein the at least one pharmacokinetic parameter comprises metabolism by cells, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of culture medium, and wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio;

a second microscale chamber containing a second type of cell, wherein the second microscale chamber is dimensioned to maintain the second type of cell under conditions that give rise to at least a second pharmacokinetic parameter value in vitro comparable to a value for the same at least a second pharmacokinetic parameter obtained with respect to the same second type of cell in vivo, wherein the second microscale chamber comprises a second inlet and a second outlet for flow of culture medium; and

a microfluidic channel interconnecting the first and second microscale chambers wherein the microfluidic channel is dimensioned to transport a culture medium, and wherein the microscale culture device is dimensioned to maintain at least one desired value for shear stress under a condition of flow of the culture medium.

6. The device of claim 1, 2, 3, 4, or 5, further comprising a debubbler configured to remove bubbles from fluid that flows in or through the device.

7. The device of claim 6 wherein the debubbler is located within the device.

8. The device of claim 6 wherein the debubbler is located externally to the device.

9. The device of claim 1, 2, 3, 4, or 5, wherein said first type of cell comprises a cell from heart, kidney, stomach, intestine, lung, liver, skin, fat, bone, cartilage, skeletal muscle, smooth muscle, cardiac muscle, bone marrow, muscle, brain, or pancreas.

10. The device of claim 1, 3, or 5, wherein the second type of cell comprises malignant or diseased cells.

11. The device of claim 1, 3, or 5, wherein one of the microscale chambers contains a tumor biopsy.

12. The device of claim 1, 3, or 5, wherein the first and second type of cell are selected to effect the metabolism of a drug.

13. The device of claim 1, 2, 3, 4, or 5, further comprising a sensor for obtaining signals from cultured cells.

14. The device of claim 13 wherein the sensor comprises a biosensor or a waveguide.

15. The device of claim 1, 2, 3, 4, or 5, wherein the first type of cell comprises liver cells.

16. The device of claim 1, 3, or 5, wherein at least one of the first and second type of cell is cultured on collagen or poly-lysine.

17. The device of claim 1, 3, or 5, wherein the first and second type of cells are, independently, circulating, immobilized, or adhered to the device through a biological material.

18. The device of claim 17, wherein said first type of cell and/or said second type of cell is selected from the group consisting of healthy tissue, diseased tissue, a portion of a tissue biopsy, a portion of skin, a portion of tissue, a portion of an artery, a portion of a vein, a portion of a gastrointestinal tract, a portion of an esophagus, a portion of a colon, a portion of a liver, a portion of an organ, a portion of a heart, a portion of a brain, a portion of a kidney, a portion of a lung, a portion of a muscle, a cell culture, an eukaryotic cell, a plant cell, an animal cell, a mammalian cell, a prokaryotic cell, a primary cell, a tumor cell, a stem cell, a genetically altered cell, a transformed cell, and an immortalized cell.

19. The device of claim 1, 2, 3, 4, or 5, wherein the device is further configured to provide a quantitative output.

20. The device of claim 1, 2, 3, 4, or 5, further comprising at least one additional type of cell.

21. The device of claim 1, 3, or 5, wherein the first and second type of cell are the same.

22. The device of claim 1, wherein the at least a second pharmacokinetic parameter comprises metabolism by cells.

23. The device of claim 1, 2, 3, 4, or 5, further comprising a pumping mechanism.

24. The device of claim 23, wherein the pumping mechanism is integrated in the device.

25. The device of claim 23, wherein the pumping mechanism is external to the device.

26. The device of claim 1, 3, or 5, wherein the first and/or second type of cell comprises a circulating or adherent cell.

27. The device of claim 1, 2, 3, 4, or 5, wherein said first type of cell comprises a cell from a tissue biopsy.

28. The device of claim 1, 2, 3, 4, or 5, wherein the device is microfabricated.

29. The device of claim 1, 2, 3, 4, or 5, wherein the device is manufactured from a microfabricated master.

30. The device of claim 1, 2, 3, 4, or 5, wherein the first and/or second type of cell is selected from the group consisting of a portion of healthy tissue, a portion of diseased tissue, a portion of a tissue biopsy, a portion of skin, a portion of tissue, a portion of an artery, a portion of a vein, a portion of a gastrointestinal tract, a portion of an esophagus, a portion of a colon, a portion of a liver, a portion of an organ, a portion of a heart, a portion of a brain, a portion of a kidney, a portion of a lung, a portion of a muscle, a cell culture, an eukaryotic cell, a plant cell, an animal cell, a mammalian cell, a prokaryotic cell, a primary cell, a tumor cell, a stem cell, a genetically altered cell, a transformed cell, and an immortalized cell.

31. The device of claim 1, 3, or 5, wherein the first and second microscale chambers are separated by a barrier.

32. The device of claim 31, wherein the barrier mimics a gastrointestinal barrier or a blood brain barrier.

2. A pharmacokinetic-based microscale culture device comprising:

a microscale chamber containing a first type of cell, wherein said microscale chamber is dimensioned to maintain the first type of cell under conditions that give rise to a value for at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter obtained with respect to the same type of cell in vivo, wherein the pharmacokinetic parameter is selected from the group consisting of liquid residence time and liquid to cell volume ratio, wherein the microscale chamber comprises a first inlet and a first outlet for flow of culture medium.

33. The device of claim 2 or 4, wherein the device is configured for the culture medium to flow re-circulated through the microscale chamber.

34. The device of claim 2 or 4, wherein the first type of cell is cultured on collagen or poly-lysine.

35. The device of claim 2 or 4, wherein the first type of cell is circulating, immobilized, or adhered to the device through a biological material.

36. The device of claim 35, wherein said first type of cell is selected from the group consisting of healthy tissue, diseased tissue, a portion of a tissue biopsy, a portion of skin, a portion of tissue, a portion of an artery, a portion of a vein, a portion of a gastrointestinal tract, a portion of an esophagus, a portion of a colon, a portion of a liver, a portion of an organ, a portion of a heart, a portion of a brain, a portion of a kidney, a portion of a lung, a portion of a muscle, a cell culture, an eukaryotic cell, a plant cell, an animal cell, a mammalian cell, a prokaryotic cell, a primary cell, a tumor cell, a stem cell, a genetically altered cell, a transformed cell, and an immortalized cell.

37. The device of claim 2 or 4, wherein the first type of cell comprises a circulating or adherent cell.

3. A pharmacokinetic-based microscale culture device comprising:

a first microscale chamber containing a first type of cell, wherein the first microscale chamber is dimensioned to maintain the first type of cell under conditions that give rise to a value for at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter obtained with respect to the same type of cell in vivo, wherein the pharmacokinetic parameter is selected from the group consisting of liquid residence time and liquid to cell volume ratio, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of culture medium, and

a second microscale chamber containing a second type of cell, wherein the second microscale chamber comprises a second inlet and a second outlet for flow of culture medium; and

a microfluidic channel interconnecting the first and second microscale chambers wherein the microfluidic channel is dimensioned to transport culture medium.

4. A pharmacokinetic-based microscale culture device comprising:

a microscale chamber containing a first type of cell, wherein said chamber is dimensioned to maintain the first type of cell under conditions that give rise to a value for at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter obtained with respect to the same type of cell in vivo, wherein the pharmacokinetic parameter is metabolism by cells, wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio, wherein the chamber comprises a first inlet and a first outlet for flow of culture medium.

5. A pharmacokinetic-based microscale culture device comprising:

a first microscale chamber containing a first type of cell, wherein the first microscale chamber is dimensioned to maintain the first type of cell under conditions that give rise to a value for at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter obtained with respect to the same type of cell in vivo, wherein the pharmacokinetic parameter is metabolism by cells, wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of culture medium, and;

a second microscale chamber containing a second type of cell, wherein the second chamber comprises a second inlet and a second outlet for flow of culture medium; and

a microfluidic channel interconnecting the first and second microscale chambers wherein the microfluidic channel is dimensioned to transport culture medium.

38. A method of culturing cells in a microscale culture device to provide a pharmacokinetic parameter in vitro comparable to that found in vivo comprising:

maintaining a first type of cell in a first microscale chamber in the microscale culture device, wherein said first microscale chamber is dimensioned to provide a value of at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter obtained with respect to the same type of cell in vivo, wherein the pharmacokinetic parameter is metabolism by cells, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of culture medium, and wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio;

maintaining a second type of cell in a second microscale chamber, wherein said second microscale chamber is dimensioned to provide a value of at least a second pharmacokinetic parameter in vitro that is comparable to a value for the same at least a second pharmacokinetic parameter obtained with respect to the same second type of cell in vivo, wherein the second microscale chamber comprises a second inlet and a second outlet for flow of culture medium;

and wherein the first and second microscale chambers are interconnected with a microfluidic channel wherein the microfluidic channel is dimensioned to transport the culture medium; and

, wherein the microscale culture device is dimensioned to maintain at least one desired value for shear stress under a condition of flow of the culture medium, and

culturing said first and said second types of cells in said microscale culture device to provide values for metabolism by said first type of cell comparable to in vivo values for metabolism by said first type of cell.

39. The method of claim 38, further comprising removing bubbles from fluid that flows in or through said chambers.

40. The method of claim 38 further comprising contacting the first type of cell with a material to be tested and evaluating the effect of said material.

41. The method of claim 40 wherein said evaluating is performed with a biosensor.

42. The method of claim 40 wherein said material comprises a drug, a cytotoxic substance, a potentially cytotoxic substance, a pharmaceutical composition or an inhibitory compound.

43. The method of claim 42 wherein the material comprises a drug.

44. The method of claim 43 further comprising quantitating the effect of said drug on the metabolism of said first type of cell.

45. The method of claim 38 wherein said first type of cell comprises a cell from heart, stomach, intestine, lung, liver, skin, fat, bone, cartilage, skeletal muscle, smooth muscle, cardiac muscle, bone marrow, muscle, brain, or pancreas.

46. The method of claim 45 wherein said second type of cell comprises malignant or diseased cells.

47. The method of claim 38 wherein said first type of cell comprises liver cells.

48. The method of claim 38 further comprising re-circulating the culture medium.

49. The method of claim 38 wherein the first microscale chamber contains at least one additional type of cell.

50. A method of culturing cells to provide a pharmacokinetic parameter in vitro comparable to that found in vivo comprising:

maintaining a first type of cell in a first microscale chamber, wherein said first microscale chamber is dimensioned to provide a value of at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter found for the same type of cell in vivo, wherein the at least one pharmacokinetic parameter is selected from the group consisting of liquid residence time and liquid-to-cell volume ratio, and wherein the first microscale chamber comprises a first inlet and a first outlet for flow of fluid;

maintaining a second type of cell in a second microscale chamber dimensioned to maintain said second type of cell under conditions that provide a value of at least a second pharmacokinetic parameter in vitro that is comparable to a value of the at least a second pharmacokinetic parameter with respect to said second type of cell in vivo and the second microscale chamber comprises a second inlet and a second outlet for flow of fluid;

flowing fluid through said first and second microscale chambers through at least one microfluidic interconnecting channel; and

culturing said first and said second types of cells in said microscale chambers.

52. The method of claim 50 or 51 further comprising contacting the first type of cell with a material to be tested and evaluating the effect of said material.

53. The method of claims 50 or 51 wherein said first type of cell and said second type of cell are selected from at least one of the group consisting of healthy tissue, diseased tissue, a portion of a tissue biopsy, a portion of skin, a portion of tissue, a portion of an artery, a portion of a vein, a portion of a gastrointestinal tract, a portion of an esophagus, a portion of a colon, a portion of a liver, a portion of an organ, a portion of a heart, a portion of a brain, a portion of a kidney, a portion of a lung, a portion of a muscle, a cell culture, an eukaryotic cell, a plant cell, an animal cell, a mammalian cell, a prokaryotic cell, a primary cell, a tumor cell, a stem cell, a genetically altered cell, a transformed cell, and an immortalized cell.

54. The method of claims 50 or 51 wherein the characteristics of the fluid flow are based on a mathematical model.

55. The method of claim 54 wherein the mathematical model is a physiologically-based pharmacokinetic (“

PBPK”

) model.

51. A method of culturing cells to provide a pharmacokinetic parameter in vitro comparable to that found in vivo comprising:

maintaining a first type of cell in a first microscale chamber, wherein said chamber is dimensioned to provide a value of at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter found for the same type of cell in vivo, wherein the at least one pharmacokinetic parameter is metabolism by cells, wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of fluid, and;

maintaining a second type of cell in a second microscale chamber dimensioned to maintain said second type of cell under conditions that provide a value of at least a second pharmacokinetic parameter in vitro that is comparable to a value for the same at least a second pharmacokinetic parameter with respect to the second type of cell in vivo and the second microscale chamber comprises a second inlet and a second outlet for flow of fluid;

flowing fluid through said first and second microscale chambers through at least one microfluidic interconnecting channel; and

culturing said first type of cell and said second type of cell in said microscale chambers.

56. A method comprising:

forming a first microscale chamber in a device, wherein the first microscale chamber is dimensioned to maintain cells under conditions that provide a value of at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter found in vivo, wherein the at least one pharmacokinetic parameter comprises metabolism by cells, wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio;

forming a second microscale chamber that is dimensioned to maintain cells under conditions that provide a value of at least a second pharmacokinetic parameter in vitro that is comparable to a value for the at least a second pharmacokinetic parameter found in vivo, wherein the at least a second pharmacokinetic parameter comprises metabolism by cells, wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio;

adding, forming, or providing at least one microfluidic interconnecting channel in fluid communication with the first and second microscale chambers, wherein said at least one channel is configured to provide fluid flow through said first and second microscale chambers.

57. The method of claim 56 further comprising adding a debubbler that is configured to remove bubbles from fluid that flows in, through, or in proximity to the microscale chambers.

58. The method of claim 57 wherein the debubbler is located within the device.

59. The method of claim 57 wherein the debubbler is located externally to the device.

60. The method of claim 57 wherein adding, forming or providing for the debubbler further comprises forming, containing, assembling, making, constituting, or inserting the debubbler in or into the device.

61. The method of claim 57 wherein the debubbler is located within a portion of the at least one microfluidic channel.

62. A method of culturing cells to provide a pharmacokinetic parameter in vitro comparable to that found in vivo comprising:

maintaining a first type of cell in a device comprising a first microscale chamber, wherein said first microscale chamber is dimensioned to provide a value of at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter found for the same type of cell in vivo, wherein the pharmacokinetic parameter is selected from the group consisting of liquid residence time and liquid to cell volume ratio, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of culture medium; and

culturing said first type of cell in said first microscale chamber.

63. The method of claim 62, wherein said device further comprises

a second microscale chamber containing a second type of cell, wherein the second microscale chamber comprises a second inlet and a second outlet for flow of culture medium;

and

a microfluidic channel interconnecting the first and second microscale chambers wherein the microfluidic channel is dimensioned to transport culture medium.

64. A method of culturing cells to provide a pharmacokinetic parameter in vitro comparable to that found in vivo comprising:

maintaining a first type of cell in a device comprising a first microscale chamber, wherein said first microscale chamber is dimensioned to provide a value of at least one pharmacokinetic parameter in vitro that is comparable to a value for the same at least one pharmacokinetic parameter found for the same type of cell in vivo, wherein the pharmacokinetic parameter is metabolism by cells, wherein metabolism by cells is achieved by at least any one of tissue size ratio, tissue to blood volume ratio, drug residence time, flow rate, circulatory transit time, liquid residence time, and liquid to cell ratio, wherein the first microscale chamber comprises a first inlet and a first outlet for flow of culture medium; and

culturing said first type of cell in said first microscale chamber.

65. The method of claim 64, wherein said device further comprises

a second microscale chamber containing a second type of cell, wherein the second chamber comprises a second inlet and a second outlet for flow of culture medium;

and

a microfluidic channel interconnecting the first and second microscale chambers wherein the microfluidic channel is dimensioned to transport culture medium.