Automated bioculture and bioculture experiments system

• US 20050186671A1
• Filed: 02/24/2005
• Published: 08/25/2005
• Est. Priority Date: 10/02/2000
• Status: Active Grant

• Current Assignee: Koligo Therapeutics, Inc. (Orgenesis Inc.)
• Original Assignee: Tissue Genesis, Inc.
• Inventors: Cannon, Thomas F., Cohn, Laura K., Quinn, Peter D., Kosnik, Paul
• Granted Patent: US 7,906,323 B2
• Time in Patent Office: Days
• US Class Current: 435/297.200
• CPC Class Codes: C12M 23/42 Integrated assemblies, e.g. cassettes or cartridges
• CPC Class Codes: C12M 23/48 Holding appliances; Racks; Supports holding devices for laboratory apparatus B01L9/00
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 37/00 Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination C12M23/38 takes precedence; filtration in general and filters per se B01D24/00-B01D41/00; autoclaves B01J3/04; treatment of microorganisms with electrical or wave energy C12N13/00
• 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: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

The present invention provides a feedback controlled bioculture platform for use as a precision cell biology research tool and for clinical cell growth and maintenance applications. The system provides individual closed-loop flowpath cartridges, with integrated, aseptic sampling and routing to collection vials or analysis systems. The system can operate in a standard laboratory or other incubator for provision of requisite gas and thermal environment. System cartridges are modular and can be operated independently or under a unified system controlling architecture, and provide for scale-up production of cell and cell products for research and clinical applications. Multiple replicates of the flowpath cartridges allow for individual, yet replicate cell culture growth and multiples of the experiment models that can be varied according to the experiment design, or modulated to desired cell development of cell culture end-points. The integral flowpath cartridge aseptic sampling system provides for dynamic analysis of metabolic products or representative cells from the culture.

Claims

1. A culture apparatus for use within an incubator, said apparatus comprising:

a rack for supporting at least one flowpath assembly cartridge;

at least one media flowpath assembly cartridge, said cartridge including;

a housing;

a pump;

at least one valve adapted to prevent or divert media flow;

a control interface;

a sterile media perfusion flowpath loop removable from said housing without breaching flowpath sterility, said media perfusion loop containing in fluid communication with the pump and the at least one valve;

at least one biochamber;

a gas permeable membrane; and

a media reservoir.

2. The apparatus of claim 1, further comprising an access port between said rack and said at least one flowpath assembly cartridge.

3. The apparatus of claim 1, further comprising an access port between said rack and said at least one biochamber.

4. The apparatus of claim 1, wherein said cartridge further comprises at least one noninvasive sensor.

5. The apparatus of claim 1, further comprising a flow sensor.

6. The apparatus of claim 5, wherein said flow sensor comprises a drip chamber.

7. The apparatus of claim 4, further comprising a flow cell removably positionable within said noninvasive sensor.

8. The apparatus of claim 5, wherein said flow sensor is removably positionable within a noninvasive sensor.

9. The apparatus of claim 1, wherein said a gas permeable membrane comprises an oxygenator.

10. The apparatus of claim 9, wherein said oxygenator permits active mass transport of gas into the flowpath.

11. The apparatus of claim 9, wherein said oxygenator permits diffusion of oxygen from an incubator environment into the flowpath.

12. The apparatus of claim 11, wherein said oxygenator is positioned over a valve manifold.

13. The apparatus of claim 12, wherein said valve manifold comprises one or more rotatable cams with associated lifters.

14. The apparatus of claim 13, wherein said membrane is capable of being deformed by the lifters via rotation of said one or more cams.

15. The apparatus of claim 1, wherein said cartridge further comprises a data interface.

16. The apparatus of claim 15, wherein said rack further comprises a data interface for integration with the data interface of the cartridge.

17. The apparatus of claim 1, wherein said rack further comprises a data interface for integration with an external controller.

18. The apparatus of claim 1, wherein said rack further comprises a video camera positioned to obtain images from said at least one biochamber.

19. The apparatus of claim 18, wherein said video camera is a CCD video camera.

20. The apparatus of claim 4, wherein said noninvasive sensor is selected from the group consisting of a pH sensor, a glucose content sensor, an oxygen sensor and a spectroscopy sensor.

21. The apparatus of claim 4, wherein said pump is regulated by feedback control via data received from said noninvasive sensor.

22. The apparatus of claim 1, further comprising a means for providing mechanical forces through the biochamber to a cell or tissue within said biochamber.

23. The apparatus of claim 22, wherein said biochamber further comprises a membrane capable of deformation, wherein said membrane provides force translation into said biochamber for mechanically stimulating a cell or tissue within said biochamber.

24. The apparatus of claim 23, further comprising a cam connectable to said rack or said cartridge and positioned to interface with the membrane, and wherein said membrane deformation is provided by contact with said cam.

25. The apparatus of claim 21, wherein said pump is a multi-head pump.

26. The apparatus of claim 25, wherein said apparatus is capable of generating a cardiac-signature pumping profile.

27. The apparatus of claim 21, wherein said pump is capable of modulation via computer control.

28. The apparatus of claim 27, wherein said modulation creates a pressure head signature.

29. The apparatus of claim 27, wherein said modulation creates a pulse wave signature.

30. The apparatus of claim 29, wherein said pulse wave signature comprises a predetermined physiologic wave form.

31. The apparatus of claim 4, wherein said valve is regulated by feedback control via data received from said noninvasive sensor.

32. The apparatus of claim 1, further comprising a sampling interface.

33. The apparatus of claim 1, wherein said media perfusion loop further comprises a waste reservoir.

34. The apparatus of claim 32, wherein said sampling interface is in communication with an automated sampling device.

35. The apparatus of claim 1 further comprising an injection interface.

36. The apparatus of claim 1, wherein said biochamber further comprises a flexible substrate and said cartridge further comprises:

an actuator positioned adjacent to the biochamber; and

a lever arm that passes through said flexible substrate, wherein said actuator acts upon said lever arm to interact with a tissue within said biochamber.

37. The apparatus of claim 36, wherein said lever arm has a fulcrum located within the flexible substrate.

38. The apparatus of claim 1, wherein said cartridge further comprises:

a rotatable cam shaft connectable to at least one lifter, said lifter having at least one valve actuator for contacting said valve to prevent or divert media flow.

39. The apparatus of claim 1, further comprising a sampling device having:

a fluidic pump for transporting a routing fluid;

a one way flow valve;

a valve for diverting an aliquot of sample from said perfusion loop to said one way flow valve;

wherein said fluidic pump and one way flow valve are connected for transporting routing fluid; and

a means for transporting said aliquot of sample within said routing fluid from said one way flow valve to a collection device or analysis instrument.

40. The apparatus of claim 39, further comprising a connecting means between said means for transporting the aliquot of sample and said collection device.

41. The apparatus of claim 40, wherein said connecting means further comprises a heat source for sterilizing the connection.

42. The apparatus of claim 39, wherein said one way flow valve is a check valve.

43. The apparatus of claim 39, wherein said routing fluid is air.

44. The apparatus of claim 39, further comprising a means for sterilizing the routing fluid.

92. The apparatus of claim 1, wherein said perfusion flowpath loop further comprises a unique digital identifier.

45. A media flowpath assembly cartridge, comprising:

a housing;

a pump;

at least one valve adapted to prevent or divert media flow;

a control interface;

a sterile media perfusion flowpath loop removably attachable to said housing without breaching flowpath sterility, said media perfusion loop containing in fluid communication with the pump and the at least one valve;

at least one biochamber;

a gas permeable membrane; and

a media reservoir.

46. The cartridge of claim 45, further comprising a power source for stand alone operation.

47. The cartridge of claim 45, further comprising a control interface for stand alone operation.

48. The cartridge of claim 47, wherein said control interface provides for operation through an external computer.

49. The cartridge of claim 47, wherein said control interface provides for operation through a user interface.

50. The cartridge of claim 49, wherein said user interface comprises a control pod.

51. The cartridge of claim 50, wherein said control pod comprises at least one embedded microprocessor.

52. The cartridge of claim 50, wherein said control pod comprises a graphical user interface.

53. The cartridge of claim 45, further comprising a sensor and wherein said pump is regulated by feedback control via data received from said sensor.

54. The cartridge of claim 45, further comprising a sensor and wherein said valve is regulated by feedback control via data received from said sensor.

55. The cartridge of claim 45, wherein said gas permeable membrane permits diffusion from an incubator environment into the flowpath.

56. The cartridge of claim 55, further comprising an automated sampling device having:

a fluidic pump for transporting a routing fluid;

a one way flow valve;

a valve for diverting an aliquot of sample from said perfusion loop to said one way flow valve;

wherein said fluidic pump and one way flow valve are connected for transporting routing fluid to said one way flow valve; and

a means for transporting said aliquot of sample within said routing fluid from said one way flow valve to a collection device or analysis instrument.

57. The cartridge of claim 45, further comprising an injection interface.

58. The cartridge of claim 45, further comprising a noninvasive sensor selected from the group consisting of a pH sensor, a glucose sensor, an oxygen sensor and a spectroscopy sensor.

59. The cartridge of claim 45, wherein said at least one biochamber is convertible for use in static cell culture or in a cell perfusion apparatus and comprises:

a first chamber;

a cover;

a seal rendering said first chamber removably connectable to said cover; and

at least one insert positioned between the first chamber and the cover, thereby forming a second chamber.

60. The cartridge of claim 59, wherein said biochamber further comprises a diffuser.

61. The cartridge of claim 59, wherein said seal further comprises two or more sealing interfaces.

62. The cartridge of claim 61, wherein said biochamber further comprises at least one air gap between said two or more sealing interfaces.

63. The cartridge of claim 61, wherein said two or more sealing interfaces are capable of indicating seating of said interfaces by a color change.

64. The cartridge of claim 45, wherein said housing comprises a cutaway for monitoring the biochamber during operation.

65. The cartridge of claim 64, wherein said monitoring is selected from the group consisting of optical viewing, video monitoring, visualization via a microscope, and visualization via an inverted microscope.

66. The cartridge of claim 45, further comprising a light source.

67-91. -91. (canceled)

93. (canceled)