ORGANOMIMETIC DEVICES AND METHODS OF USE AND MANUFACTURING THEREOF

• US 20160326477A1
• Filed: 12/19/2014
• Published: 11/10/2016
• Est. Priority Date: 12/20/2013
• Status: Active Application

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation), Sony DADC Austria AG (Sony Group Corp.)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation), Jose Fernandez-Alcon
• Inventors: Wen, Norman, Novak, Richard, Umundum, Thomas, Fernandez-Alcon, Jose, Ingber, Donald E., Hamilton, Geraldine A., Hinojosa, Christopher, Domansky, Karel, Levner, Daniel, Thompson, Guy II, Hajipouran Benam, Kambez, Villenave, Remi, Paris, Alfred, Bauer, Georg
• Application Number:
  US15/105,388
• Publication Number:
  US 20160326477A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 63/081 Manufacturing thereof
• CPC Class Codes: B01D 67/0023 by inducing porosity into non porous precursor membranes
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5027 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 B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 23/26 flexible flexible containers for laboratory use B01L3/505
• CPC Class Codes: C12M 25/02 Membranes; Filters filters or filtration in general B01D24/00-B01D41/00
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• 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 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability

Abstract

An organomimetic device includes a microfluidic device that can be used to culture cells in its microfluidic channels. The organomimetic device can be part of dynamic system that can apply mechanical forces to the cells by modulating the microfluidic device and the flow of fluid through the microfluidic channels. The membrane in the organomimetic device can be modulated mechanically via pneumatic means and/or mechanical means. The organomimetic device can be manufactured by the fabrication of individual components separately, for example, as individual layers that can be subsequently laminated together.

Claims

1. A device for simulating a function of a tissue, comprising:

a first microchannel;

a second microchannel;

a membrane located at an interface region between the first microchannel and the second microchannel, the membrane including a first side facing toward the first microchannel and a second side facing toward the second microchannel, the first side having cells of a first type thereon, the membrane separating the first microchannel from the second microchannel and permitting the migration of at least one of cells, particulates, chemicals, molecules, fluids and gases between the first side and the second side;

a first wall portion coupled to the membrane; and

a second wall portion, the membrane being fastened to the second wall portion such that the membrane is modulated by motion of at least one of the first wall portion and the second wall portion.

2. The device of claim 1, wherein the second side has cells of a second type thereon, and the device includes a central microchannel, the membrane dividing the central microchannel into the first microchannel and the second microchannel.

3. The device of claim 1, wherein the first wall portion comprises an elastomeric material that is adapted to move relative to the body by application of a force.

4. The device of claim 3, wherein the force is induced by a pressure differential.

5. The device of claim 3, wherein the force is applied to at least a portion of the body in a direction transverse to the membrane such that the first wall portion flexes in a direction along a plane.

6. The device of claim 1, further comprising:

a first operating channel separated from the first and second microchannels by the first wall portion such that a first pressure differential applied by the first operating channel causes the membrane to stretch or retract in a first direction along a plane.

7. The device of claim 6, further comprising:

a second operating channel separated from the first and second central microchannels by the second wall portion such that a second pressure differential applied by the second operating channel causes the membrane to stretch or retract in a second direction along the plane.

8. The device of claim 6, further comprising at least one rigid element configured to cause the first wall portion to move toward the first operating channel when a positive pressure is applied into the first operating channel, thereby stretching the membrane in the first direction along the plane.

9. The device of claim 1, wherein the first wall portion includes a pivoted lever, whereby a force applied to the lever causes the membrane to stretch or retract in a first direction along the plane.

10. The device of claim 1, wherein the cells of the first type are adhered to the first side of the membrane.

11. The device of claim 2, wherein the cells of the second type are adhered to the second side of the membrane.

12. The device of claim 1, wherein the membrane is substantially rigid.

13. The device of claim 1, wherein the membrane is at least partially flexible.

14. The device of claim 1, wherein at least a portion of the membrane is treated to enhance adhesion of the cells to the membrane.

15. The device of claim 1, wherein at least one surface of the membrane comprises cells of at least two cell types.

16. An organomimetic device comprising:

a first microchannel;

a second microchannel;

a membrane located at an interface region between the first microchannel and the second microchannel, the membrane including a first side facing toward the first microchannel and a second side facing toward the second microchannel, the first side having cells of a first type thereon, the membrane separating the first microchannel from the second microchannel; and

a first engagement element coupled to the membrane whereby the membrane is modulated in at least a first direction along a plane by motion of the first engagement element.

17. The device of claim 16, wherein the second side has cells of a second type thereon.

18. The device of claim 16, wherein the first engagement element can be releasably engaged by an engagement element modulation device, the engagement element modulation device adapted to modulate the motion of the engagement element.

19. The device of claim 16, wherein the first engagement element includes at least one of a bead, a pin, a block, a clamp, a knob, a hole, or any combination thereof.

20. The device of claim 16, wherein the first direction is perpendicular to a fluid flow through the central channel.

21. The device of claim 16, wherein the first direction is parallel to a fluid flow through one of the microchannels.

22. The device of claim 16, wherein the membrane is coupled to a second engagement element, whereby the membrane is modulated in at least a second direction along the plane by motion of the second engagement element.

23. The device of claim 16, wherein the engagement element modulation device is adapted to modulate the movement of first engagement member by modulating a magnetic field.

24. The device of claim 16, wherein the engagement element modulation device includes a solenoid.

25. The device of claim 16, wherein the engagement element modulation device includes a motor.

26. The device of claim 16, wherein the engagement element modulation device includes a pneumatic cylinder.

27. The device of claim 16, wherein the engagement element modulation device includes a shape memory alloy based actuator, a piezo based actuator, or a combination thereof.

28. The device of claim 16, wherein the membrane is substantially rigid.

29. The device of claim 16, wherein the membrane is at least partially flexible.

30. The device of claim 16, wherein at least a portion of the membrane is treated to enhance adhesion of the cells to the membrane.

31. The device of claim 16, wherein at least one surface of the membrane comprises cells of at least two cell types.

32. The device of claim 31, wherein the cells are selected to create an in vitro model that mimics cell behavior of at least a portion of a tissue.

33. The device of claim 31, wherein the cells display at least one characteristic corresponding to a pre-determined physiological endpoint.

34. The device of claim 33, wherein the pre-determined physiological endpoint is selected from the group consisting of a mature state, a differentiated state, a precursor state, a stratified state, a pseudo-stratified state, a confluency state, an inflamed state, an infected state, a stimulated state, an activated state, an inhibitory state, a normal healthy state, a pre-disease state, a disease-specific state, a growth state, a migratory state, a metamorphosing state, or any combinations thereof.

35. The device of claim 16, wherein at least one of the body and the membrane comprises a biocompatible polymer.

36. The device of claim 35, wherein the biocompatible polymer comprises polydimethylsiloxane (PDMS), polymethylmethacrylate (PMMA), polyurethane, styrene-ethylene-butylene-styrene (SEBS), polypropylene, polycarbonate, polyester, cyclic polyolefins, cyclic polyolefin copolymers, or any combinations thereof.

37. The device of claim 16, wherein at least one of the body and the membrane comprises an extracellular matrix polymer, gel, and/or scaffold.

38. The device of claim 16, wherein the body further comprises a second central channel therein;

and a second membrane positioned within the second central channel and extending along a second plane, wherein the second membrane is configured to separate the second central channel to form a third central microchannel and a fourth central microchannel.

39. The device of claim 16, wherein at least one of the first and second microchannels is adapted to fluidically connect to at least one fluid flow-modulation device, the at least one fluid flow modulation device adapted to modulate flow of a liquid or a gas through the first or second microchannel.

40. The device of claim 16, further comprises a cartridge configured to incorporate the device therein.

41. The device of claim 40, wherein the cartridge is configured to establish at least one fluidic connection during operation and optionally provide a sealing of the fluidic connection when not in use.

42. The device of claim 16, wherein the cells of the first type are adhered to the first side of the membrane.

43. The device of claim 17, wherein the cells of the second type are adhered to the second side of the membrane.

61. A method of using an organomimetic device comprising:

a. providing the device of claim 16;

b. introducing a first fluid into the first microchannel of the at least one device; and

c. introducing a second fluid into the second microchannel of the at least one device.

62. The method of claim 61, further comprising mechanically modulating the membrane.

63. The method of claim 61, wherein the mechanical modulation of the membrane causes the membrane to move in at least a first direction along a plane within the channel of the at least one device.

64. The method of claim 61, wherein the mechanical modulation of the membrane is performed by a pneumatic means, a mechanical means, an electrical means, a magnetic means, or a combination thereof.

65. The method of claim 61, wherein the first fluid and/or the second fluid is intermittently or cyclically flowed through the first microchannel and/or the second microchannel.

66. The method of claim 61, wherein the at least one provided device comprises cells on at least one surface of one of the sides of the membrane.

67. The method of claim 61, further comprising introducing cells into the first microchannel, wherein at least a portion of the cells adhere to the first side of the membrane.

68. The method of claim 67, wherein the organomimetic device is used to display at least one characteristic corresponding to a pre-determined physiological endpoint.

69. The method of claim 68, wherein the pre-determined physiological endpoint is selected from the group consisting of a mature state, a differentiated state, a precursor state, a stratified state, a pseudo-stratified state, a confluency state, an inflamed state, an infected state, a stimulated state, an activated state, an inhibitory state, a normal healthy state, a pre-disease state, a disease-specific state, a growth state, a migratory state, a metamorphosing state, or any combinations thereof.

70. The method of claim 61, further comprising creating within the first microchannel an in vitro model that mimics a tissue-specific condition (e.g., in a normal healthy state or in a disease-specific state).

71. The method of claim 67, wherein the cells on the first side of the membrane are selected to create an in vitro model that mimics cell behavior of at least a portion of a tissue, the cells being disease-specific cells isolated from at least one subject or at least one subject population.

72. The method of claim 67, wherein the cells on the first side of the membrane are selected to create an in vitro model that mimics cell behavior of at least a portion of a tissue, the cells being contacted with a condition-inducing agent that is capable of inducing the cells to acquire at least one characteristic associated with the disease-specific state.

73. The method of claim 67, further comprising contacting the cells on the first side of the membrane with a test agent.

74. The method of claim 73, further comprising measuring response of the device and/or the cells on first side of the membrane to the test agent, the first fluid exiting the first microchannel, the second fluid exiting the second microchannel, or any combinations thereof.

75. The method of claim 73, wherein when the cells are disease-specific, the determination of the effect of the test agent identifies a therapeutic agent for treatment of the disease.

76. The method of claim 73, wherein when the cells are patient-specific, the determination of the effect of the test agent identifies a personalized treatment for a subject.

77. The method of claim 61, further comprising connecting the at least one device to a second device.

78. The method of claim 61, further comprising directing the first fluid from the first microchannel of the at least one device to flow to the first microchannel of the second device.

44. An organomimetic device comprising:

a. a first microchannel height-defining layer having a bottom surface and a first microchannel disposed in the bottom surface;

b. a second microchannel height-defining layer having a top surface and a second microchannel disposed in the top surface; and

c. a membrane layer having a membrane portion, the membrane layer being laminated between the bottom surface of the first microchannel height-defining layer and the top surface of the second microchannel height-defining layer,

wherein a first surface portion of the membrane portion provides a lower boundary of the first microchannel and a second surface portion of the membrane portion provides an upper boundary of the second microchannel; and

wherein at least a portion of the first microchannel is aligned with at least a portion of the second microchannel on an opposite side of the membrane portion.

45. The device of claim 44, wherein at least one of the first microchannel height-defining layer and the second microchannel height-defining layer is produced by a process comprising molding.

46. The device of claim 44, wherein the first microchannel height-defining layer comprises:

a first lamination layer having a first microchannel aperture therein, wherein thickness of the first lamination layer defines the height of the first microchannel; and

a first sealing layer disposed on top of the first lamination layer, wherein the first sealing layer is in contact with the first lamination layer and provides a top closure of the first microchannel aperture, thereby forming the first microchannel.

47. The device of claim 44, wherein a top surface of the first microchannel height-defining layer further comprises a substantially rigid layer.

48. The device of claim 44, wherein at least the membrane layer is constructed to include a central region and two side regions on either side of the central region, wherein the central region includes the portion of the membrane separating the first microchannel from the second microchannel.

49. The device of claim 48, wherein a portion of the central region is separated from the two end regions.

50. The device of claim 44, wherein at least one of the first microchannel height-defining layer, the second microchannel height-defining layer, and the membrane layer comprises a biocompatible polymer.

51. The device of claim 50, wherein the biocompatible polymer comprises polydimethylsiloxane (PDMS), polymethylmethacrylate (PMMA), polyurethane, styrene-ethylene-butylene-styrene (SEBS), polypropylene, polycarbonate, polyester, cyclic polyolefins, cyclic polyolefin copolymers, or any combinations thereof.

52. The device of claim 44, wherein at least one of the first microchannel height-defining layer, the second microchannel height-defining layer, and the membrane layer comprises an extracellular matrix polymer, gel and/or scaffold.

53. An organomimetic device produced by a process comprising:

(i) providing at least one first body having a central channel therein along a first axis; and

wherein the central channel has a first wall portion; and

a membrane positioned within the central channel and extending along a plane, wherein the membrane is configured to separate the central channel to form a first central microchannel and a second central microchannel,

wherein the membrane is fastened to the first wall portion whereby the membrane is modulated by motion of the first wall portion; and

wherein the first wall portion comprises an elastomeric material;

(ii) providing a second body having a housing channel therein;

wherein the housing channel has a height that is substantially the same as or greater than the height of the first body; and

a width that is greater than the width of the first body; and

wherein the second body comprises a rigid material; and

(iii) placing the at least one first body within the housing channel of the second body such that the at least one operating chamber forms adjacent to the first wall portion of the first body along the first axis, thereby forming at least one organomimetic device.

54. The device of claim 53, wherein the membrane is substantially rigid.

55. The device of claim 53, wherein the membrane is at least partially flexible.

56. The device of claim 53, wherein at least a portion of the membrane is treated to enhance adhesion of cells to the membrane.

57. The device of claim 53, wherein at least one surface of the membrane comprises cells of at least two cell types.

58. The device of claim 57, wherein the cells are selected to create an in vitro model that mimics cell behavior of at least a portion of a tissue.

59. The device of claim 57, wherein the cells display at least one characteristic corresponding to a pre-determined physiological endpoint.

60. The device of claim 59, wherein the pre-determined physiological endpoint is selected from the group consisting of a mature state, a differentiated state, a precursor state, a stratified state, a pseudo-stratified state, a confluency state, an inflamed state, an infected state, a stimulated state, an activated state, an inhibitory state, a normal healthy state, a pre-disease state, a disease-specific state, a growth state, a migratory state, a metamorphosing state, or any combinations thereof.

79. A mechanical modulation system for stretch actuation of a microfluidic device, the system comprising:

a mechanical actuation arrangement configured to impart an undulating motion along a single plane defined by a microfluidic device mounted within the mechanical modulation system; and

a plurality of opposing connection elements physically connected to the mechanical actuation system, the plurality of opposing connection elements configured to fasten a first location and an second location of a microfluidic device to the opposing connection elements such that the first location and the second location of the microfluidic device are each fixed to one of the connection elements and such that straining of the microfluidic device during cyclical linear motions of a stretch actuation process is transferred to a portion of the microfluidic device between the first location and the opposing second location

80. The system of claim 79, further comprising a sensor arrangement for identifying strain in the microfluidic device.

81. The system of claim 79, wherein the undulating motion is a cyclical linear motion.

82. The system of claim 79, wherein the first location is a first end of a microfluidic device and the second location is an opposing second end of the microfluidic device.

83. The system of claim 79, wherein the microfluidic device includes a membrane with cells adhered thereto.

84. The system of claim 79, wherein fastening of the first location and second location of the microfluidic device to the opposing connection elements includes a plurality of male pin and female slot mating elements.

85. The system of claim 79, wherein the undulating motion during stretch actuation is generally parallel to a long dimension of the microfluidic device, the undulating motion being controlled by at least one of one or more guide rails operatively connected to one of more of the plurality of opposing connection elements.

86. The system of claim 79, wherein the mechanical actuation arrangement includes a motor coupled to a rotating cam configured to impart movement to at least one drive arm that is operatively connected to at least one of the plurality of connection elements.

87. The system of claim 79, wherein the mechanical actuation arrangement is a fluid-based system including one or more piston shafts connected to at least one of the plurality of opposing connection elements.

88. The system of claim 79, wherein the sensor arrangement includes one or more strain gauges and/or linear encoders mounted between the plurality of opposing connection elements.

89. The system of claim 79, wherein the sensor arrangement includes one or more strain gauges and/or linear encoders mounted along a piston shaft and/or linear rail.

90. The system of claim 79, wherein the sensor arrangement includes a linear encoder, a rotary encoder, an optical positioning detector, and/or any combinations thereof.

91. The system of claim 79, wherein the sensor arrangement includes imaging for calibrating the strain associated with the linear motions imparted to the microfluidic device by the mechanical actuation arrangement.

92. The system of claim 79, wherein the first location and the second location of the microfluidic device are fixed to the opposing connection elements such that entry and exit ports positioned at the first location and second location are not exposed to additional strains caused by stretch actuation of the microfluidic device.

93. The system of claim 79, wherein the sensor arrangement includes an imaging device, a limit switch, a proximity switch, and/or any combinations thereof.

94. The system of claim 79, wherein mechanical actuation arrangement includes an electric motor, a voice coil, a solenoid, a piezo driver, and/or any combinations thereof.

95. The system of claim 79, wherein the microfluidic device includes a plurality of microfluidic devices each having a first location and a second location, each of the first locations of the microfluidic devices being fastened to the one of the plurality of opposing connection elements and each of the second locations of the microfluidic devices being fastened to another one of the plurality of opposing connection elements.

96. A microfluidic system for monitoring a behavior of cells, comprising:

a microfluidic device having at least one microchannel in which the cells are disposed; and

a mechanical actuation device for stretching the microfluidic device, the mechanical actuation system including a plurality of opposing connection elements configured to be fastened to a first location and an opposing second location of a microfluidic device.

97. The system of claim 96, wherein the mechanical actuation device for stretching the microfluidic device along a single plane defined by the microfluidic device.

98. The system of claim 96, further comprising a strain monitoring system for identifying a strain in the microfluidic device in response to the stretching.

99. The system of claim 96, wherein the microfluidic device includes a membrane on which the cells are attached.

100. The system of claim 96, wherein the mechanical actuation device imparts undulating motion, the fastening of the first location and the opposing second location of the microfluidic device providing a fixed connection such that the strain of the microfluidic device during the undulating motions of the stretching is transferred to the portion of the microfluidic device between the first location and the opposing second location.

101. The system of claim 96, further comprising entry and exit ports to the at least one microchannel, wherein the microfluidic device is adapted to substantially isolate the entry and exit ports from strains created during the stretching of the microfluidic device.

102. A method of stretch actuation using a mechanical modulation system for a microfluidic device including at least one microchannel in which cells are disposed, the method comprising:

mounting a first location and an second location of the microfluidic device to a first connection element and an second connection element of the mechanical modulation system; and

stretching the microfluidic device, the stretching occurring in response to generally undulating motions imparted to the microfluidic device.

103. The system of claim 102, wherein the stretching of the microfluidic device occurs along a single plane defined by the microfluidic device.

104. The system of claim 102, further comprising identifying strains in the microfluidic device in response to the stretching, the strains being identified by one or more sensor arrangements.

105. The method of claim 102, wherein the microfluidic device includes a membrane on which the cells are disposed.

106. The method of claim 102, wherein the mounting of the first location and the second location of the microfluidic device provides a fixed connection such that strains in the microfluidic device in response to the stretching are transferred to the portion of the microfluidic device between the first location and the second location.