Patent Application
20210198607
The invention relates generally to microfluidic systems, and more particularly to multicompartment microfluidic bioreactors that can be readily assembled, disassembled, interconnected, and isolated and can utilize cylindrical rotary valves, and applications of the same.
The background description provided herein is for the purpose of generally presenting the context of the invention. The subject matter discussed in the background of the invention section should not be assumed to be prior art merely as a result of its mention in the background of the invention section. Similarly, a problem mentioned in the background of the invention section or associated with the subject matter of the background of the invention section should not be assumed to have been previously recognized in the prior art. The subject matter in the background of the invention section merely represents different approaches, which in and of themselves may also be inventions. Work of the presently named inventors, to the extent it is described in the background of the invention section, as well as aspects of the description that may not otherwise qualify as prior art at the time of filing, are neither expressly nor impliedly admitted as prior art against the invention.
There are a variety of means by which a microfluidic bioreactor can be sealed, i.e., assembled or disassembled in a manner that is reversable yet precludes leaks. For example, an upper plate can be compressed against a lower plate by a plurality of screws, as demonstrated by Shah et al. [1] and many others. The clamping force can be applied by a systems of levers and latches, such as disclosed by Brunswig, et al [2]. Alternatively, component layers can be irreversibly bonded to each other, for example as is done with plasma-activated polydimethylsiloxane (PDMS) microfluidic devices. Layers can be held together through autoadhesion, or by a vacuum channel around the periphery of the device. It is possible to create a single-chamber, multiple-layer, fluidic flow cell whose volume in some configurations is adjustable, for example for infra-red transmission spectroscopy (Specac), but these devices do not provide separate delivery and withdrawal fluidic access to more than one fluidic chamber.
In addition, there is also a need to seal the ports of a microfluidic bioreactor to allow connection of the bioreactor to perfusion systems or analytical devices, and also to disconnect them without introducing bubbles or contamination or losing fluid to the external environment. Some microfluidic bioreactors use external Luer-lock shut-off valves, but these valves are bulky and are not suitable for controlled sterilization of the interconnection or the elimination of bubbles. Other bioreactors have a means for controlled connection, but without valving, e.g., disclosed in U.S. Pat. No. 8,533,413 by Brunswig, et al. [2].
Therefore, a heretofore unaddressed need exists in the art to address the aforementioned deficiencies and inadequacies.
In one aspect, the invention relates to a multichamber bioreactor comprising multiple planar layers stacked on each other defining at least one chamber. In one embodiment, each chamber is implemented from a separate fluidic layer, with each fluidic layer having ports and valves independent of the other layers.
In one embodiment, the multichamber bioreactor further comprises a clamping mechanism. The clamping mechanism comprises a housing and retaining means received in the housing and configured to generate a controlled and uniform pressure to secure the stacked multiple planar layers in the housing.
In one embodiment, the stacked multiple planar layers comprise an endothelial microfluidic disc; at least one somatic cell chamber layer disposed on the endothelial microfluidic disc; a microfluidic perfusion disc disposed on the at least one somatic cell chamber layer; a pressure plate disposed on the at least one somatic cell chamber layer; and at least one membrane, each member disposed between the endothelial microfluidic disc and the at least one somatic cell chamber layer, and between two adjacent somatic cell chamber layers when the at least one somatic cell chamber layer has two or more somatic cell chamber layers. The stacked multiple planar layers are placed on a base plate, received in the housing.
In one embodiment, when high shear flow is not required, the stacked multiple planar layers comprise two perfusion discs, but otherwise as described in the paragraph above. The exact layout and depths of the perfusion channels in any of the layers can be adjusted to match the particular needs of the bioreactor.
In one embodiment, each of the pressure plate and the microfluidic perfusion disc has a plurality of through holes defined therein, and aligned to each other in the stacked multiple planar layers, such that a plurality of tubes with flexible lengths is insertable into the through holes of the pressure plate and the microfluidic perfusion disc for connecting to an individual layer.
In one embodiment, the microfluidic perfusion disc has notches formed on its edge and tubing sockets protruded from the microfluidic perfusion disc for connecting a plurality of tubes to an individual layer; the somatic cell chamber layer has tubing sockets protruded from the somatic cell chamber layer for connecting the plurality of tubes to an individual layer; and the pressure plate has notches formed its edge, such that, as assembled, the tubing sockets of the somatic cell chamber layer are received in the tubing sockets of the microfluidic perfusion disc, which are in turn received in the notches of the pressure plate, so as to allow each upper layer to be inserted over a lower layer without having to thread the plurality of tubes through individual holes in each upper layer.
In one embodiment, the multichamber bioreactor further comprises a transwell adapter for accommodating the at least one somatic cell chamber layer to allow culture of cells independent of the bioreactor prior to insertion of, or after extraction of the at least one somatic cell chamber layer.
In one embodiment, the housing has a threaded inner surface, wherein the retaining means comprises at least one threaded retaining ring being operably threaded into the housing to secure the stacked multiple planar layers in the housing.
In one embodiment, the housing has slots formed in a wall of the housing, and wherein the retaining means comprises a retaining ring having pins radially protruded from a peripheral side of the retaining ring being operably fitted into the slots of the housing 401 to secure the stacked multiple planar layers in the housing. In one embodiment, the slots are L-shape slots.
In one embodiment, the retaining means comprises an expanding clamp pressing outwards against an inner surface of the housing to be held in place by force of friction between the sides of the expanding clamp and the inner surface of the housing.
In one embodiment, the housing includes an internally and externally threaded, notched crown housing, and wherein the retaining means comprises an externally-threaded retaining ring inside the crown; two internally threaded retaining rings outside the crown; a fluidic interface bottom support; a fluidic interface top support; and a microfluidic interface disposed between the fluidic interface bottom support and the fluidic interface top support, wherein the fluidic interface bottom and top supports are designed to provide mechanical support for insertion of tubes or ribbon connectors into the fluidic and to protect the fluidic during handling of the multichamber bioreactor.
In one embodiment, the microfluidic interface comprises conduits embossed on underside and vertical ports. In one embodiment, the two ports and channels furthest from an axis of the conduits are connected to an endothelial chamber of the endothelial microfluidic disc, and the four ports and channels closest to the axis are connected to a stromal cell chamber of the somatic cell chamber layer. In one embodiment, the ports and channels for one layer are directed towards one slot of the crown housing, and the ports for the other layer are directed to a different slot of the crown housing.
In one embodiment, the microfluidic interface further comprises shut-off valves coupled between the conduits and the ports.
In one embodiment, the multichamber bioreactor is operably connected to a perfusion controller, and/or a second, downstream multichamber bioreactor, by at least one connector assembly.
In one embodiment, the multichamber bioreactor further comprises valves integrated into input and output fluidic lines to enable sterilization and washing of open ports of the fluidic connections and elimination of bubbles during the connection process before the ports are connected to the bioreactor chambers and external support equipment, and eliminate leakage of fluid when disconnected.
In one embodiment, the multichamber bioreactor is insertable into a holder whose size is consistent with a standard well plate and whose individual chambers can be cultured separately prior to assembly, with a rotating or friction-fit retaining ring or rings within a structure with a hollow cylindrical region that aligns and holds components together without need for irreversible bonding or multiple fasteners or levers.
In one embodiment, the multichamber bioreactor is rapidly disassembled for imaging or analysis of cellular contents or for their interfacing to other systems and components. In one embodiment, the multichamber bioreactor is operable with an arbitrary number of chambers in a vertical stack without modifications to hardware.
In another aspect, the invention relates to a clamping device, comprising a housing; and retaining means received in the housing and configured to generate a controlled and uniform pressure to secure a layered, planar multi-chamber microfluidic bioreactor in the housing.
In one embodiment, the housing has a threaded inner surface, wherein the retaining means comprises at least one threaded retaining ring being operably threaded into the housing to secure the layered, planar multi-chamber microfluidic bioreactor in the housing.
In one embodiment, the housing has slots formed in a wall of the housing, and wherein the retaining means comprises a retaining ring having pins radially protruded from a peripheral side of the retaining ring being operably fitted into the slots of the housing to secure the layered, planar multi-chamber microfluidic bioreactor in the housing. In one embodiment, the slots are L-shape slots.
In one embodiment, the retaining means comprises an expanding clamp pressing outwards against an inner surface of the housing to be held in place by force of friction between the sides of the expanding clamp and the inner surface of the housing.
In one embodiment, the housing includes an internally and externally threaded, notched crown housing, and wherein the retaining means comprises an externally-threaded retaining ring, and two internally threaded retaining rings.
In yet another aspect, the invention relates to a rotary cylindrical valve, comprising a valve cam; and a valve shaft coupled with the valve cam for rotating the valve cam.
In one embodiment, the rotary cylindrical valve further comprises a valve selection lever affixed to the valve cam for positioning grooves of the valve cam over valve-actuating balls that are secured over microfluidic conduits.
In one embodiment, different angular positions of the valve selection lever permit different combinations of the microfluidic conduits to be opened or closed, based on pressure applied to the valve-actuating balls or cylindrical pins.
In one embodiment, the valve cam comprises a configurable, modular valve cam including different combinations of one-lobe cams and two-lobe cams with their relative angular alignments being set by protrusions that mate with sockets on a face of an adjacent cam.
In one embodiment, the rotary cylindrical valve is actuated by a rotating cylinder that has either detents or protrusions that act upon a ball or a rod or pin to close a desired fluidic channel.
In one embodiment, the rotary cylindrical valve is integrally cast or injection molded as part of a microfluidic system so as to interface with a valve actuator without requiring additional clamping components.
In one embodiment, the valve cam comprises a spindle cam, wherein in operation, valve-actuating ball flow restrictors or actuating pins or rods compress a channel membrane to close microfluidic channels under the actuating elements.
In one embodiment, the spindle cam is adapted to allow activation or deactivation of single or multiple channels in a specific, preprogrammed sequence.
In a further aspect, the invention relates to a rotating cylindrical valve comprising actuators having elevated actuating surfaces for providing pumping functions by sequential compression of a longitudinal fluidic conduit.
In one embodiment, the conduit is either opened or closed depending upon an angular position of the actuators.
In one embodiment, the actuators have a combination of raised and/or recessed regions to enable sequential opening and closing of valves as so to perform different valving and pumping functions.
In one embodiment, the actuators have a variety of widths to control the number of channels being opened or closed.
In one aspect, the invention relates to a valve comprising a valve body being integral to a microfluidic interconnect, the valve body defining a cavity; and a rotary cylindrical valve actuator operably rotating in the cavity.
In one embodiment, within the cavity, there are raised regions beneath which fluidic conduits are located, the raised regions being operably compressed by rotating the rotary cylindrical valve actuator.
In one embodiment, within the cavity, planar surfaces at a top of the cavity maintain an alignment of the rotary cylindrical valve actuator as it rotates and delivers compressive forces to the conduits underneath.
In another aspect, the invention relates to a microfluidic interconnect system comprising a valve body defining a cavity in which a rotary cylindrical valve actuator operates.
In one embodiment, within the cavity, there are raised regions beneath which fluidic conduits are located, the raised regions being operably compressed by regions of the rotary valve actuator without grooves.
In one embodiment, within the cavity, planar surfaces at a top of the cavity maintain an alignment of the rotary cylindrical valve actuator as it rotates and delivers compressive forces to the conduits underneath.
In one embodiment, the conduits connect ports to internal conduits that carry fluids to the connected microfluidic bioreactor, perfusion controller, or other instrument.
In one embodiment, the conduits are formed in either a lower surface of the valve body or an upper surface of a membrane that seals the conduits.
These and other aspects of the invention will become apparent from the following description of the preferred embodiment taken in conjunction with the following drawings, although variations and modifications therein may be affected without departing from the spirit and scope of the novel concepts of the disclosure.
The accompanying drawings illustrate one or more embodiments of the invention and, together with the written description, serve to explain the principles of the invention. Wherever possible, the same reference numbers are used throughout the drawings to refer to the same or like elements of an embodiment.
The invention will now be described more fully hereinafter with reference to the accompanying drawings, in which exemplary embodiments of the invention are shown. The invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Like reference numerals refer to like elements throughout.
The terms used in this specification generally have their ordinary meanings in the art, within the context of the invention, and in the specific context where each term is used. Certain terms that are used to describe the invention are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the invention. For convenience, certain terms may be highlighted, for example using italics and/or quotation marks. The use of highlighting and/or capital letters has no influence on the scope and meaning of a term; the scope and meaning of a term are the same, in the same context, whether or not it is highlighted and/or in capital letters. It will be appreciated that the same thing can be said in more than one way. Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein, nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification, including examples of any terms discussed herein, is illustrative only and in no way limits the scope and meaning of the invention or of any exemplified term. Likewise, the invention is not limited to various embodiments given in this specification.
It will be understood that when an element is referred to as being “on” another element, it can be directly on the other element or intervening elements may be present therebetween. In contrast, when an element is referred to as being “directly on” another element, there are no intervening elements present. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items.
It will be understood that, although the terms first, second, third, etc. may be used herein to describe various elements, components, regions, layers and/or sections, these elements, components, regions, layers and/or sections should not be limited by these terms. These terms are only used to distinguish one element, component, region, layer or section from another element, component, region, layer or section. Thus, a first element, component, region, layer or section discussed below can be termed a second element, component, region, layer or section without departing from the teachings of the invention.
It will be understood that when an element is referred to as being “on,” “attached” to, “connected” to, “coupled” with, “contacting,” etc., another element, it can be directly on, attached to, connected to, coupled with or contacting the other element or intervening elements may also be present. In contrast, when an element is referred to as being, for example, “directly on,” “directly attached” to, “directly connected” to, “directly coupled” with or “directly contacting” another element, there are no intervening elements present. It will also be appreciated by those of skill in the art that references to a structure or feature that is disposed “adjacent” to another feature may have portions that overlap or underlie the adjacent feature.
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprises” and/or “comprising,” or “includes” and/or “including” or “has” and/or “having” when used in this specification specify the presence of stated features, regions, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, regions, integers, steps, operations, elements, components, and/or groups thereof.
Furthermore, relative terms, such as “lower” or “bottom” and “upper” or “top,” may be used herein to describe one element's relationship to another element as illustrated in the figures. It will be understood that relative terms are intended to encompass different orientations of the device in addition to the orientation shown in the figures. For example, if the device in one of the figures is turned over, elements described as being on the “lower” side of other elements would then be oriented on the “upper” sides of the other elements. The exemplary term “lower” can, therefore, encompass both an orientation of lower and upper, depending on the particular orientation of the figure. Similarly, if the device in one of the figures is turned over, elements described as “below” or “beneath” other elements would then be oriented “above” the other elements. The exemplary terms “below” or “beneath” can, therefore, encompass both an orientation of above and below.
Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and the present disclosure, and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.
As used herein, “around,” “about,” “substantially” or “approximately” shall generally mean within 20 percent, preferably within 10 percent, and more preferably within 5 percent of a given value or range. Numerical quantities given herein are approximate, meaning that the terms “around,” “about,” “substantially” or “approximately” can be inferred if not expressly stated.
As used herein, the terms “comprise” or “comprising,” “include” or “including,” “carry” or “carrying,” “has/have” or “having,” “contain” or “containing,” “involve” or “involving” and the like are to be understood to be open-ended, i.e., to mean including but not limited to.
As used herein, the phrase “at least one of A, B, and C” should be construed to mean a logical (A or B or C), using a non-exclusive logical OR. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items.
The description below is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses. The broad teachings of the invention can be implemented in a variety of forms. Therefore, while this invention includes particular examples, the true scope of the invention should not be so limited since other modifications will become apparent upon a study of the drawings, the specification, and the following claims. For purposes of clarity, the same reference numbers will be used in the drawings to identify similar elements. It should be understood that one or more steps within a method may be executed in different order (or concurrently) without altering the principles of the invention.
Our U.S. patent application Ser. No. 15/776,524 (the '524 application), filed May 16, 2018, entitled “Multicomponent Layered and Stackable Microfluidic Bioreactors and Applications of Same”, by John P. Wikswo et al., which is incorporated herein by reference in its entirety, discloses a device that specifically meets unaddressed needs in the art: a device that provides shear-flow induced polarization of endothelial cells; a device that presents small, physiologically realistic fluid volumes to cells that do not dilute concentration-based signaling molecule feedback systems to the point of loss of function or detectability; a modular, stackable device that allows the inclusion of more than two cell types, and the ability to use electrical recordings to monitor neural cell activity in situ.
Our another U.S. patent application Ser. No. 16/012,900 (the '900 application), filed Jun. 20, 2018, entitled “Interconnections of Multiple Perfused Engineered Tissue Constructs and Microbioreactors, Multi-microformulators and Applications of Same”, by John P. Wikswo et al., which is incorporated herein by reference in its entirety, discloses a device that specifically meets unaddressed needs in the art: a device that interconnects separate microfluidic modules in a sterile, low volume manner; a device that avoids fluid loss and introduction of air bubbles; a device that allows ready sterilization of the interior and exterior of the fluid interconnect pathways; a device that allows timed variation in the concentration of key drugs, nutrients, or toxins over an extended period of time. The need for these capabilities is reviewed by Wikswo et al, [3]. The need for proper scaling of the microfluidic bioreactor and tubing volumes to the cell volumes is discussed in Wikswo et al. [4]. Neither article presents specific solutions to these needs.
Our previous invention of the normally closed rotary planar valve [5] presents a valve concept that can be readily implemented in microfluidics, and our '900 application presents a toggle valve suitable for operating a simple means to sterilize, wash, and connect two bioreactor modules. Neither of these inventions is optimized for the sterilization, wash, and connect operations that are needed in the invention described in the current patent application.
Objectives of this invention are, among other things, to refine, extend and integrate the devices disclosed in our '524 application and '900 application.
In certain aspects of the invention, the newly invented device incorporates elements disclosed in the '524 application and the '900 application, and primarily addresses shortcomings that previously made the two earlier technologies difficult to combine into a functional whole. The device does not incorporate all the functions of the two earlier devices, but embodies the functions from each that are most valuable when combined and integrated in a manner that was not obvious at the time of the '524 application and the '900 application. In some embodiments, the device also provides an improvement to a function of one of the devices disclosed in the '524 application and the '900 application.
This system of nested cup-like inserts is limited in several ways. First, with the current design, the addition of layers changes the footprint of the system as a whole. This makes standardization more difficult, and interface of the stacked insert bioreactor with other components, like interconnect valves, more complicated. Because the size difference between inner and outer cup defines the number of possible layers in a given configuration, more pair combinations are necessary to ensure flexibility in device application, and more parts are potentially left unused. Second, because integrity of the mated surfaces of the nested inserts is critical for proper sealing and operation of the device, the magnitude and uniformity of the clamping force required to hold the nested inserts together is also critical. However, no clamp is provided as part of the device, and one must assume some unspecified means to hold the stack together. Finally, the cup-like shape of the inserts present a large, multi-surface area that must remain sealed for proper operation, complicating the design and fabrication of the injection molds to create each of the different-sized nested layers. In addition, this design does not ensure that the vertical clamping that holds the inserts together will be uniformly applied to the tangential surfaces that must be sealed to prevent leaks between the connections of the rim of each insert and the chambers created at the bottom by the vertical space between two adjacent inserts.
The embodiments present in the '900 application are limited to the control of microbioreactors integral to a Perfusion Controller module and management of interconnections between modules. Nothing is presented either about how separate chambers within a microfluidic bioreactor can be sealed together or connected or disconnected from their associate perfusion controller pumps and valves.
In one aspect, the invention relates to a multichamber bioreactor comprising multiple planar layers stacked on each other defining at least one chamber, and secured by a clamping mechanism.
In one embodiment, each chamber is implemented from a separate fluidic layer, with each fluidic layer having ports and valves independent of the other layers.
In one embodiment, the clamping mechanism includes a housing and retaining means received in the housing and configured to generate a controlled and uniform pressure to secure the stacked multiple planar layers in the housing.
In one embodiment, the stacked multiple planar layers comprise an endothelial microfluidic disc; at least one somatic cell chamber layer disposed on the endothelial microfluidic disc; a microfluidic perfusion disc disposed on the at least one somatic cell chamber layer; a pressure plate disposed on the at least one somatic cell chamber layer; and at least one membrane, each member disposed between the endothelial microfluidic disc and the at least one somatic cell chamber layer, and between two adjacent somatic cell chamber layers when the at least one somatic cell chamber layer has two or more somatic cell chamber layers. The stacked multiple planar layers are placed on a base plate, received in the housing.
In one embodiment, the stacked multiple planar layers comprise microfluidic perfusion discs disposed on each side of the at least one somatic cell chamber layer; a pressure plate disposed on the at least one somatic cell chamber layer; and the at least one membrane, each member disposed between the endothelial microfluidic disc and the at least one somatic cell chamber layer, and between two adjacent somatic cell chamber layers when at the least one somatic cell chamber has two or more somatic cell chamber layers. The stacked multiple planar layers are placed on a base plate, received in the housing. The exact topology and height of the perfusion channels in the microfluidic perfusion or endothelial disks can be adjusted as required by the specific cells and tissues to be cultured in the bioreactor.
In one embodiment, each of the pressure plate and the microfluidic perfusion disc has a plurality of through holes defined therein, and aligned to each other in the stacked multiple planar layers, such that a plurality of tubes with flexible lengths is insertable into the through holes of the pressure plate and the microfluidic perfusion disc for connecting to an individual layer.
In one embodiment, the microfluidic perfusion disc has notches formed on its edge and tubing sockets protruded from the microfluidic perfusion disc for connecting a plurality of tubes to an individual layer; the somatic cell chamber layer has tubing sockets protruded from the somatic cell chamber layer for connecting the plurality of tubes to an individual layer; and the pressure plate has notches formed its edge, such that, as assembled, the tubing sockets of the somatic cell chamber layer are received in the tubing sockets of the microfluidic perfusion disc, which are in turn received in the notches of the pressure plate, so as to allow each upper layer to be inserted over a lower layer without having to thread the plurality of tubes through individual holes in each upper layer.
In one embodiment, the multichamber bioreactor further includes a transwell adapter for accommodating the at least one somatic cell chamber layer to allow culture of cells independent of the bioreactor prior to insertion of, or after extraction of the at least one somatic cell chamber layer.
In one embodiment, the housing has a threaded inner surface, wherein the retaining means includes at least one threaded retaining ring being operably threaded into the housing to secure the stacked multiple planar layers in the housing.
In one embodiment, the bioreactor housing has a removable lower retaining ring rather than a retaining lip that is part of the housing. In either case, a separate retaining ring to compress the bioreactor layers.
In one embodiment, the housing has slots formed in a wall of the housing, and wherein the retaining means includes a retaining ring having pins radially protruded from a peripheral side of the retaining ring being operably fitted into the slots of the housing 401 to secure the stacked multiple planar layers in the housing. In one embodiment, the slots are L-shape slots.
In one embodiment, the retaining means includes an expanding clamp pressing outwards against an inner surface of the housing to be held in place by force of friction between the sides of the expanding clamp and the inner surface of the housing.
In one embodiment, the housing includes an internally and externally threaded, notched crown housing, and wherein the retaining means includes an externally-threaded retaining ring;
two internally threaded retaining rings; a fluidic interface bottom support; a fluidic interface top support; and a microfluidic interface disposed between the fluidic interface bottom support and the fluidic interface top support, wherein the fluidic interface bottom and top supports are designed to provide mechanical support for insertion of tubes or ribbon connectors into fluidic and to protect the fluidic during handling of the multichamber bioreactor.
In one embodiment, the microfluidic interface includes conduits embossed on underside and vertical ports. In one embodiment, the two ports and channels furthest from an axis of the conduits are connected to an endothelial chamber of the endothelial microfluidic disc, and the four ports and channels closest to the axis are connected to a stromal cell chamber of the somatic cell chamber layer. In one embodiment, the ports and channels for one layer are directed towards one slot of the crown housing, and the ports for the other layer are directed to a different slot of the crown housing.
In one embodiment, the microfluidic interface further includes shut-off valves coupled between the conduits and the ports.
In one embodiment, the multichamber bioreactor is operably connected to a perfusion controller, and/or a second, downstream multichamber bioreactor, by at least one connector assembly.
In one embodiment, the multichamber bioreactor further includes valves integrated into input and output fluidic lines to enable sterilization and washing of open ports of the fluidic connections and elimination of bubbles during the connection process before the ports are connected to the bioreactor chambers and external support equipment, and eliminate leakage of fluid when disconnected.
In one embodiment, the multichamber bioreactor is insertable into a holder whose size is consistent with a standard well plate and whose individual chambers can be cultured separately prior to assembly, with a rotating or friction-fit retaining ring or rings within a structure with a hollow cylindrical region that aligns and holds components together without need for irreversible bonding or multiple fasteners or levers.
In one embodiment, the multichamber bioreactor is capable of being rapidly dis-assembled for analysis of cellular contents or for their interfacing to other systems and components.
In one embodiment, the multichamber bioreactor is operable with an arbitrary number of chambers in a vertical stack without modifications to hardware.
In yet another aspect, the invention relates to a rotary cylindrical valve, comprising a valve cam; and a valve shaft coupled with the valve cam for rotating the valve cam.
In one embodiment, the rotary cylindrical valve further includes a valve selection lever affixed to the valve cam for positioning grooves of the valve cam over valve-actuating balls that are secured over microfluidic conduits.
In one embodiment, different angular positions of the valve selection lever permit different combinations of the microfluidic conduits to be opened or closed, based on pressure applied to the valve-actuating balls or cylindrical pins.
In one embodiment, the valve cam includes a configurable, modular valve cam including different combinations of one-lobe cams and two-lobe cams with their relative angular alignments being set by cell chamber layer 283 with sockets on a face of an adjacent cam.
In one embodiment, the rotary cylindrical valve is actuated by a rotating cylinder that has either detents or protrusions that act upon a ball or a rod or pin to close a desired fluidic channel.
In one embodiment, the rotary cylindrical valve is integrally cast or injection molded as part of a microfluidic system so as to interface with a valve actuator without requiring additional clamping components.
In one embodiment, the valve cam includes a spindle cam, wherein in operation, valve-actuating ball flow restrictors or actuating pins or rods compress a channel membrane to close microfluidic channels under the valve-actuating balls.
In one embodiment, the spindle cam is adapted to allow activation or deactivation of single or multiple channels in a specific, preprogrammed sequence.
In a further aspect, the invention relates to a rotating cylindrical valve comprising actuators having elevated actuating surfaces for providing pumping functions by sequential compression of a longitudinal fluidic conduit.
In one embodiment, the conduit is either opened or closed depending upon an angular position of the actuators.
In one embodiment, the actuators have a combination of raised and/or recessed regions to enable sequential opening and closing of valves as so to perform different valving and pumping functions.
In one embodiment, the actuators have a variety of widths to control the number of channels being opened or closed.
In one aspect, the invention relates to a valve comprising a valve body being integral to a microfluidic interconnect, the valve body defining a cavity; and a rotary cylindrical valve actuator operably rotating in the cavity.
In one embodiment, within the cavity, there are raised regions beneath which fluidic conduits are located, the raised regions being operably compressed by rotating the rotary cylindrical valve actuator.
In one embodiment, within the cavity, planar surfaces at a top of the cavity maintain an alignment of the rotary cylindrical valve actuator as it rotates and delivers compressive forces to the conduits underneath.
In another aspect, the invention relates to a microfluidic interconnect system comprising a valve body defining a cavity in which a rotary cylindrical valve actuator operates.
In one embodiment, within the cavity, there are raised regions beneath which fluidic conduits are located, the raised regions being operably compressed by regions of the rotary valve actuator without grooves.
In one embodiment, within the cavity, planar surfaces at a top of the cavity maintain an alignment of the rotary cylindrical valve actuator as it rotates and delivers compressive forces to the conduits underneath.
In one embodiment, the conduits connect ports to internal conduits that carry fluids to the connected microfluidic bioreactor, perfusion controller, or other instrument.
In one embodiment, the conduits are formed in either a lower surface of the valve body or an upper surface of a membrane that seals the conduits.
Without intent to limit the scope of the invention, exemplary embodiments of the invention are given below. For purposes of clarity, the same reference numbers will be used in the drawings to identify similar elements.
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All embodiments up to this point have utilized discrete tubes to access the fluidic networks and chambers in each layer. Since this approach requires that the tubes connected to lower layers need to pass down through the upper layers, it is not possible to combine layers that have already been intubated. The embodiment shown in
A limitation presented by each of these tubing implementations is that the insertion of additional chambers in the bioreactors requires that additional space be allocated for the tubing from lower layers to pass through upper ones. The tubing, guided by either holes or slots, also can limit the area of each layer available for fluidic channels within that layer. An alternative embodiment that addresses these problems involves having the fluidics from each layer exit the housing at that layer, which can be accomplished by cutting vertical slots in the tube as shown in
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The two internally threaded rings 502 on the outside of the crown 501 may not be required depending upon the mechanical strength required of the notched crown housing 501 upon compression of the components within the crown by the externally threaded ring 288.
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As shown in
One embodiment of the single-slot format shown in
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The perspective cutaway view of a transwell adapter 600 shown in
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In some applications of the microfluidic bioreactors, such as on the International Space Station with its microgravity environment, it is not acceptable to have fluid leak from the open ends of fluidic conduits that are open to the atmosphere. In many applications, it is not advisable to have microbial contamination enter the reactor through open ports, whether they are wet or dry, and hence there is a need for a means to sterilize recently made connections before fluid passes through them into an established cell culture chamber. Finally, when connecting a microfluidic bioreactor to its perfusion controller or to another bioreactor, it is imperative to eliminate air bubbles from these now-connected ports before fluid is passed through the conduits, which would thereby force any air bubbles in the fluidic connectors into the bioreactor chambers, where they could strip adherent cells from the sides or membranes of these chambers. In selected embodiments of this invention, we provide a valve mechanism that eliminates all of these problems.
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The foregoing description of the exemplary embodiments of the invention has been presented only for the purposes of illustration and description and is not intended to be exhaustive or to limit the invention to the precise forms disclosed. Many modifications and variations are possible in light of the above teaching.
The embodiments were chosen and described in order to explain the principles of the invention and their practical application so as to enable others skilled in the art to utilize the invention and various embodiments and with various modifications as are suited to the particular use contemplated. Alternative embodiments will become apparent to those skilled in the art to which the invention pertains without departing from its spirit and scope. Accordingly, the scope of the invention is defined by the appended claims rather than the foregoing description and the exemplary embodiments described therein.
This application claims priority to and the benefit of U.S. Provisional Patent Application Ser. No. 62/677,468, filed May 29, 2018. This application is also a continuation-in-part application of U.S. Patent Application Ser. No. 16/012,900, filed Jun. 20, 2018, which is a divisional application of U.S. patent application Ser. No. 15/191,092 (the '092 application), filed Jun. 23, 2016, now U.S. Pat. No. 10,023,832, which claims priority to and the benefit of U.S. Provisional Patent Application Ser. Nos. 62/183,571, 62/193,029, 62/276,047 and 62/295,306, filed Jun. 23, 2015, Jul. 15, 2015, Jan. 7, 2016 and Feb. 15, 2016, respectively. The '092 application is also a continuation-in-part application of U.S. patent spplication Ser. Nos. 13/877,925 (the '925 application), 14/363,074 (the '074 application), 14/646,300 (the '300 application) and 14/651,174 (the '174 application), filed Jul. 16, 2013, Jun. 5, 2014, May 20, 2015 and Jun. 10, 2015, respectively. The '925 application, now abandoned, is a national stage entry of PCT Application Serial No. PCT/US2011/055432, filed Oct. 7, 2011, which claims priority to and the benefit of U.S. Provisional Patent Application Ser. No. 61/390,982, filed Oct. 7, 2010. The '074 application, now U.S. Pat. No. 10,078,075, is a national stage entry of PCT Application Serial No. PCT/US2012/068771, filed Dec. 10, 2012, which claims priority to and the benefit of U.S. Provisional Patent Application Ser. Nos. 61/569,145, 61/697,204 and 61/717,441, filed Dec. 9, 2011, Sep. 5, 2012 and Oct. 23, 2012, respectively. The '300 application, now U.S. Pat. No. 9,874,285, is a national stage entry of PCT Application Serial No. PCT/US2013/071026, filed Nov. 20, 2013, which claims priority to and the benefit of U.S. Provisional Patent Application Ser. Nos. 61/729,149, 61/808,455, and 61/822,081, filed Nov. 21, 2012, Apr. 4, 2013 and May 10, 2013, respectively. The '174 application, now U.S. Pat. No. 9,618,129, is a national stage entry of PCT Application Serial No. PCT/US2013/071324, filed Nov. 21, 2013, which claims priority to and the benefit of U.S. Provisional Patent Application Ser. Nos. 61/808,455 and 61/822,081, filed Apr. 4, 2013 and May 10, 2013, respectively. This application is also a continuation-in-part application of U.S. Patent Application Ser. No. 15/776,524, filed May 16, 2018, now allowed, which is a national stage entry of PCT Application Ser. No. PCT/US2016/063586 (the '586 application), filed Nov. 23, 2016, which claims priority to and the benefit of, U.S. Provisional Patent Application Ser. No. 62/259,327, filed Nov. 24, 2015. The '586 application is also a continuation-in-part application of U.S. patent application Ser. Nos. 13/877,925 (the '925 application), 14/363,074 (the '074 application), 14/646,300 (the '300 application), 14/651,174 (the '174 application) and 91,092 (the '092 application), filed Jul. 16, 2013, Jun. 5, 2014, May 20, 2015, Jun. 10, 2015 and Jun. 23, 2016, respectively. Each of the above-identified applications is incorporated herein by reference in its entirety. Some references, which may include patents, patent applications, and various publications, are cited and discussed in the description of the invention. The citation and/or discussion of such references is provided merely to clarify the description of the invention and is not an admission that any such reference is “prior art” to the invention described herein. All references cited and discussed in this specification are incorporated herein by reference in their entireties and to the same extent as if each reference was individually incorporated by reference. In terms of notation, hereinafter, [n] represents the nth reference cited in the reference list. For example, [3] represents the third reference cited in the reference list, namely, Wikswo et al., Engineering Challenges for Instrumenting and Controlling Integrated Organ-on-Chip Systems. IEEE Trans. Biomed. Eng., 60:682-690, 2013.
The invention was made with government support under Grant Nos. 5UG3TR002097-02 and 1U01TR002383-01 awarded by the National Center for Advancing Translational Sciences (NCATS), Contract No. CBMXCEL-XL1-2-001 awarded by the Defense Threat Reduction Agency (DTRA), and contract 2017-17081500003 awarded by the Intelligence Advanced Research Projects Activity (IARPA). The government has certain rights in the invention.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2019/034285 | 5/29/2019 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 62677468 | May 2018 | US | |
| 62183571 | Jun 2015 | US | |
| 62193029 | Jul 2015 | US | |
| 62276047 | Jan 2016 | US | |
| 62295306 | Feb 2016 | US | |
| 61390982 | Oct 2010 | US | |
| 61569145 | Dec 2011 | US | |
| 61697204 | Sep 2012 | US | |
| 61717441 | Oct 2012 | US | |
| 61729149 | Nov 2012 | US | |
| 61808455 | Apr 2013 | US | |
| 61822081 | May 2013 | US | |
| 61808455 | Apr 2013 | US | |
| 61822081 | May 2013 | US | |
| 62259327 | Nov 2015 | US |
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