Double-tapered sloped microstructures with embedded microchannel arrays for tissue barrier formation on organ-on-chip platforms

Abstract

An organ-on-chip model includes a first double-sloped microchannel device having a pair of oppositely disposed sloped surfaces, a plurality of microchannels integrated into each sloped surface of the pair of sloped surfaces, wherein the plurality of microchannels being in fluid communication with one another. The organ-on-chip model further includes a pair of cell culture compartments disposed on opposite sides of the first double-sloped microchannel device. The organ-on-chip model is configured to allow cells to attach on top of the pair of sloped surfaces and enable barrier formation to recapitulate tissue barrier function, and the plurality of microchannels are sized to prevent cells from freely moving between the pair of cell culture compartments during cell loading process, and to allow for molecules to diffuse through and allow active cell migration between the pair of cell culture compartments.

Description

DOUBLE-TAPERED SLOPED MICROSTRUCTURES WITH EMBEDDED MICROCHANNEL ARRAYS FOR TISSUE BARRIER FORMATION ON ORGANON-CHIP PLATFORMSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This patent application claims priority from, and incorporates by reference the entire disclosure of, U.S. Provisional Application No. 63 / 657,111 filed on June 6, 2024.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under 2P42ES027704-06 and 1UH3TR003283 awarded by the National Institutes of Health. The government has certain rights in the invention.TECHNICAL FIELD

[0003] The present disclosure relates generally to an organ-on-chip (OOC) platform and more particularly, but not by way of limitation, to an OOC having double-tapered sloped microstructures with embedded microchannel array for tissue barrier formation.BACKGROUND

[0004] This section provides background information to facilitate a better understanding of the various aspects of the disclosure. It should be understood that the statements in this section of this document are to be read in this light, and not as admissions of prior art.

[0005] Microfabrication technologies enable researchers to develop in vitro cell culture platforms that can better mimic in vivo physical tissues, organs, and systems. These systems are known as microphy siological systems (MPS) or organ-on-chip (OOC) platforms, which mimic tissue arrangements observed in living organs and can replicate critical in vivo biological functions and responses. Research on OOC development has advanced significantly in recent decades, showcasing its pivotal role in various applications such as studies on biological pathways, disease modeling, drug discovery', and toxicity' assessment.

[0006] In particular, recapitulating barrier tissues such as placental barrier or blood-brain barrier through OOC platforms have been of high interest. For example, the placenta acts as a protective barrier for the fetus while facilitating biological communication and nutrientexchange between maternal and fetal sides. Studying these barriers using OOC platforms is now becoming a reality. Several methods have been widely utilized and developed to recapitulating these barrier tissues. These include static Transwell culture, porous membranes embedded between microfluidic channels, gelated extracellular matrix (ECM), or arrays of microchannels between microfluidic cell culture compartments. Despite the widespread use of these platforms for recapitulating different tissue barriers for many different applications (pharmaceutical discoveries, toxicity testing, mechanistic studies), limitations persist, hindering the expansion and handling of devices during experiments.

[0007] There are two main approaches in creating barrier tissues composed of two or more different cell types in vitro; horizontal co-culture and vertical co-culture. Vertical coculture through a porous membrane has the advantage of better and easier tight junction formation and thus better for barrier tissue formation in vitro, but is limited in imaging the two different cell types being cultured on the top and bottom side of a porous membrane. It is also difficult to visualize and monitor any cell migration. In addition, in the case where co-culture of three or more different cell types are desired, stacking multiple layers for such co-culture is near impossible. In contrast, horizontal co-culture configuration allows easy visualization of cells cultured, and can be easily expanded to more than two ty pes of cells being co-cultured by simply adding additional compartments. However, because they have horizontal microchannel array that needs to be blocked by cells forming the tight junction and / or barrier, this typically requires the device to be tilted during cell loading process or some other means to block those horizontal channels. Thus, their barrier forming capabilities are typically lower than that of vertical co-culture configuration.SUMMARY OF THE INVENTION

[0008] This summary is provided to introduce a selection of concepts that are further described below in the Detailed Description. This summary is not intended to identity’ key or essential features of the claimed subject matter, nor is it to be used as an aid in limiting the scope of the claimed subject matter.

[0009] In some aspects, an organ-on-chip model includes a first double-sloped microchannel device having a pair of oppositely disposed sloped surfaces, a plurality’ of microchannels integrated into each sloped surface of the pair of sloped surfaces, wherein the plurality ofmicrochannels being in fluid communication with one another. The organ-on-chip model further includes a pair of cell culture compartments disposed on opposite sides of the first double-sloped microchannel device. The organ-on-chip model is configured to allow cells to attach on top of the pair of sloped surfaces and enable barrier formation to recapitulate tissue barrier function, and the plurality of microchannels are sized to prevent cells from freely moving between the pair of cell culture compartments during cell loading process, and to allow for molecules to diffuse through and allow active cell migration between the pair of cell culture compartments.

[0010] In some aspects, each sloped surface of the pair of sloped surfaces is angled to about thirty degrees.

[0011] In some aspects, the organ-on-chip further model includes a plurality' of cell inlets formed into the organ-on-chip model to allow communication of cells into the pair cell culture compartments.

[0012] In some aspects, the organ-on-chip model further includes a housing of the organ-on- chip model is made of a material selected from the group consisting of PDMS. thermoplastic, silica, and glass.

[0013] In some aspects, the double-sloped microchannel device is made of a biocompatible material.

[0014] In some aspects, the double-sloped microchannel device is configured to form cell culture compartments with a width between about 1 micron to 1 centimeter.

[0015] In some aspects, a height of the double-sloped microchannel device and the cell culture compartments is between about 1 micron to 1 centimeter.

[0016] In some aspects, the organ-on-chip model further includes a second double-sloped microchannel device, wherein the orientation of the second double-sloped microchannel device is one of parallel or perpendicular to first double-sloped microchannel device. In some aspects, the first and second double-sloped microchannel devices are arranged within the organ-on-chip model to form a third cell culture compartment allowing multi-layered tissue barriers to be created.

[0017] In some aspects, the double-sloped microchannel device is coated with one of collagen, Matrigel. fibronectin, elastin, parylene, polyimide, or hydrogel.

[0018] In some aspects, the double-sloped microchannel device is integrated with an active media transportation system by connecting the double-sloped microchannel device to a syringe pump for perfusion of cell stimulant into at least one cell culture compartment.

[0019] In some aspects, the double-sloped microchannel device is integrated with electrodes on each side of the barrier and configured for electro-chemical sensing or TEER measurement.

[0020] In some aspects, the organ-on-chip model further includes a first pair of cell inlets in fluid communication with one cell culture compartment of the pair of cell culture compartments.

[0021] In some aspects, the organ-on-chip model further includes a pair of inlets configured to receive an adhesive to affix the double-sloped microchannel device within the organ-on-chip model.

[0022] In some aspects, a double-sloped microchannel device includes a central portion comprising an internal cavity7, a pair of sloped surfaces disposed on opposite sides of the central portion, and a plurality7of microchannels formed through each sloped surface of the pair of sloped surfaces.

[0023] In some aspects, the double-sloped microchannel device includes at least one opening formed through the central portion that is configured to remove a fluid from w ithin the internal cavity.

[0024] In some aspects, each slope surface of the pair of sloped surfaces is configured w ith an angle of about thirty degrees.

[0025] In some aspects, each slope surface of the pair of sloped surfaces is configured with an angle of between about ten and fifty7degrees.

[0026] In some aspects, the plurality of microchannels are sized to prevent cells from freely moving the double-sloped microchannel device, and to allow for molecules to diffuse through and allow active cell migration through the plurality7of microchannels.

[0027] In some aspects, the double-sloped microchannel device is coated with one of collagen, Matrigel. fibronectin, elastin, parylene, polyimide, or hydrogel.BRIEF DESCRIPTION OF THE DRAWINGS

[0028] A more complete understanding of the subject matter of the present disclosure may be obtained by reference to the following Detailed Description when taken in conjunction with the accompanying Drawings wherein:

[0029] FIGS. 1A-1C are multiple views of a device having a double-tapered structure embedded with an array of microchannels for cell-cell communication through diffusion of biomolecules, where the taped structures support cell grow th and barrier formation while also enabling easy microscopy, according to aspects of the disclosure;

[0030] FIG. 2 is a schematic diagram of an OOC incorporating the device of FIGS. 1A-1C, according to aspects of the disclosure; and

[0031] FIGS. 3A-3C are schematic diagrams illustrating additional OOC configurations incorporating one or more of the devices of FIGS. 1 A-l C.DETAILED DESCRIPTION

[0032] It is to be understood that the following disclosure provides many different embodiments, or examples, for implementing different features of various embodiments. Specific examples of components and arrangements are described below to simplify the disclosure. These are, of course, merely examples and are not intended to be limiting. The section headings used herein are for organizational purposes and are not to be construed as limiting the subject matter described.

[0033] The disclosed invention overcomes the challenges discussed above. FIGS. 1A-1C illustrate a double sloped microchannel device 100, according to aspects of the disclosure. Device 100 may be incorporated, for example, into an OOC. FIG. 1A is a perspective view' of device 100, FIG. IB is a cross-sectional side view of device 100, and FIG. 1C is a perspective cross-sectional view of device 100. In the aspect shown in FIGS. 1A-1C, device 100 has an '‘I” shape when viewed from above with a central portion between two arm portions. Device 100 includes a pair of sloped surfaces 102, 104 through which a plurality7of horizontally arrangedmicrochannels 106, 108. respectively, extend. In some aspects, device 100 comprises only a single sloped surface 102 with microchannels 106. and slope surface 108 is replaced with a closed off wall. The plurality of microchannels 106 connect to an internal cavity 110 (best seen in FIGS. IB and 1C) within the central portion of device 100. Device 100 also includes a plurality of openings 112 formed into the central portion of device 100 that allow developer to be applied / removed from internal cavity 110 as a part of the 3D printing process used to create device 100. A plurality of pillars 114 provide structural support for an upper portion of device 100. In FIG. IB, device 100 is cross-sectioned through one pillar 114, while FIG. 1C is crosssectioned through a portion with no pillar 114 (illustrating that internal cavity 110 extends around pillars 114). Device 100 is configured for cells to be cultured on top of the pair of sloped surfaces 102, 104 and on top of the openings of the plurality of microchannels 106 for easy barrier formation, while also allowing easy visualization / microscopy.

[0034] Device 100 can be fabricated by via a two-photon polymerization (2PP)-based 3D printing technique to create the double-tapered barrier structure with microchannels 106, 108 formed through the pair of sloped surfaces 102, 104 as shown in FIGS. 1 A-1C. In some aspects, device 100 maybe coated with ECM (e.g., collagen, matrigel, fibronectin, elastin, and the like) or polymers (e.g., parylene, polyimide, hydrogel, or the like).

[0035] FIG. 2 is a schematic illustrating an OOC that includes device 100 encapsulated within a microfluidic chamber 200 that serves as a barrier to create two separate cell culture compartments 202, 204. Chamber 200 is a housing that encapsulates device 100, which is disposed therein to define a pair of cell culture compartments 202. 204 therebetween. Chamber 200 may be formed from materials including poly dimethyl siloxane (PDMS), thermoplastic, silica, polymethyl methacry late, or glass. Chamber 200 includes a plurality of cell inlets 206(1 )-206(6), with inlets 206(1 )-202(3) in fluid communication with cell culture compartment 202 and inlets 206(4)-206(6) in fluid communication with cell culture compartment 204. Cell inlets 206(1) and 206(3) are connected to cell culture compartment 202 via channels 207(1) and 207(2), respectively, whereas cell inlet 206(2) is positioned directly over cell culture compartment 202. The configuration of cell inlets 206(1) and 206(3) provides the ability to create a flow through cell culture compartment 202 (e g., by introducing cells / media into 206(1) and removing cells / medias from 206(3)). Similarly, cell inlets 206(4) and 206(6) areconnected to cell culture compartment 204 via channels 207(4) and 207(3), respectively, whereas cell inlet 206(5) is positioned directly over cell culture compartment 204.

[0036] Chamber 200 also includes inlets 208(1) and 208(2) that are positioned to receive an adhesive (e.g., a UV glue). In some aspects, applying an adhesive via inlets 208(1) and 208(2) can help secure device 100 within chamber 200 to ensure that cell culture compartments 202. 204 are fluidly separated from one another via device 100 such that fluid communication between cell culture compartments 202, 204 may only occur via the plurality of microchannels 106, 108. The top and bottom flat region of the I-shaped structure illustrated in FIG. 1 also helps create a tight fluidic seal between the left and right side of the double tapered structure when integrated into chamber 200 to prevent fluidic leak between cell culture compartments 202, 204, so that fluidic communication can only occur via the plurality of microchannels 106, 108.

[0037] FIGS. 3A-3C are schematic diagrams illustrating additional OOC configurations incorporating one or more devices 100. FIG. 3 A illustrates a microfluidic chamber 300 that includes one device 100. The OOC of FIG. 3 A is similar to the design shown in FIG. 2, but the arrangement of the cell inlets has been changed. Cell inlets 306(1) and 306(2) are connected to cell culture compartment 302 via channels, and cell inlets 306(3) and 306(4) are connected to cell culture compartment 304 via channels. Inlets 306(3) and 306(6) serve a similar purpose as inlets 208(1) and 208(2). It will be appreciated that inlets may be added or removed to create additional iterations of chamber 300. For example, additional inlets may be positioned over the cell culture compartments.

[0038] FIG. 3B illustrates a microfluidic chamber 310 that includes two devices 100 that are arranged to create three cell culture compartments 312, 313, and 314. Cell inlets 316(1) and 316(2) connect to cell culture compartment 312 and cell inlets 316(5) and 316(6) connect to cell culture compartment 314. Cell inlets 316(3), 316(4), 316(7), and 316(8) sen e a similar purpose as inlets 208(1) and 208(2). It will be appreciated that inlets may be added or removed to create additional iterations of chamber 310. For example, additional inlets may be positioned over the cell culture compartments 312, 313, 314 (i.e., similar to cell inlets 206(2) 206(5) discussed above). FIG. 3C illustrates a microfluidic chamber 320 that includes two devices 100 arranged generally perpendicular to one another to form cell culture compartments 322, 323, 324. Chamber 320 is otherwise similar to chamber 310, with similar configured cell inlets326(1 )-326(8) as shown. It will be appreciated that inlets may be added or removed to create additional iterations of chamber 320. For example, additional inlets may be positioned over the cell culture compartments 322, 323, 324 (i.e., similar to cell inlets 206(2) 206(5) discussed above).

[0039] Advantages of device 100 include:

[0040] (1) Sloped surfaces 102, 104 with embedded microchannels 106, 108 allow real-time monitoring of the confluency and morphology of cell barrier attached on the tapered structure under bright field microscopy, which is not feasible with conventional method utilizing porous membrane for cell separation or horizontal microchannel array where cells forming barrier are attached to the inlet and outlet parts of the microchannels.

[0041] (2) Sloped surfaces 102, 104 are integrated with microchannels 106, 108 to allow communication of cells between each cell culture compartment (e.g., compartments 202, 204). Size, shape, and density of microchannels 106, 108 can be manipulated easily by changing parameters of the 3D model design to control the degree of molecular communication betw een the two cell culture compartments.

[0042] (3) Device 100 allows co-culture of cells in lateral arrangement, so that the platform can be expanded with multiple tapered structures to accommodate more than two cell culture compartments for more complex tissue barriers (e.g., See FIGS. 3A-3C). Additionally, device 100 enables easy fluorescent microscopy of any cell culture compartment, even with complex tissue models, which is difficult on any other type of reported platforms available in the market.

[0043] (4) Sloped surfaces 102, 104 overcome the challenge of creating barrier tissues in horizontal co-culture configurations.

[0044] The methods discussed herein provide a better OOC platform for recapitulating any barrier tissue, where such structure, functions, physiology, and pathophysiology are of interest. This allows expansion of the model for co-culture of multiple (more than two) cell types to mimic complex tissue barrier and real-time monitoring of cell confluency and morphology with brightfield (“BF”) microscopy.

[0045] In more detail, device 100 is designed with two tapered sidewalls (i.e., sloped surfaces 102, 104) that are sloped in opposite directions to facilitate cell attachment and barrier formation on either or both sides of device 100. In some aspects, sloped surfaces 102, 104 are angled at a 30-degree angle, enabling easy monitoring of attached cells under a microscope. In other aspects, the angle may be varied to be greater than or less than 30 degrees (e.g., between about 10 degrees to 50 degrees). Traditional methods, such as the Transwell insert, enable coculturing of two different cell types by allowing communication through pores on the membrane. However, because of the contrast difference under a microscope, neither the top cell layer nor the reverse side cell layer can be easily observed, which prevents evaluation of cell confluency, morphology’, and location under bright field microscopy. Each of these are important parameters to observe during experiments. The only time cells cultured on both side of a porous membrane, such as a polyethylene terephthalate (PET) membrane, can be properly monitored is through end-point fluorescent staining. This staining can only be done at the end of the experiment, which significantly limits the types and ranges of experiments that can be conducted.

[0046] For platforms utilizing vertical walls integrated with arrays of microchannels or extracellular matrix (ECM) for cell separation, forming cell barriers on vertical chamber walls poses challenges for confirming cell attachment under a microscope. Even with fluorescent staining, confirming cell attachment is difficult. Importantly, this setup is not typically conducive to optimal cell attachment. The double-tapered structure of device 100 addresses these challenges by incorporating sloped surfaces 102, 104 for cell barrier formation on each side of device 100. with cell-specific culture compartments 202, 204 located adjacent to each other. This design allows for real-time monitoring of each cell line without the need to sacrifice devices for fluorescent microscopy at each time point of interest.

[0047] To enable co-culture and cell-cell communicarion, as well as molecular diffusion between the cell culture chambers on each side of the sloped barrier, device 100 incorporates the plurality of microchannels 106. Each microchannel 106 has openings measuring about 10 pm x 10 pm (width x height), smaller than the typical diameter of mammalian cells (15-20 pm), to prevent direct mixing of cells between the cell culture compartments during the cell seeding process. The size, length, and density of these microchannels can be easily adjusted to match the tissue microenvironment by modifying parameters of the 3D model design for two-photon polymerization (2PP) printing. This level of customization also provides the disclosed invention with flexibility in engineering the permeability of the double-tapered barrier by manipulating the design of microchannels, since the size of the microchannel dimensions can control the degree of permeability between the cell culture chambers.

[0048] Compared to platforms utilizing porous membranes, which offer limited options for commercially available membranes with different porosities, device 100 provides a more versatile solution. Furthermore, device 100 allows for the manipulation of barrier permeability without the need for extracellular matrix (ECM) such as Matrigel, which could potentially interfere with cell responses during experiments.

[0049] Device 100 enables the co-culture of different cell lines in a lateral arrangement, unlike platforms that utilize porous membranes for cell separation in a vertical format. Layering cells vertically not only affects their visibility under a microscope, as mentioned previously, but also complicates cell seeding and the microfabrication process, particularly when constructing complex tissue interfaces requiring multiple (more than 2) cell layers.

[0050] As depicted in FIG. 3, the integration of multiple double-tapered barrier structures within a microfluidic chamber allows for the creation of additional cell environments. This setup is advantageous for mimicking systems that require more than two cell types to recapitulate biological responses in humans. Moreover, the distances between barrier structures can be manipulated to match the microenvironment of the barrier interface in humans.

[0051] The feasibility of using the current design as an OOC platform mimicking barrier tissue was tested by modeling the human placental barrier. Immortalized human placental trophoblast cells (PTC) and primary human umbilical vein endothelial cells (pHUVEC) were co-cultured on each side of the double-tapered barrier structure encapsulated within a biocompatible polydimethylsiloxane (PDMS) micro-chamber. Nutrients for cell growth were provided by a media reservoir stacked on top of the cell culture chamber. Results demonstrated that cells loaded into the platform could be easily observed under bright field microscopy before and after attachment. The confluency and morphology of cells were monitored over time without sacrificing a device for fluorescent microscopy. Live / dead cell staining and lactate dehydrogenase (LDH) assays confirmed that both PTC and pHUVEC could maintain good viability (>95% live) on the invented platform after 3 days without media replenishment.Following the experiment, cells could be fixed for endpoint analysis such as scanning electron microscopy and immunofluorescent staining of specific biomarkers.

[0052] Placental barrier formation was further confirmed through a perfusion test using fluorescent-labeled dextrans with different molecular weights (3k. 10k, and 70kDa MW) between the two cell culture compartments with and without cells attached to the tapered barrier. A mixture of dextrans was added from the PTC culture chamber, and fluorescent intensity indicating dextran diffusion across the tapered barrier was measured from the pHUVEC culture compartment. Results demonstrated that the acellular barrier allowed an observable amount of dextran molecules to diffuse within two hours, while the permeability of the barrier with PTC and pHUVEC cells was 30 times less than the acellular barrier. From this result, it can be concluded that the placental barrier could successfully form on the tapered structure, limiting perfusion of molecules across the two different cell layers. Trans-epithelial electrical resistance (TEER) was also measured during the three-day culture of PTC on the device, which demonstrated an increase in TEER over time as the cells attached and formed a barrier on the tapered structure.

[0053] It is important to note that while this invention primarily demonstrates the application of recapitulating the human placental barrier, the platform can also be adapted for other OOC devices where barrier tissue functions are mimicked. These may include the blood-brain barrier, intestinal barrier, and skin barrier, offering opportunities for mechanistic studies on disease models, drug development, and even cosmetic development.

[0054] The described model can accommodate different cell types, numbers of barriers, and designs of microchannels, including variations in size, shape, length, and density. Additionally, it can utilize different fabrication materials and techniques to suit specific requirements.

[0055] Depending on resolution and scale requirements, the tapered barrier can be fabricated using various microfabrication and 3D printing techniques, such as material jetting and stereolithography, commonly employed in bioprinting. The material for the tapered barrier can be substituted with other biocompatible polymers or bioinks, allowing for cell observation under bright field microscopy. The cell culture chamber layer, integrated with the tapered barrier, can be fabricated using materials like PDMS, thermoplastic, silica, polymethyl methacry late, or glass.

[0056] Although various embodiments of the present disclosure have been illustrated in the accompanying Drawings and described in the foregoing Detailed Description, it will be understood that the present disclosure is not limited to the embodiments disclosed herein, but is capable of numerous rearrangements, modifications, and substitutions without departing from the spirit of the disclosure as set forth herein.

[0057] The term “substantially” is defined as largely but not necessarily wholly what is specified, as understood by a person of ordinary' skill in the art. In any disclosed embodiment, the terms “substantially”, “approximately”, “generally”, and “about” may be substituted with “within [a percentage] of’ what is specified, where the percentage includes 0.1. 1, 5, and 10 percent.

[0058] The foregoing outlines features of several embodiments so that those skilled in the art may better understand the aspects of the disclosure. Those skilled in the art should appreciate that they may readily use the disclosure as a basis for designing or modifying other processes and structures for carry ing out the same purposes and / or achieving the same advantages of the embodiments introduced herein. Those skilled in the art should also realize that such equivalent constructions do not depart from the spirit and scope of the disclosure, and that they may make various changes, substitutions, and alterations herein without departing from the spirit and scope of the disclosure. The scope of the invention should be determined only by the language of the claims that follow. The term “comprising” within the claims is intended to mean “including at least” such that the recited listing of elements in a claim are an open group. The terms “a”, “an”, and other singular terms are intended to include the plural forms thereof unless specifically excluded.#

[0059] Conditional language used herein, such as, among others, “can”, “might”, “may”, “e.g.”, and the like, unless specifically stated otherwise, or otherwise understood within the context as used, is generally intended to convey that certain embodiments include, while other embodiments do not include, certain features, elements and / or states. Thus, such conditional language is not generally intended to imply that features, elements and / or states are in any way- required for one or more embodiments or that one or more embodiments necessarily include logic for deciding, with or without author input or prompting, whether these features, elements and / or states are included or are to be performed in any particular embodiment.

[0060] While the above detailed description has shown, described, and pointed out novel features as applied to various embodiments, it will be understood that various omissions, substitutions, and changes in the form and details of the devices or algorithms illustrated can be made without departing from the spirit of the disclosure. As will be recognized, the processes described herein can be embodied within a form that does not provide all of the features and benefits set forth herein, as some features can be used or practiced separately from others. The scope of protection is defined by the appended claims rather than by the foregoing description. All changes which come within the meaning and range of equivalency of the claims are to be embraced within their scope.

[0061] Although various embodiments of the method and apparatus of the present invention have been illustrated in the accompanying Drawings and described in the foregoing Detailed Description, it will be understood that the invention is not limited to the embodiments disclosed, but is capable of numerous rearrangements, modifications and substitutions without departing from the spirit of the invention as set forth herein.#

Claims

CLAIMSWhat is claimed is:

1. An organ-on-chip model comprising: a first double-sloped microchannel device having: a pair of oppositely disposed sloped surfaces; and a plurality of microchannels integrated into each sloped surface of the pair of sloped surfaces, the plurality of microchannels being in fluid communication with one another; a pair of cell culture compartments disposed on opposite sides of the first doublesloped microchannel device, wherein the organ-on-chip model is configured to allow cells to attach on top of the pair of sloped surfaces and enable barrier formation to recapitulate tissue barrier function, and wherein the plurality of microchannels are sized to prevent cells from freely moving between the pair of cell culture compartments during cell loading process, and to allow for molecules to diffuse through and allow active cell migration between the pair of cell culture compartments.

2. The organ-on-chip model of claim 1, wherein each sloped surface of the pair of sloped surfaces is angled to about thirty degrees.

3. The organ-on-chip model of claim 1, further comprising a plurality of cell inlets formed into the organ-on-chip model to allow communication of cells into the pair cell culture compartments.

4. The organ-on-chip model of claim 1, wherein a housing of the organ-on-chip model is made of a material selected from the group consisting of PDMS, thermoplastic, silica, and glass.

5. The organ-on-chip model of claim 1, wherein the double-sloped microchannel device is made of a biocompatible material.

6. The organ-on-chip model of claim 1 , wherein the double-sloped microchannel device is configured to form cell culture compartments with a width between about 1 micron to 1 centimeter.

7. The organ-on-chip model of claim 1, wherein a height of the double-sloped microchannel device and the cell culture compartments is between about 1 micron to 1 centimeter.

8. The organ-on-chip model of claim 1, further comprising a second double-sloped microchannel device, wherein the orientation of the second double-sloped microchannel device is one of parallel or perpendicular to first double-sloped microchannel device.

9. The organ-on-chip model of claim 8, wherein the first and second double-sloped microchannel devices are arranged within the organ-on-chip model to form a third cell culture compartment allowing multi-layered tissue barriers to be created.

10. The organ-on-chip model of claim 1, wherein the double-sloped microchannel device is coated with one of collagen, matrigel, fibronectin, elastin, parylene, polyimide, or hydrogel.

11. The organ-on-chip model of claim 1. wherein the double-sloped microchannel device is integrated with an active media transportation system by connecting the doublesloped microchannel device to a syringe pump for perfusion of cell stimulant into at least one cell culture compartment.

12. The organ-on-chip model of claim 1, wherein the double-sloped microchannel device is integrated with electrodes on each side of the barrier and configured for electrochemical sensing or TEER measurement.

13. The organ-on-chip model of claim 1, further comprising a first pair of cell inlets in fluid communication with one cell culture compartment of the pair of cell culture compartments.

14. The organ-on-chip model of claim 1, further comprising a pair of inlets configured to receive an adhesive to affix the double-sloped microchannel device within the organ-on- chip model.

15. A double-sloped microchannel device comprising: a central portion comprising an internal cavity; a pair of sloped surfaces disposed on opposite sides of the central portion; and a plurality of microchannels formed through each sloped surface of the pair of sloped surfaces.

16. The doubled-sloped microchannel device of claim 15, further comprising at least one opening formed through the central portion that is configured to remove a fluid from within the internal cavity.

17. The double-sloped microchannel device of claim 15, wherein each slope surface of the pair of sloped surfaces is configured with an angle of between about ten and fifty degrees.

18. The doubled-sloped microchannel device of claim 15. wherein the plurality of microchannels are sized to prevent cells from freely moving the double-sloped microchannel device, and to allow for molecules to diffuse through and allow active cell migration through the plurality of microchannels.

19. The double-sloped microchannel device of claim 15, wherein the double-sloped microchannel device is coated with one of collagen, Matrigel, fibronectin, elastin, pary lene, polyimide, or hydrogel.

20. An organ-on-chip model comprising: a sloped microchannel device having: a sloped surface extending from a central portion of the device; and a plurality of microchannels integrated into the sloped surface; a cell culture compartment disposed adjacent to the sloped surface, wherein the organ-on-chip model is configured to allow cells to attach on top of the sloped surface and enable barrier formation to recapitulate tissue barrier function.

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