Human-on-a-chip Operating System

Abstract

Human-on-a-chip plate. The human-on-a-chip plate includes at least one 3D culture biochip, at least one microvalve, at least one microfluidic channel, and at least one inlet or outlet plate port. The outlet port from one chip can be connected to another chip through a valve. Multiple chips can be connected in parallel, series or combinations, and one or more chips in the human-on-a-chip plate can be bypassed. Each fluid connection can be changed independently at any time. At least one valve system connects or separates the various elements of the organ-on-a-chip system. At least one inlet port and one outlet port access or block the microchannel. At least one valve system can allow extraction, modification of the model flow map, and introduction or reduction of at least one fluid chamber.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 293,422, filed December 23, 2021. The contents of the above-mentioned patent applications are incorporated by reference in their entireties into this application. [Background technology]

[0002] Developing a successful drug is expensive and time-consuming. Moreover, many drugs fail in the final stages of the development process, causing large sunk costs. One reason drugs fail in the late stages of development is that pharmaceutical companies typically test drugs on cells cultured in cell culture platforms that do not adequately replicate human tissues. 3D organ-on-chip technologies have emerged to create more similar tissue models that solve the problems of structure and assembly of 3D tissues. However, the problem remains of running an operating system system to nourish, maintain, and grow the tissue while reproducing the mechanics and fluid dynamics of the operating system's advanced 3D model, and to measure and analyze the parameters of interest in real time with the operating system's advanced system.

[0003] Current cell culture experiments often do not accurately mimic physiological conditions, using simple approaches that only reproduce a few aspects rather than the entire system. Several factors are closely related to the development and operation of 3D advanced cell culture models. The operating factors span a wide range of design parameters, including the level of model complexity, the connection mechanisms between the various organ models, the pumping, seeding and extraction mechanisms, as well as the measurement, sensing and imaging mechanisms, as well as analytical and predictive techniques. A comprehensive system encompassing all the latter factors is needed to obtain accurate drug predictions from preclinical trials.

[0004] One of the main persistent problems facing current tissue culture semi-automated operating systems is that they require large volumes of material, tens of milliliters, just for seeding, while only using a few microliters of cells or biological material, limiting the cultivation and modeling of tissues using expensive or rare material, such as biopsies.

[0005] Another important aspect in the operation of complex 3D culture models is analytical readout, which limits high-throughput analysis and real-time data collection. Moreover, in current Organ Chips, it is difficult to access cell and media samples for analysis at different time points. Media is defined as a fluid that contains either therapeutic agents, growth factors, nutrients, chemicals, cells in suspension, and / or other molecules, biomolecules or substances that flow through the channels.

[0006] Typical complex 3D cell culture platforms do not allow for real-time imaging and measurement of biological parameters of individual tissues connected to the human-on-a-chip model while manipulating and flowing media within the biochip. One limitation is that the design does not allow for more than one analytical device to be placed close to the chip due to limited space. For example, to model the metastasis of cancer from one tissue to another through a 3D vasculature, it is not possible to image both tissues simultaneously while pumping fluid through the blood vessels. Similarly, much of the valuable data generated is mostly limited to a few data points rather than continuous measurements.

[0007] One key aspect of typical chip designs and inter-organ connection methods is the inability to set up complex experiments with multiple organs while maintaining accurate sensing and communication. These systems lack the communication and sensing found in the human body, and do not have the ability to set up arbitrary experiments with chip patterns.

[0008] One of the main limitations these systems face when extracting and inserting media is the need for large volumes of fluid to compensate for the volume of the tubes in the system. The use of ultra-thin tubes or microchannels eliminates the need for dilution elements used to move fluids in other systems. However, such low diameter tubes maintain high capillary forces and high surface tension within the tube, allowing small volumes of fluid to be moved further from one location to another. The absence of dilution elements also results in a sample that is highly concentrated in the molecules that need to be detected, resulting in more accurate measurements and results.

[0009] One of the key aspects of cell culture experiments is the ability to accurately measure and read physiologically relevant data. Microscopy, spectroscopy, and FTIR are some of the widely used techniques to perform quantitative measurements of biological materials that indicate cell viability, functionality, and metabolism. Most human-on-chip operating systems currently on the market rely on external measurement devices to perform such assays. These tests require the operator to pause the experiment and transfer it to the measurement device. This results in the loss of valuable observations and data between set time points. Such manipulations increase the number of variables, which can have unexpected effects on the experimental results. There is a dire need to implement multiplexed real-time measurements in running cell culture experiments. For example, implementing an on-board triple measurement system consisting of microscopy, spectroscopy, and ATR-FTIR tightly integrated with the operating system would provide more valuable information and a better representation of the experiment while eliminating variables that may form as a result of rearrangements of the culture. The importance of using multiple measurement techniques to perform various reads simultaneously while connected to the same data reader is that it allows obtaining multiplexed analyses and conclusions that would not be possible otherwise. Summary of the Invention

[0010] The present disclosure provides a new and innovative biochip operating system that serves as a cell culture platform to model and manipulate multiple human organs simultaneously. The operating system includes and operates a human-on-a-chip plate. The human-on-a-chip plate is an array of multiple biochips fluidly connected via tubes that model multiple organs of the human body. Cells can be inserted into the biochips and grown into three-dimensional tissues for use in drug testing. Each biochip can include one or more ultra-thin porous plastic cylindrical tubes. Each tube can be accessed from the inside via microfluidic channels or from the area surrounding the tube.

[0011] The present disclosure provides a means to replicate a ductal organoid microenvironment by growing ductal cells on the inner wall of each duct and growing the surrounding tissue from the outside by seeding the cells and delivering media components through a gel from the other side surrounding the duct. Each biochip can be used to replicate ductal or non-ductal tissues including, but not limited to, pancreas, kidney, liver, breast, brain, lung, blood vessels, prostate, fallopian tubes, testes, and lymphatic vessels. A human-on-a-chip plate is an array of biochips, each of which grows and models a specific tissue and is fluidically connected to other biochips through one or more tubes of each biochip. The tubes of the biochips are separated by a valve system that allows the connection between the different biochips. These biochips can be connected in series, parallel, or combinations. Furthermore, the valve system allows the modification of these connections at any point. The chips can also be connected to an extraction chamber, ATR-FTIR, or waste chamber.

[0012] The present disclosure provides a biochip operating system that is a semi-automated or fully automated device that operates a human-on-a-chip plate to deliver and control the flow of fluids containing cell growth media, chemicals and cells to each of the channels of the biochip. The human operating system also includes sensing and measurement devices to measure and control various biological parameters of the tissues grown within the biochip. The actuation of fluid valves and pumps as well as sensors and images are fed to computer software that automates the cell culture and measurement process. The biochip operating system can be used by researchers, pharmaceutical companies and clinicians, including but not limited to, to test drugs or other compounds on complete human models made with cells from biopsies, primary cells, stem cells and / or cell lines. The system can perform, control and measure complete cell culture experiments for a wide range of applications.

[0013] The present disclosure provides a system including devices and methods for delivering and controlling very small (on the order of 0.1 pl) fluid flows containing cells, media, therapeutics, chemicals, reagents and other biological components to individual channels within individual biochips, enabling accurate, repeatable and reproducible biological experiments to be performed on highly similar tissue models.

[0014] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, an operating system comprises at least one biological system-on-a-chip plate, at least one valve means, at least one actuation means for actuating the valve means, at least one fluid control means, at least one analytical device, and a control system for controlling the pump means, the actuation means and the valve means and the analytical device.

[0015] In another aspect of the present disclosure, which may be used in combination with other aspects or combinations of aspects enumerated herein, the analytical device may inspect the biochip using at least one of microscopic images or spectral measurements.

[0016] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the analytical device may inspect the medium within the fluid control means or fluid path.

[0017] In another aspect of the present disclosure, which may be used in combination with other aspects or combinations of aspects enumerated herein, the operating system facilitates, semi-automates, or automates at least one aspect of experimental setup, including at least one of: cleaning, sterilizing, or preparing the system; priming the system with a biocompatible fluid; placing the correct cell type on the chip in the correct location required for the desired experiment; selectively connecting the chip in the correct configuration for the desired experiment; and / or metering the correct amount of media or other compounds into the system.

[0018] In another aspect of the present disclosure, which may be used in combination with other aspects or combinations of aspects enumerated herein, the operating system facilitates, semi-automates, or automates at least one aspect of experimental execution, including at least one of circulating media between chips as required by the experiment, controlling flow rates of fluid movement, possibly extracting samples at specific time points or time intervals, testing using analytical means at specific time points or time intervals, and / or isolating and immobilizing biochips for subsequent analysis.

[0019] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the operating system further comprises a motion stage for moving the biological system-on-chip plate relative to the analytical device and / or the drive means.

[0020] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, a biochip for culturing a multi-vascular tissue comprises a housing, a plurality of interior regions, and a plurality of fluidic media, the housing including the plurality of interior regions, at least one of the interior regions being in permeable or semi-permeable communication with at least one other interior region.

[0021] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, the plurality of interior regions are tubes.

[0022] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least two tubes are arranged with an inlet and an outlet to allow the passage of a first fluid medium.

[0023] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least two tubes are arranged with an inlet and an outlet that are distinct from the first tube to allow for the passage of a second fluid medium that is separate from the first fluid medium.

[0024] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least one region is an interstitial compartment that is external to the tube but internal to the biochip housing.

[0025] In another aspect of the disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the multiple fluid media are the same media.

[0026] In another aspect of the disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the multiple fluid media are different media.

[0027] In another aspect of the disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the plurality of fluid media is a combination of fluid media types.

[0028] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, a biochip fluid control comprises at least one human-on-a-chip including at least one organ-on-a-chip system including at least one tube scaffold interacting with at least one surrounding compartment, at least one valve system controlling flow to each compartment, at least one micropump mechanism capable of pumping and controlling flow to at least one compartment, and at least one actuator controlling valves on the chip, connected to the chip, and controlling fluid pumps to the chip compartments. The at least one valve system can open, close and change the flow paths to all compartments.

[0029] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the at least one valve mechanism may be pneumatically, mechanically, electrically, and / or fluidically actuated.

[0030] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism connects at least one biochip to an operating system.

[0031] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least one valve mechanism connects at least two biochips in a human-on-a-chip connection plate.

[0032] In another aspect of the disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least one valve mechanism controls at least two biochips in a human-on-a-chip connection plate.

[0033] In another aspect of the disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism controls at least one inlet and outlet biochip.

[0034] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism connects and controls the connection of at least one tube to another tube.

[0035] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism connects at least one interstitium to another interstitium.

[0036] In another aspect of the present disclosure, which may be used in combination with other aspects or combinations of aspects recited herein, at least one valve mechanism connects and controls the connection of at least one tube to at least one interstitium.

[0037] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least one valve mechanism connects at least one biochip to at least one measurement device.

[0038] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, the at least one valve mechanism includes at least one bi-stable valve.

[0039] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, the at least one valve mechanism includes at least one on-off valve.

[0040] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, the at least one valve mechanism includes at least one bistable valve and an on-off valve.

[0041] In another aspect of the present disclosure, which may be used in combination with other aspects or combinations of aspects enumerated herein, at least one bistable valve maintains a state position by a beam, a magnet, or any drill geometry.

[0042] Other aspects of the present disclosure may be used in combination with other aspects or combinations of aspects recited herein.

[0043] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, at least one on-off valve maintains a state position according to the drill geometry.

[0044] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism controls media insertion into at least one biochip.

[0045] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism controls cell insertion into at least one biochip.

[0046] In another aspect of the disclosure, which may be used in combination with any other aspect or combination of aspects recited herein, at least one valve mechanism maintains cell culture by media insertion.

[0047] In another aspect of the disclosure, which may be used in combination with other aspects or combinations of aspects enumerated herein, a human-on-a-chip plate includes at least one 3D culture biochip, at least one microvalve, at least one microfluidic channel, and at least one inlet or outlet plate port. An outlet port from one chip may be connected to another chip by routing the valve. Multiple chips may be connected in parallel, series or combinations, and one or more chips in the human-on-a-chip plate may be bypassed. Each fluid connection may be independently changed at any time. At least one valve system may connect or separate different compartments of the organ-on-a-chip system. At least one inlet port and one outlet port access and block the microchannel. At least one valve system may extract, change the model flow map, and / or introduce or reduce at least one fluid chamber. The valve may be controlled to a bi-stable position by an actuator and a sensor and placed at a channel port of the chip. The valve may be placed right next to the biochip and controlled to a bi-stable position.

[0048] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the human-on-a-chip plate further comprises at least one organ-on-a-chip system including at least one ductal scaffold interconnecting with at least one surrounding compartment, the at least one organ-on-a-chip system including at least one ductal scaffold interconnecting with at least one surrounding compartment.

[0049] In another aspect of the present disclosure, which may be used in combination with other aspects or combinations of aspects enumerated herein, the plates may be quickly connected from the bottom or sides and fluids may be exchanged.

[0050] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the human-on-a-chip plate is replaceable, the ports on the human-on-a-chip align with the fittings on the biochip, and the valves are fixed or removable.

[0051] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, a human-on-a-chip plate includes integrated chips interconnected together.

[0052] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the valve is fixed or removable.

[0053] In another aspect of the present disclosure, which may be used in combination with any other aspect or combination of aspects enumerated herein, the human-on-a-chip allows access to the fluid from either the top or the side.

[0054] Another aspect of the present disclosure may be used in combination with other aspects or combinations of aspects enumerated herein, where a multiple logic microvalve system includes a plurality of microvalves, at least one fluid pathway including at least one channel, inlet and outlet ports, at least one bistable mechanism, at least one actuation mechanism, at least one structure surrounding other elements of the microvalve, where the fluid pathway has no dead volume, where the amount of actuation input is reduced by multiplexing between the multiple valves, and where the valves can control the fluid in the channel such that the flow is either hydrostatic or kinetic.

[0055] Another aspect of the present disclosure can be used in combination with any other aspect or combination of aspects enumerated herein, wherein at least one logic multiplexed microvalve is used to control fluid within a system.

[0056] Another aspect of the present disclosure may be used in combination with any other aspect or combination of aspects recited herein, where the valve may direct the sample to at least one extraction port.

[0057] Another aspect of the present disclosure can be used in combination with any other aspect or combination of aspects listed herein, where the bistable mechanism can be a bistable beam on a flexible tube valve.

[0058] Another aspect of the present disclosure may be used in combination with any other aspect or combination of aspects enumerated herein, where a fluid pathway may connect at least one biochip to multiple elements in a human-on-a-chip system.

[0059] Another aspect of the present disclosure may be used in combination with any other aspect or combination of aspects recited herein, where the bi-stable valve has two stable positions.

[0060] Another aspect of the present disclosure, which can be used in combination with any other aspect or combination of aspects enumerated herein, is where at least one channel includes a pneumatically actuated balloon.

[0061] Another aspect of the present disclosure can be used in combination with other aspects or combinations of aspects enumerated herein, where the system only requires power when switching between the two states, and once actuated, the bistable beam remains in place.

[0062] Another aspect of the present disclosure may be used in combination with any other aspect or combination of aspects listed herein, where the microvalve is a bi-stable magnetic drill piston valve.

[0063] Another aspect of the present disclosure may be used in combination with any other aspect or combination of aspects listed herein, where the bi-stable mechanism may include a magnetic channel.

[0064] Another aspect of the present disclosure may be used in combination with any other aspect or combination of aspects listed herein, where the bistable beam of the microvalve may be actuated either solenoidally or pneumatically.

[0065] In summary, the Human Operating System is a self-contained system that can grow and interconnect multiple cell types and run experiments automatically within the system. The purpose of this system is to be used for tissue modeling and drug testing to better predict the effects of diseases, biological parameters and drugs before testing in humans. It allows standardizing cell culture conditions to ensure an unprecedented level of physiological conditions. It also allows reliable running of real-time endpoint experiments within the system. The currently disclosed system allows an all-in-one approach that ensures standardization, reproducibility, precision, speed and cost reduction. [Brief description of the drawings]

[0066] Several exemplary apparatus embodiments of the present disclosure, and exemplary procedures for making and using one or more exemplary embodiments, are described in detail herein, by way of example, with reference to the accompanying drawings (which are not necessarily drawn to scale with respect to the internal or external structures illustrated), in which like reference characters indicate like elements throughout the several views.

[0067] [Figure 1] FIG. 1 shows a schematic diagram of two tubes (labeled I and IV) surrounded by a stroma (III) and contained within a housing (II), according to one example of the present disclosure. [Figure 2A] FIG. 2A shows a schematic diagram of one embodiment of a two vessel one stromal biochip according to one example of the present disclosure. [Figure 2B] FIG. 2B shows a schematic top view of one embodiment of a two vessel one stromal biochip according to one example of the present disclosure. [Figure 2C] FIG. 2C shows a schematic diagram of a two tube one stromal chip fabrication and assembly process according to one example of the present disclosure. [Figure 3A]FIG. 3A shows a schematic diagram of another embodiment of a 2-tube 1-stromal chip with three parallel channels according to one example of the present disclosure. [Figure 3B] FIG. 3B shows an exploded view of a 2-tube 1-stromal chip with three parallel channels according to one example of the present disclosure. [Figure 3C] FIG. 3C shows a schematic diagram of the fabrication and assembly process of one embodiment of a two-tube, one-stromal chip with three parallel channels according to one example of the present disclosure. [Figure 4A] FIG. 4A shows a schematic diagram of another embodiment of a 2-tube-1-stroma chip with three parallel channels with a stromal supply channel according to one example of the present disclosure. [Figure 4B] FIG. 4B shows detailed schematic top and bottom views of a 2-tube-1-stroma chip with three parallel channels with a stromal supply channel according to one example of the present disclosure. [Figure 4C] FIG. 4C shows an exploded view of one embodiment of a 2 tube 1 stromal tip with three parallel channels with a stromal supply channel according to one example of the present disclosure. [Figure 4D] FIG. 4D shows a schematic diagram of the fabrication and assembly process of a 2-tube-1-stroma chip with three parallel channels with a stromal supply channel according to one example of the present disclosure. [Figure 5A] FIG. 5A is a schematic diagram of a two-tube, one-stromal tip with different interstitial inlets according to one example of the present disclosure. [Figure 5B] FIG. 5B is a schematic top view of another embodiment of a two-tube, one-stromal tip having different interstitial inlets, according to one example of the present disclosure. [Figure 5C] FIG. 5C is an exploded view of a two tube one interstitial tip with different interstitial inlets according to one example of the present disclosure. [Figure 5D] FIG. 5D is a schematic diagram of a manufacturing and assembly process for a two-tube, one-stromal chip with different interstitial inlets according to an example of the present disclosure. [Figure 6A] FIG. 6A is a schematic diagram of a two tube one stroma chip having multiple stroma per tube according to one example of the present disclosure. [Figure 6B]FIG. 6B shows an exploded view of a 2 tube 1 stroma chip having multiple stroma per tube according to one example of the present disclosure. [Figure 6C] FIG. 6C shows a schematic diagram of the manufacturing and assembly process of a two tube one stroma chip with multiple stroma per tube according to one example of the present disclosure. [Figure 7A] FIG. 7A shows a schematic diagram of one embodiment of a human-on-biochip plate according to one example of the present disclosure. [Figure 7B] FIG. 7B shows a schematic diagram of an individual biochip element of a human-on-a-chip plate according to one example of the present disclosure. [Figure 7C] FIG. 7C illustrates an exploded view of a human-on-a-chip plate according to an example of the present disclosure. [Figure 8A] FIG. 8A shows a schematic diagram of one embodiment of a human-on-a-chip in which the individual chip entities are integrated as one piece within a housing and the valve mechanism connecting the individual chip elements is a cylindrical rotary mechanism, according to one example of the present disclosure. [Figure 8B] FIG. 8B shows a schematic diagram of an embodiment of a human-on-a-chip entity in which the individual chip entities are integrated as one part within a housing and the valve mechanism connecting the individual chip elements is a cylindrical rotary mechanism, according to an example of the present disclosure. [Figure 8C] FIG. 8C shows an exploded view of an embodiment of a human-on-a-chip in which the individual chip entities are assembled as one piece within the housing and the valve mechanism connecting the individual chip elements is a cylindrical rotary mechanism, according to an example of the present disclosure. [Figure 9A] FIG. 9A shows a schematic (isometric view) of one embodiment of a human-on-a-chip in which the individual chip entities are integrated as one piece within a housing and the valve mechanism connecting the individual chip elements is a pneumatically actuated flexible membrane mechanism, according to one example of the present disclosure. [Figure 9B]FIG. 9B shows a schematic diagram (top view) of another embodiment of a human-on-a-chip in which the individual chip entities are integrated as one piece within a housing and the valve mechanism connecting the individual chip elements is a pneumatically actuated flexible membrane mechanism according to one example of the present disclosure. [Figure 9C] FIG. 9C shows a schematic diagram (top view) of another embodiment of a human-on-a-chip in which the individual chip entities are integrated as one piece within a housing and the valve mechanism connecting the individual chip elements is a pneumatically actuated flexible membrane mechanism according to one example of the present disclosure. [Figure 9D] FIG. 9D shows a schematic diagram (top view) of one embodiment of the lower housing features of another embodiment of a human-on-a-chip in which the individual chip entities are integrated as one piece within the housing and the valve mechanism connecting the individual chip elements is a pneumatically actuated flexible membrane mechanism, according to one example of the present disclosure. [Figure 10A] FIG. 10A shows a schematic diagram of an embodiment of a top view of an embodiment of an HOC plate where the OOC is removable from the HOC plate and fluid is accessed from the side, according to an example of the present disclosure. [Figure 10B] 10B shows a schematic diagram of an embodiment of an exploded view of an embodiment of an HOC plate where the OOC is removable from the HOC plate and fluid is accessed from the side, according to an example of the present disclosure, and FIG 10C shows a schematic diagram of an embodiment of in-plate channeling of an embodiment of an HOC plate where the OOC is removable from the HOC plate and fluid is accessed from the side, according to an example of the present disclosure. [Figure 11] FIG. 11 shows a schematic diagram of one embodiment of a top view of one embodiment of an HOC plate where the OOC can be removed from the HOC plate and fluids and cells are extracted in situ according to one example of the present disclosure. [Figure 12] FIG. 12 shows a schematic diagram of one embodiment of a top view of one embodiment of an HOC plate where the OOC is removable from the HOC plate and fluids and cells are extracted through channels, chambers and valves, according to one example of the present disclosure. [Figure 13]FIG. 13 shows a schematic diagram of one embodiment of intra-plate channeling of one embodiment of a HOC plate where two tube chips are connected to one tube biochip according to one example of the present disclosure. [Figure 14] 14 shows a schematic diagram including pressure sensors at the inlet and outlet fluid ports of each chip to manipulate pressure within the channels of the chip, thereby controllably perfusing fluid from the tubular channels to the interstitial channels and vice versa, according to one example of the present disclosure. [Figure 15A] FIG. 15A shows a perspective view of a hydraulic valve mechanism connecting individual biochips according to one example of the present disclosure. [Figure 15B] FIG. 15B shows a perspective view of a cylindrical roll valve mechanism connecting individual biochips according to one example of the present disclosure. [Figure 15C] FIG. 15C shows a perspective view of a flexible membrane valve mechanism connecting individual biochips according to one example of the present disclosure. [Figure 15D] FIG. 15D shows a perspective view of a chip-connected microvalve assembly process according to one example of the present disclosure. [Figure 16A] FIG. 16A shows a perspective view of a direction-changing flow microvalve plate actuator according to one example of the present disclosure. [Figure 16B] FIG. 16B shows a detailed view of a redirection flow microvalve plate actuator according to one example of the present disclosure. [Figure 16C] FIG. 16C illustrates a perspective view of a solenoid magnet-gated microvalve according to one example of the present disclosure. [Figure 16D] FIG. 16D shows a detailed view of a solenoid magnet-gated microvalve according to one example of the present disclosure. [Figure 16E] FIG. 16E illustrates a perspective view of a solenoid magnet-gated microvalve plate according to one example of the present disclosure. [Figure 16F] FIG. 16F illustrates a detailed view of a solenoid magnet inlet outlet microvalve actuator plate according to an example of the present disclosure. [Figure 17A] FIG. 17A shows a perspective view of a normally-closed type pneumatically actuated on-off membrane microvalve according to one example of the present disclosure. [Figure 17B] FIG. 17B illustrates an exploded view of a normally-closed, pneumatically actuated on-off membrane microvalve according to one example of the present disclosure. [Figure 17C] FIG. 17C shows a perspective view of an additional embodiment of a normally open pneumatically actuated on-off membrane microvalve according to one example of the present disclosure. [Figure 17D] FIG. 17D illustrates an exploded view of an additional embodiment of a normally open pneumatically actuated on-off membrane microvalve according to one example of the present disclosure. [Figure 18A] FIG. 18A shows a perspective view of a pneumatically actuated bistable beam on a flexible tube microvalve according to an example of the present disclosure. [Figure 18B] FIG. 18B shows an exploded view of a pneumatically actuated bistable beam on a flexible tube microvalve according to one example of the present disclosure. [Figure 19] 19A and 19B show a perspective view and an exploded view, respectively, of a bi-stable magnetic drill piston microvalve according to one example of the present disclosure. [Figure 20A] FIG. 20A shows a perspective view of a bistable mechanically actuated flexible tube microvalve according to one example of the present disclosure. [Figure 20B] FIG. 20B shows a top view of one embodiment of a bistable mechanically actuated flexible tube microvalve with one bistable beam in a closed position according to one example of the present disclosure. [Figure 20C] FIG. 20C shows a cross-sectional view of a bistable mechanically actuated flexible tube microvalve in an open position according to one example of the present disclosure. [Figure 20D] FIG. 20D shows a cross-sectional view of a bistable mechanically actuated flexible tube microvalve in a closed position according to one example of the present disclosure. [Figure 21A] FIG. 21A shows a perspective view of a mechanically actuated ball-open microvalve according to one example of the present disclosure. [Figure 21B] FIG. 21B shows a close-up perspective view of one embodiment of a mechanically actuated ball-open microvalve (tip moving, screwdriver moving) according to one example of the present disclosure. [Figure 21C] FIG. 21C shows a detailed view of a socket that engages with a microvalve to switch when in an open or closed position, according to one example of the present disclosure. [Figure 21D] FIG. 21D shows a cross-section of a detailed view of a mechanically actuated ball-open microvalve according to one example of the present disclosure. [Figure 22A] FIG. 22A shows a cross-sectional view of a permanently sealed open / close microvalve in a closed position according to one example of the present disclosure. [Figure 22B] FIG. 22B shows a side view of a permanently sealed open / close microvalve in a preset closed position, with the pin firmly inserted into the channel to block the valve, according to one example of the present disclosure. [Figure 22C] FIG. 22C illustrates a cross-sectional view of a permanently sealed open / close microvalve in a preset open position according to one example of the present disclosure. [Figure 22D] FIG. 22D illustrates a side view of a permanently sealed open / close microvalve in an open position, with the pin not fully inserted into the channel, according to one example of the present disclosure. [Figure 23A] FIG. 23A illustrates a perspective view of one embodiment of an assembled operating system according to an example of the present disclosure. [Figure 23B] FIG. 23B illustrates a perspective view of one embodiment of the major components of an assembled operating system according to one example of the present disclosure. [Figure 23C] FIG. 23C illustrates a perspective view of a top view of one embodiment of a fluidic component of an assembled operating system, according to one example of the present disclosure. [Figure 23D] FIG. 23D illustrates a perspective view of one embodiment of an assembled operating system with the mixing chamber drawer open according to one example of the present disclosure. [Figure 23E] FIG. 23E illustrates an exploded view of one embodiment of an operating system according to an example of the present disclosure. [Figure 23F] FIG. 23F illustrates a side view of one embodiment of a pressure-driven pump for a microchamber of an assembled operating system according to one example of the present disclosure. [Figure 23G] FIG. 23G shows a bottom perspective view of one embodiment of a pressure-driven pump for a microchamber of an assembled operating system according to one example of the present disclosure. [Figure 23H] FIG. 23H shows a bottom perspective view of one embodiment of a pressure-driven pump for a microchamber of an assembled operating system according to one example of the present disclosure. [Figure 23I] FIG. 23I shows a bottom perspective view of one embodiment of a pressure-driven pump for a microchamber of an assembled operating system according to one example of the present disclosure. [Figure 23J] FIG. 23J illustrates a bottom perspective view of one embodiment of a pressure-driven pump for a microchamber of an assembled operating system according to one example of the present disclosure. [Figure 23K] FIG. 23K illustrates a perspective view of one embodiment of a flow diagram for an assembled operating system according to an example of the present disclosure. [Figure 23L] FIG. 23L shows a flow diagram of a 1 to 72 distributor connecting a mixing chamber to a media chamber connected to a human-on-a-chip plate according to one example of the present disclosure. [Figure 24] FIG. 24 illustrates a flow diagram of another embodiment of an assembled operating system according to an example of the present disclosure. [Figure 25A] FIG. 25A illustrates a perspective view of another embodiment of an operating system cover according to an example of the present disclosure. [Figure 25B] FIG. 25B illustrates an exploded view of an operating system cover according to an example of the present disclosure. [Figure 25C] FIG. 25C illustrates a perspective view of a human chip insertion process within an operating system according to one example of the present disclosure. [Figure 25D] FIG. 25D illustrates a perspective view of the insertion of a spectroscopic probe and sensing plate into an operating system according to one example of the present disclosure. [Figure 25E] FIG. 25E illustrates an exploded view of all the elements of an operating system according to one example of the present disclosure. [Figure 25F] FIG. 25F illustrates a perspective view of a fluid control of an operating system according to an example of the present disclosure. [Figure 26A] FIG. 26A illustrates a perspective view of one embodiment of a pneumatic valve actuated operating system according to one example of the present disclosure. [Figure 26B] FIG. 26B illustrates an exploded view of a pneumatic valve actuated operating system according to an example of the present disclosure. [Figure 26C] FIG. 26C illustrates a top view of a pneumatic valve actuated operating system according to an example of the present disclosure. [Figure 27A] FIG. 27A shows an embodiment of the assembly of the media chamber of another embodiment of a human-on-a-chip in which the individual chip entities are integrated as one piece within the housing and the valve mechanism connecting the individual chip elements is a pneumatically actuated flexible membrane mechanism (isometric view). [Figure 27B] Figure 27B shows an exploded assembly of the media chamber of another embodiment of a human-on-a-chip in which the individual chip entities are assembled as one piece within the housing and the valve mechanism connecting the individual chip elements is a pneumatically actuated flexible membrane mechanism (isometric view). [Figure 27C] FIG. 27C shows a perspective view of a media chamber plate according to one example of the present disclosure. [Figure 27D] FIG. 27D shows an exploded view of a media chamber plate according to one example of the present disclosure. [Figure 27E] FIG. 27E illustrates a perspective view of a piston plate according to an example of the present disclosure. [Figure 27F] FIG. 27F illustrates a perspective view of an additional embodiment of a piston plate according to an example of the present disclosure. [Figure 27G] FIG. 27G illustrates a side view of one embodiment of a piston plate according to an example of the present disclosure. [Fig. 27H] FIG. 27H shows a detailed view of a single piston of FIGS. 27E-27G, according to an example of the present disclosure. [Figure 27I]FIG. 27I illustrates a top perspective view of a piston actuator plate according to an example of the present disclosure. [Figure 27J] FIG. 27J illustrates a bottom perspective view of a piston actuator plate according to an example of the present disclosure. [Figure 27K] FIG. 27K shows a perspective view of a fluid source media plate according to one example of the present disclosure. [Figure 28A] FIG. 28A illustrates a flow diagram of a human-on-chip operating system process according to one example of the present disclosure. [Figure 28B] FIG. 28B illustrates a flow diagram of a media chamber filling process of a human-on-chip operating system according to an example of the present disclosure. [Figure 28C] FIG. 28C illustrates a flow diagram of a media chamber ejection process of a human-on-chip operating system according to one example of the present disclosure. [Figure 28D] FIG. 28D illustrates a flow diagram of a media chamber seeding process of a human-on-chip operating system according to one example of the present disclosure. [Figure 28E] FIG. 28E illustrates a flow diagram of a media sample collection process of a human-on-a-chip operating system according to one example of the present disclosure. [Figure 28F] FIG. 28F illustrates a flow diagram of a cell sample collection process of a human-on-a-chip operating system according to an example of the present disclosure. [Figure 29A] FIG. 29A shows a flow diagram of fluid control in a triple measurement mechanism according to one example of the present disclosure. [Figure 29B] FIG. 29B shows a logic control diagram of fluid control for FTIR extraction according to one example of the present disclosure. [Figure 30A] FIG. 30A shows a flow diagram for delivering a sample from a media chamber to a biochip according to one example of the present disclosure. [Figure 30B] FIG. 30B shows a flow diagram of delivering a sample from a biochip to a measurement device according to one example of the present disclosure. [Figure 31A]FIG. 31A shows a flow diagram of gel delivery to an interstitial channel according to one example of the present disclosure. [Figure 31B] FIG. 31B shows a flow diagram of pumping gel out of an interstitial channel according to one example of the present disclosure. [Figure 32A] FIG. 32A shows a schematic diagram of a fully assembled embodiment of a mixing chamber according to one example of the present disclosure. [Figure 32B] FIG. 32B shows a schematic diagram of an embodiment of a cell source mixing chamber according to one example of the present disclosure. [Figure 32C] FIG. 32C shows a schematic diagram of an embodiment of a media source mixing chamber according to one example of the present disclosure. [Fig. 32D] FIG. 32D shows a schematic diagram of a cell chamber and a media chamber side-by-side according to one example of the present disclosure. [Figure 32E] FIG. 32E illustrates a flow diagram of one embodiment of a bubble filter and pressure sensor in a mixing chamber according to one example of the present disclosure. [Figure 33A] FIG. 33A shows a flow diagram of one embodiment of a mixing chamber with all 36 tubes exiting the mixing system according to one example of the present disclosure. [Figure 33B] FIG. 33B shows a perspective view of an embodiment of a mixing chamber where all 36 tubes enter into only 3 tubes exiting the mixing system according to one example of the present disclosure. [Figure 33C] FIG. 33C shows a flow diagram of one embodiment of a mixing chamber where all 36 gas tubes are connected to the cell chamber according to one example of the present disclosure. [Figure 33D] FIG. 33D shows a flow diagram of one embodiment of a mixing chamber where all three pressure valves are connected to a cell chamber according to one example of the present disclosure. [Figure 33E]FIG. 33E shows a flow diagram of one embodiment of a mixing chamber according to one example of the present disclosure, where each row of cell chambers is connected to a 1 / 12-way valve that can switch between each row of cell chambers to route the sample through a flow sensor, air filter, and pressure gauge connected to a quick connect that connects the mixing chamber to the human-on-chip plate. [Figure 33F] FIG. 33F shows a flow diagram of a single cell chamber entity according to one example of the present disclosure, where the cell chamber is connected to pressure and gas from one side and to a 1 / 12 way valve from the other side that is temporarily connected to a sensor, air filter, and a pressure gauge that is connected to a quick connect connecting the mixing chamber and the human-on-chip plate. [Figure 33G] FIG. 33G shows a flow diagram of one embodiment of a mixing chamber with all 12 tubes connected to a media chamber according to one example of the present disclosure. [Fig. 33H] FIG. 33H shows a flow diagram of one embodiment of a mixing chamber where all three pressure valves are connected to a media chamber according to one example of the present disclosure. [Figure 33I] FIG. 33I shows a flow diagram of one embodiment of a mixing chamber according to one example of the present disclosure, where each row of media chambers is connected to a 1 / 4-way valve that can switch between the rows of media chambers to route the sample through a flow sensor, air filter, and pressure gauge connected to a quick connect that connects the mixing chamber to the human-on-a-chip plate. [Figure 33J] FIG. 33J shows a flow diagram of a single media chamber entity according to one example of the present disclosure, where the media chamber is connected to pressure and gas from one side and to a 1 / 4-way valve from the other side that is temporarily connected to a sensor, air filter, and a pressure gauge that is connected to a quick connect connecting the mixing chamber and the human-on-chip plate. [Figure 34A]FIG. 34A shows a flow diagram of one embodiment of a mixing chamber with access to O2, CO2 and N2 bottles, as well as O2 and CO2 sensors, all located below the cell chamber, according to one example of the present disclosure. [Figure 34B] FIG. 34B shows a diagram of one embodiment of an O2 and CO2 sensor at the tissue level grown within a chip, according to one example of the present disclosure. [Figure 35A] FIG. 35A shows an exploded view of a heat exchange plate passing temperature-controlled fluid in contact with a human-on-chip according to one example of the present disclosure. [Figure 35B] FIG. 35B shows a schematic diagram of a heat exchange generator chamber that heats and cools a fluid coming to the heat exchange plate according to one example of the present disclosure. [Figure 36A] FIG. 36A shows a flow diagram of one embodiment of a mechanical actuation mechanism that directs fluid through the system while avoiding clogging of channels and damage to valves, actuators, or sensors, according to one example of the present disclosure. [Figure 36B] FIG. 36B shows a flow diagram of an embodiment of a mechanical triggering mechanism to target valves for temperature, O2 and CO2 levels while cleaning damaged pre-cell seeding systems, actuators or sensors, according to one example of the present disclosure. [Figure 36C] FIG. 36C shows a flow diagram of one embodiment of a mechanical actuation mechanism that uses pressure sensors and is assisted by microscopic computer vision to ensure there are no air bubbles in the system (all liquid pressurization, imaging, optical contrast, fluorescence) prior to cell seeding, according to one example of the present disclosure. [Figure 36D] FIG. 36D shows a flow diagram of one embodiment of machine-initiated detection and display of fluid flow, non-wetted surfaces, and air bubbles using thermal imaging (flash thermography), according to one example of the present disclosure. [Figure 36E] FIG. 36E shows a flow diagram of one embodiment of the machine start and run mechanism showing temperature, O2, CO2, pressure sensors, as well as fluid channeling, CO2 and O2 mixing, bubble traps, and a heating plate that controls all parameters of the system according to one example of the present disclosure. [Figure 37A] FIG. 37A shows a perspective view of an embodiment of a multi-lens microscope and spectroscopy setup according to one example of the present disclosure. [Figure 37B] FIG. 37B shows a perspective view of an embodiment of a microscope and spectroscopy setup with one lens moving in the xy plane according to one example of the present disclosure. [Figure 37C] FIG. 37C shows a perspective view of an embodiment of a multiple sensor FTIR readout setup according to one example of the present disclosure. [Figure 37D] FIG. 37D shows a perspective view of an embodiment of an FTIR readout setup with only one sensor where fluid is extracted and measured while a time interval is calculated according to one example of the present disclosure. [Figure 37E] FIG. 37E shows a perspective view of a detailed extraction mechanism of one embodiment of an FTIR reading mechanism with only one sensor where fluid is extracted and measured while a time interval is calculated according to one example of the present disclosure. [Figure 37F] FIG. 37F shows a perspective view of an embodiment of an FTIR readout setup with a flow cell integrated into the chip according to one example of the present disclosure. [Figure 38A] FIG. 38A shows a top view of a possible biochip layout for human-on-chip modeling, according to one example of the present disclosure. [Figure 38B] FIG. 38B shows a projection of a possible biochip layout for human-on-a-chip modeling, according to one example of the present disclosure. [Figure 38C] FIG. 38C shows a top view of an additional embodiment of a possible biochip configuration for human-on-a-chip modeling, according to one example of the present disclosure. [Figure 38D] 38D shows a perspective view of an additional embodiment of a possible biochip arrangement for cell tissue modeling according to one example of the present disclosure. Briefly, a tubular channel (II) containing endothelial or epithelial cells connects one biochip (I) to at least another biochip (III). [Figure 39A]39A shows a cross-sectional view of an embodiment of a two-tube chip tissue tubular structure according to one example of the present disclosure, where I. is the interstitial channel of the biochip, II. is the extracellular matrix, III. is endothelial and / or epithelial cells, and may also contain cancer cells, IV. is the tubular channel containing media, and V. is the interstitial channel containing cells, which may include cancer cells. [Figure 39B] FIG. 39B is a perspective view of an embodiment of multiple cultured tissues in a chip connected to a common tube / vessel according to one example of the present disclosure. [Figure 39C] FIG. 39C is a schematic diagram of an embodiment of an organ-on-chip grown on a human-on-chip according to one example of the present disclosure. The cells can be primary cells, cell lines, IPSCs, patient-derived cells, or biopsies and can be used for precision medicine analysis or more general analysis. I. is the interstitial channel of the biochip. II. is the extracellular matrix. III. is tissue or tumor cells of interest. IV. is immune cells. V. is endothelial and / or epithelial cells. VI. is the tubular channel of the biochip. [Figure 39D] 39D shows a schematic diagram of an embodiment of a liquid-air interface lung-on-a-chip grown in a human-on-a-chip according to one example of the present disclosure, where I. is the interstitial channel of the biochip, II. is the extracellular matrix, III. represents endothelial cells, IV. is the tubular channel of the biochip containing air, V. represents lung tissue, VI. is the respiratory tract, and VI. are astrocytes. [Figure 39E] 39E shows a schematic diagram of one embodiment of a glioblastoma model and blood-brain barrier in a biochip according to one example of the present disclosure, where I. is the interstitial channel of the biochip, II. is the extracellular matrix, III. represents glioblastoma cells with immune cell infiltration, IV. is immune cells, V. is endothelial cells and pericytes, VI. is the tubular channel of the biochip, and VI. is astrocytes. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0068] The present disclosure provides five aspects that enable accurate, repeatable, semi-automated, precise and predictive experimental systems. The provided experiments allow for precise creation and maintenance of tissue models, drug testing, real-time extraction and measurement, and software-assisted control and data analysis. The provided systems allow for more accurate prediction of drug toxicity, efficacy and dosing, making them a powerful tool for precise personalized drug prognosis.

[0069] A first aspect of the present disclosure provides a biomimetic tissue culture platform that enables fundamental aspects of a biomimetic model. The biochip provided embeds multiple cylindrical porous tubular scaffolds that form different compartments within the biochip.

[0070] A second aspect of the present disclosure provides a hybrid human-on-chip tissue culture platform for human models, which can be used alone or with the aid of an operating system.

[0071] A third aspect of the present disclosure provides a semi-automated fluid control system of valves and pumps that can manipulate and control very small volumes to move samples throughout all areas of the system and to move samples in and out of the system.

[0072] A fourth aspect of the present disclosure provides a semi-automated sample collection and real-time measurement add-on, where the configuration of the system allows interfaces, microscopes, spectroscopy, Fourier transform infrared (FTIR), and other measurement devices to measure all samples while the experiment is being performed and pumped in and out of the system.

[0073] A fifth aspect of the present disclosure provides software for automating and controlling the operating system, the software including different commands for different experimental setups, and further, the software is connected to a measurement device and allows for performing data acquisition that acts on the measurement device and the fluid control mechanism of the system.

[0074] Multi-duct biochip

[0075] In one embodiment, the biochip includes at least two ducts and one stroma, where the ducts are formed from a cylindrical ultra-thin porous membrane that does not need to be curved. In additional embodiments, the ducts are made from Gore flexible tubing or made from any material, and the holes are formed by laser gunning or extrusion. In one embodiment, the biochip may include multiple ducts and multiple stroma. Additionally, a different solution may be inserted into the stroma than the solution passing through the multiple ducts. In additional embodiments, known materials such as stents may be utilized in the multiple biochips.

[0076] In one embodiment, the tubes can be fabricated utilizing separate hydrophobic and hydrophilic membranes. In additional embodiments, one of the tubes can be formed from a single membrane with different regions that are treated to be hydrophobic or hydrophilic by plasma or UV treatment or coating. Depending on the particular requirements of the system user, the hydrophobic and hydrophilic properties of the tubes can be tailored according to known treatment techniques.

[0077] In one embodiment, the biochip may contain multiple cavities, allowing for the cultivation of multiple organs on a single multi-channel biochip.

[0078] In one embodiment, a biochip with a single tube may also be utilized with the presently disclosed system. For example, in one embodiment, the biochip utilized is the biochip described in PCT Application No. PCT / QA2021 / 050016 entitled "DUCT ORGANOID-ON-CHIP," filed June 25, 2021, which is assigned to the assignee of the present invention and is incorporated herein by reference in its entirety, and which describes a biomimetic tissue culture platform or biochip that enables fundamental aspects of a biomimetic model.

[0079] FIG. 1 shows a schematic diagram of two vessels surrounded by a stroma according to one example of the present disclosure.

[0080] 2A shows a perspective view of a two-tube, one-stromal biochip according to one example of the present disclosure. In one embodiment, the housing of the biochip is constructed of a material that does not impede optical or spectroscopic access to the multiple tubes.

[0081] FIG. 2B shows a top view of a further embodiment of a two-tube, one-stromal biochip according to one example of the present disclosure. In one embodiment, the biochip includes two porous cylindrical tubular channels surrounded by the same interstitial chamber, and an air filtration region for each tube to release air bubbles from the biochip. In some examples, the air filtration region also allows air to enter the biochip, thereby making air accessible to cells in culture. In some examples, the biochip includes an inlet and outlet for each tube, and further includes an inlet and outlet for the interstitial channel. The biochip may also include multiple segments of tubing surrounded by plastic. In one example, the biochip includes a cover glass on the top of the biochip to cover at least a portion of the top surface of the top housing of the biochip. The top cover glass may be a thin cover glass made of a brittle, transparent, low autofluorescence material such as glass or a polymer. The top and bottom housings include features such that when placed in direct contact, an interior space is formed to accommodate a porous tube scaffold made of a membrane. These features may be the result of engraving into the inner layer of the housing. In another example, the upper and lower housings contain features that form microfluidic channels that connect to the external compartments of the tube scaffold, and these features are engraved into the exterior surface of the housing and covered with another housing part or a thin cover glass to create a complete channel in the lower housing that can accommodate multiple porous membrane structures.

[0082] In one embodiment, the upper and lower housings include features that hold the cylindrical tube scaffold in a position that allows access to the respective internal and external compartments of the tube scaffold. The housings that form the microfluidic channels that connect to the internal compartments of the tube scaffold can extend beyond the inlet and outlet holes of the channels, which can be later blocked using plugs after assembly of the biochip. A bottom cover glass similar to the top cover glass can be provided to cover the bottom of the bottom housing. In one example, the cover slip glasses that form the top and bottom layers of the chip are bonded to the housings using a glass polymer adhesive. In one example, the porous membrane structure is formed by bending the porous membrane by rotating it 180 degrees to form a cylindrical structure and bonding the access material of the unbent membrane to the cylindrical structure. For example, two pre-bonded cylindrical hydrophobic ultra-thin porous membranes on each tube can be used for filtering. These elements can be combined to form multiple porous membrane tube structures surrounded by the same stroma.

[0083] FIG. 2C shows a flow diagram of the fabrication and assembly process of a two tube one interstitial biochip according to one example of the present disclosure. When bending and tensioning of the membrane are performed simultaneously during bonding, the porous membrane is bent into a cylindrical shape and in a direction of about 180°. In one example, a biochip constructed according to this method can include an air filtration region, a tube surrounded by stroma, a tube inlet, a tube outlet, an interstitial channel inlet, and an interstitial channel outlet. FIG. 2C shows steps and processes for performing a bonding process of biochip elements in which the tube scaffold is bent, tensioned, and bonded to the housing at its ends according to this method. For example, this method includes placing a hydrophobic membrane and a hydrophilic membrane on the housing, bending it over a rod, and bonding the ends of the membrane to the housing using adhesive, heat, or chemicals. After placing the membrane on the housing, bending the membrane over two rods that form each of the two tubes, tensioning the membrane, and bonding the ends of the membrane to the housing, a pin is assembled into the outlet interstitial hole of the lower housing. Then, everything is sandwiched between two housings and bonded using chemically assisted thermocompression. Finally, the method includes a rod and pin removal process, plugging the ends of both tubular channels beyond the inlet and outlet holes, and assembling cover slip glass on both sides of the biochip.

[0084] 3A shows a perspective view of an embodiment of a two-tube, one-stromal biochip according to one example of the present disclosure. In one embodiment, the biochip includes two porous cylindrical tubular channels surrounded by the same interstitial chamber with a stromal supply channel, and the upper tube can be used for insertion of gel into the interstitial chamber.

[0085] 3B shows a perspective view of a further embodiment of a two tube one stroma biochip with a stroma supply channel according to one example of the present disclosure. In one embodiment, the biochip may include two porous cylindrical tubular channels surrounded by the same stroma chamber with a stroma supply channel, where the stroma supply channel is a perforated tube allowing direct supply to the stroma, and the wide pores in the tube increase the diffusion rate between the tube and the stroma.

[0086] FIG. 3C shows a flow diagram of the fabrication and assembly process of a two tube one interstitial biochip with three parallel channels according to one example of the present disclosure. The upper and lower housings contain three parallel channels holding two cylindrical tube scaffolds and one perforated tube, and one interstitial inlet located to provide access to the interior and exterior compartments of each tube scaffold. In one embodiment, the perforated tube is glued at the end of the middle channel, the tube is formed from an ultra-thin porous membrane curved into a cylindrical shape around a rod and glued to the housing at the end using adhesive, heat, or chemical means, and a pin is assembled into the outlet interstitial hole of the lower housing. Then, sandwich everything between the two housings and bond them together using chemically assisted thermocompression. Finally, the method may include a removal process of the rod and pin, plugging the ends of both tubular channels beyond the inlet and outlet holes, and assembling a cover slip glass on both sides of the biochip.

[0087] Figure 4A shows another perspective view of a two-tube one-stroma biochip with three parallel channels according to an example of the present invention. In one embodiment, two tubes with porous membranes are surrounded by one stroma, the upper central tube is connected to the stroma chamber, and these two tubes can be used for feeding.

[0088] 4B shows a perspective view of a two tube one stroma biochip with three parallel channels according to one example of the present disclosure. In one embodiment, two tubes with porous membranes are surrounded by one stroma, allowing two types of endothelial cells to be cultured in the tubes and one type of epithelial cells to be cultured in the stroma.

[0089] 4C shows a perspective view of a further embodiment of a two tube one interstitial biochip with three parallel channels with an interstitial supply channel according to one example of the present disclosure. In one embodiment, the biochip may include two porous cylindrical tubular channels surrounded by the same interstitial chamber with an interstitial supply channel, the interstitial supply channel being a perforated tube allowing direct supply to the interstitium, and wide pores in the tube increasing the diffusion rate between the tube and the interstitium.

[0090] FIG. 4D shows a flow diagram of the fabrication and assembly process of a two tube one interstitial biochip with three parallel channels according to one example of the present disclosure. The upper and lower housings contain three parallel channels that hold two cylindrical tube scaffolds and one interstitial inlet in a position that allows access to the interior and exterior compartments of each tube scaffold. In one embodiment, the tubes are formed from an ultra-thin porous membrane curved into a cylindrical shape around a rod and attached to the housing at its ends using adhesive, heat, or chemical means, and a pin is assembled into the outlet interstitial hole of the lower housing. Then, everything is sandwiched between the two housings and they are bonded using chemically assisted thermocompression. Finally, the method may include a removal process of the rod and pin, plugging the ends of both tubular channels beyond the inlet and outlet holes, and assembling a cover slip glass on both sides of the biochip.

[0091] 5A shows a perspective view of a two-tube-one-stromal chip with different interstitial inlets according to one example of the present disclosure. In one embodiment, the biochip includes two porous cylindrical tubular channels surrounded by the same interstitial chamber with different interstitial inlets, allowing different types of cell organs to be cultured within the same tube on the two-tube-one-stromal biochip.

[0092] 5B shows a top view of a two-tube-one-stroma chip with different stromal inlets according to one example of the present disclosure. In one embodiment, the biochip includes two porous cylindrical tubular channels surrounded by the same stromal chamber with different stromal inlets, each of which can be used to inject gel into the stroma connected to the inlet.

[0093] 5C shows a perspective view of a two tube one stromal chip with different stromal inlets according to one example of the present disclosure. In one embodiment, the biochip includes two porous cylindrical tubular channels surrounded by the same stromal chamber with different stromal inlets, the stromal chamber having a perforated barrier dividing it into two separate stroma.

[0094] FIG. 5D shows a flow diagram of the fabrication and assembly process of a 2 tube 1 stromal chip with different interstitial inlets according to one example of the present disclosure. The upper and lower housings contain two cylindrical tube scaffolds and three parallel channels that hold two different interstitial inlets for each interstitial chamber in a position that allows access to the interior and exterior compartments of each tube scaffold. In one embodiment, the interstitial chamber is divided into two separate stroma by a perforated barrier that is glued at the end of the central channel. The tubes are formed from an ultra-thin porous membrane curved cylindrically around a rod and glued to the housing at the end using adhesive, heat, or chemical means, and a pin is assembled into the outlet interstitial hole of the lower housing. Then, sandwich everything between the two housings and bond them together using chemically assisted thermocompression. Finally, the method may include a rod and pin removal process, plugging the ends of both tubular channels beyond the inlet and outlet holes, and assembling a cover slip glass on both sides of the biochip.

[0095] 6A shows a perspective view of a multi-stroma per tube biochip with three stroma and two tubes according to one example of the present disclosure. In one embodiment, the biochip can include multiple cavities to allow multiple types of cell organs to be cultured in the same tube on a single multi-stroma per tube biochip.

[0096] 6B shows a perspective view of a multi-stroma per tube biochip with three stroma and two tubes according to an example of the present disclosure. In one embodiment, the biochip can include multiple cavities to allow the same type of cell organelles to be cultured in the same tube on a single multi-stroma per tube biochip.

[0097] FIG. 6C shows a flow diagram of the manufacturing and assembly process of a biochip with three stroma and two tubes with multiple stroma per tube according to one example of the present disclosure. The upper and lower housings contain two cylindrical tube scaffolds and three parallel channels that hold one stroma inlet for each stroma chamber at a position that allows access to the interior and exterior compartments of each tube scaffold. In one embodiment, the tubes are formed from an ultra-thin porous membrane curved cylindrically around a rod and attached to the housing at its end using adhesive, heat, or chemical means, and a pin is assembled into the outlet stroma hole of the lower housing. Then, the whole is sandwiched between the two housings and they are bonded using chemically assisted thermocompression. Finally, the method may include a removal process of the rods and pins, plugging the ends of both tubular channels beyond the inlet and outlet holes, and assembling a cover slip glass on both sides of the biochip.

[0098] HUMAN-ON-CHIP PLATE

[0099] In one embodiment, the human-on-a-chip plate includes tubes of cylindrical cross-section surrounded by stroma such that blood vessel ducts can pass through the stroma of various tissues, including epithelial ducts that pass through individual stroma.

[0100] In additional embodiments, the biochip plate uses valves between individual biochips and elements, allowing the valves to be opened and closed manually or automatically via an operating system. In one embodiment, the present disclosure is a hybrid controlled organ-on-a-chip.

[0101] Human-on-a-chip plates can be made by piecing together different individual chips like a puzzle by interconnecting the different chips through their tubing to the inlet / outlet valves of the biochip and valve-holding plate. Human-on-a-chip plates can also incorporate different chips in the same housing, and instead of being separate elements with valves, they can be assembled within the biochip as other elements are being assembled. The valves connecting the different tissue culture channels of the biochip can be hydraulic piston mechanisms, valves, hydraulic rolling or pneumatically actuated membrane piston valves, or simple membrane deformation valves. Any type of valve can be incorporated into this system.

[0102] The human-on-a-chip plate may also include normally closed on-off valves and waste removal channels built into the biochip for use when assembled in the operating system. The cover glass covering the individual chip units may be made of one or more pieces of glass covering the surface channels of each entity, or alternatively only one piece of glass may be used to cover the entire surface of the biochip. The interconnection between the different biochip entities may be made through tube openings that run straight along their axis, or they may connect to the surrounding biochip through inlet / outlet holes. The interface of the biochip to the operating system may be through the same inlet / outlet holes used for manual use, or through another hole separated from the system by a normally closed valve actuated by the operating system.

[0103] In one embodiment, the presently disclosed system includes a means for rotating the human-on-a-chip plate about a vertical and / or horizontal axis to ensure uniform distribution of reagents within the solutions, and uniform distribution and attachment of cells after seeding into the channels.

[0104] In one embodiment, the human-on-a-chip plate allows individual biochips to be connected together to form a series of interconnected biochips. Additionally, in one embodiment, specific biochips can be selectively disconnected from the remaining interconnected biochips to create multiple biochip circuits. As described later in this disclosure, adjustment of the fluid control mechanism can open and close specific valves to facilitate multiple combinations of interconnected biochip circuits. In this disclosure, the modularity of the human-on-a-chip plate allows for dynamic delivery of solutions to specific biochips, thereby allowing multiple protocols to be run in parallel within a single human-on-a-chip plate. Dynamic delivery of solutions in the system of the present disclosure allows specific biochips to be isolated from the remaining biochips to run precision medicine protocols.

[0105] In one embodiment, the human-on-a-chip fluidly connects multiple chips to each other and to other elements of the operating system. These elements may be, but are not limited to, other chips, waste chambers, or extraction chambers. In one embodiment, the connections can be changed at any time.

[0106] 7A shows a perspective view of a human-on-a-chip plate and individual chips according to one example of the present disclosure. In one embodiment, the individually removable biochips are assembled together by interconnecting the different chips via their tubing to the inlet / outlet valves of the biochip and valve holding plate.

[0107] 7B shows a perspective view of an individual chip in a human-on-a-chip plate according to one embodiment of the present disclosure. In one embodiment, the individually removable biochips are assembled in a puzzle-like fashion by interconnecting different chips through their tubes to inlet / outlet valves on the biochip and valve-holding plate, and fluids are pumped from the top of the human-on-a-chip plate through the tubes and from one tube to another through a cylindrical rotating valve mechanism.

[0108] 7C shows an exploded view of a human-on-a-chip plate according to one embodiment of the present disclosure. In one embodiment, the individually removable biochips are assembled in a puzzle-like fashion by interconnecting different chips through their tubes to inlet / outlet valves on the biochip and valve-holding plate, and fluids are pumped from the top of the human-on-a-chip plate through the tubes and from one tube to another through a cylindrical rotating valve mechanism.

[0109] 8A and 9A show perspective views of a human-on-a-chip plate according to one embodiment of the present disclosure. In one embodiment, the individual biochip entities are integrated into a human-on-a-chip plate, with a cylindrical rotation mechanism connecting the housing and a valve mechanism connecting the individual chip elements.

[0110] 8B and 9B show detailed views of the valve mechanism connecting the individual biochip elements of the human-on-chip plate according to an example of the present disclosure. In one embodiment, the individual biochip entities are integrated as an integrated human-on-chip. Specifically, a cylindrical rotating mechanism connects the housing with the valve mechanism connecting the individual biochip elements.

[0111] 8C and 9C show exploded views of a human-on-a-chip plate according to one embodiment of the present disclosure. In one embodiment, the individual biochip entities are integrated into a housing and valve mechanism as one element and connected via a cylindrical rotation mechanism.

[0112] FIG. 10A shows a detailed view of a human-on-a-chip plate according to an example of the present disclosure. In one embodiment, the individually removable biochips are assembled in a puzzle-like fashion by interconnecting different chips through their tubes to the biochips and inlet / outlet valves on the valve-holding plate. The biochip is made up of several plate layers stacked on top of each other and is composed of a heating plate, a chip-holding plate, a chip-attaching plate, a tube 1 fluid channeling plate, an interstitial fluid channeling tube, a tube 2 fluid channeling plate, a row actuation surface, a column 1 actuation plate, a column 2 actuation plate, and two chip actuation plates.

[0113] The figure shows a schematic diagram of a human-on-a-chip plate according to one example of the present disclosure. In one embodiment, the heating plate includes fluid channels that run through the inlets and outlets on top of the chip and are fitted with an operating system that can pump any liquid at any temperature to this heating plate to control, increase, decrease, or maintain the temperature of the cell culture.

[0114] FIG. 10C shows an exploded view of a human-on-a-chip plate according to an example of the present disclosure. In one embodiment, the row actuation plate is a valve actuation channel plate connected to the first bistable beam of each valve in the same row (3 rows, 3 signals). The column 1 actuation plate is a valve actuation channel plate connected to the second bistable beam of each valve in the first two columns (2 columns, 2 signals). The column 2 actuation plate is a valve actuation channel plate connected to the second bistable beam of each valve in the second two columns (2 columns, 2 signals). The two chip actuation plates are valve actuation channel plates connected to the third bistable beam of each valve in each chip (2 tubes and 1 interstitium, 3 signals). In some cases, the bistable valves can open two bistable beams to each other. For example, to open the valve connecting tube 1 of chip 1 in row 1 column 1 to tube 1 of chip 2 in row 1 column 2, the signal would be (1,1,1), which would open the first pneumatic tubes in the first, second, and fourth layers.

[0115] 11A shows a perspective view of a human-on-a-chip plate according to one example of the present disclosure. In one embodiment, the individually removable biochips are combined by interconnecting the different chips via their tubing to the inlet / outlet valves of the biochip and valve-holding plate, and fluids are extracted from specific extraction locations within each biochip of the human-on-a-chip plate. A pipette-like mechanism can be used or implemented in the system to take the samples.

[0116] 12A shows a perspective view of a human-on-a-chip plate according to one embodiment of the present disclosure. In one embodiment, individually removable biochips are combined by interconnecting different chips through their tubing to the inlet / outlet valves of the biochip and valve holding plate, samples are collected in situ through channels, chambers, and valves, and cell samples can be collected in situ using a pipette-like mechanism or pumped through a valve mechanism to an extraction chamber or media chamber.

[0117] In one embodiment, the individual biochip entities are incorporated as a human-on-chip plate with a cylindrical rotating mechanism connecting the housing and a valve mechanism connecting the individual chip elements, and fluids are pumped from the top of the human-on-chip plate through tubes and from one tube to another through a valve system.

[0118] 13A shows a perspective view of a human-on-a-chip plate according to one embodiment of the present disclosure. In one embodiment, the individual biochip entities are assembled into a single human-on-a-chip plate, with a cylindrical rotating mechanism connecting the housing and a valve mechanism connecting the individual chip elements, and fluids are pumped through tubes on the side of the human-on-a-chip plate and from tube to tube via a valve system.

[0119] In one embodiment, the individual biochip entities are integrated into an integrated human-on-chip: a cylindrical rotating mechanism connects the housing with a valve mechanism that connects the individual biochip elements, and fluids are pumped through tubes on the side of the human-on-chip plate and routed from tube to tube via a valve system.

[0120] In one embodiment, the individual biochip entities are assembled as a single element within a housing and valve mechanism and connected via a cylindrical rotating mechanism, with fluids passing through tubes on the side of the human-on-a-chip plate and pumping from tube to tube via the valve system.

[0121] In one embodiment, the individual biochip entities are incorporated into a human-on-a-chip plate where a cylindrical rotating mechanism connects the housing and a valve mechanism connecting the individual chip components, and fluids are extracted from specific regions within each biochip in the human-on-a-chip plate.

[0122] In additional embodiments, the human-on-a-chip plate can have biochips, individually removable biochips. The individually removable biochips are assembled in a puzzle-like fashion by interconnecting the different chips via tubes to the inlet / outlet valves on the biochip and valve-holding plate. Or, the individual biochip entities are incorporated as one human-on-a-chip plate, where a cylindrical rotating mechanism connects the housing and the valve mechanism connecting the individual chip elements. In one embodiment, two tube chips can be connected to one tube chip.

[0123] 14A shows a schematic diagram including pressure sensors at the inlet and outlet fluid ports of each chip to manipulate the pressure within the channels of the chip. In an additional embodiment, thanks to the pressure sensors of each chip, fluids are controllably perfused from the tubular channels to the interstitial channels or vice versa.

[0124] In additional embodiments, the biological system-on-a-chip plate includes a plurality of tubular biochips, a fluid channeling means in fluid communication with at least one of the biochips. The fluid channeling means includes a plurality of fluid paths and a selectable connection means providing the ability to selectively connect or disconnect the fluid communication. In additional embodiments, the tubular biochip has an inlet port and an outlet port. In one embodiment, at least two biochip ports are fluidly connected to at least one fluid path. In one embodiment, the fluid channeling means allows for the transport or circulation of media through the tubular or interstitial regions of the plurality of biochips. The selectable connection means allows for serial or parallel arrangement of the biochips depending on the particular experiment or application. In additional embodiments, at least one selectable connection means is configured to allow for a permanent selection process to connect (e.g., a rupturable frangible seal) or disconnect (e.g., a heat sealable thermo-adhesive material) the fluid communication therethrough. In one embodiment, at least one selectable connection means includes a valve means providing reversible connection and disconnection. Valve means allow isolation of the biochip during an experiment.

[0125] In one embodiment, the valve means allows access to remove media samples during or after an experiment. The valve means allows for media to be introduced before or during an experiment. Furthermore, the valve means allows for control over where different media, particularly cell types, are placed during an experimental setup where media is introduced before or during an experiment. In one embodiment, the valve means is stable in both connected and disconnected states. Additionally, several other elements can be utilized, such as pinch valves with cams (friction hold, or over-center profile), rotary valves (friction cylinder or ball valves), pinch valves with snap-through bi-stable levers, self-stabilizing pilot pneumatic valves, etc.

[0126] In additional embodiments, the biological system-on-chip plate includes a number of valve means actuated by a number of actuators, where the number of actuators required to actuate the valve means is less than the number of valve means. In one embodiment, the biochip is a multi-tube chip. In one embodiment, the biochip is an organ-on-chip organism simulation. In additional embodiments, a first biochip mimicking a first organ type is connected to a second biochip mimicking a second organ type. In one embodiment, the biological system-on-chip plate mimics multiple organs of an organism. The biological system-on-chip plate mimics enough organs to gain insight into the effects of experimental compounds on multiple organs within the organism. In one embodiment, the organism is a human. In additional embodiments, the biological system-on-chip plate is configured to engage with external actuation and pump means, allowing the biological system-on-chip plate to be low cost or disposable replaceable.

[0127] MICRO-VALVING MECHANISM

[0128] Figures 15A-15C show perspective views of various types of mechanical microvalves. In one embodiment of the mechanical microvalve of Figure 15A, two pistons move in opposite directions to open and close two ports that are placed in close proximity and are required to always have opposite (on-off) configurations. The hydraulic pistons are connected to a volume of incompressible fluid, so that as one piston moves in one direction, the fluid acts as a push or pull on the other piston, moving in the opposite direction.

[0129] Figure 15B shows a perspective view of a rotating cylinder mechanical microvalve, in which a drill cylinder containing inlet and outlet ports rotates around an axis to align holes to the desired locations and connect fluids to microfluidic channels outside the system for fluids entering the chip.

[0130] FIG. 15C shows a perspective view of a pneumatically actuated drilled piston microvalve, in which a drilled piston connecting the ends of a microfluidic channel is pneumatically actuated to move the piston and block the channel.

[0131] Figures 16A-16F show several embodiments of an electrically actuated solenoid bistable microvalve. This microvalve includes a beam with a metal end between two magnets, where an electrical signal triggers the direction of the magnet's force to move the beam between two locations. On the other side of the beam is a hole that opens the channel when it aligns with the microfluidic port opening of a connected microchannel, and closes the channel when it is not aligned.

[0132] Figures 16A and 16C show two embodiments of an electrically actuated solenoid bistable microvalve where both drilled beams are in the extended position, thus causing the microvalve to be in the open position.

[0133] Figures 16B and 16D show two embodiments of an electrically actuated solenoid bistable microvalve, where the internal elements of the microvalve are shown with one beam in its minimum extended position, blocking the microfluidic channel, and the other beam in its maximum position, with a hole drilled in it aligned with the hole in the microfluidic channel it acts on, opening the flow in the microfluidic channel.

[0134] Figures 16E and 16F show the assembly of an electrically actuated solenoid bistable microvalve on a human-on-a-chip plate, where the plate in Figure 16E shows the microfluidic channeling that the microvalve is trying to block or open. Furthermore, Figure 16F shows a top view of the human-on-a-chip plate with the microvalve and its electrical circuitry.

[0135] Figures 17A-17D show various embodiments of pneumatically actuated drilled piston microvalves: Figure 17A shows a perspective view of a normally closed microvalve in which the drilled piston holes are not aligned with the holes that connect to the microfluidic channels.

[0136] FIG. 17B shows an exploded view of a normally-closed microvalve in which the drilled piston holes are not aligned with the holes that connect to the microfluidic channels.

[0137] FIG. 17C shows a perspective view of a normally open microvalve with the drilled piston holes aligned with the holes that connect to the microfluidic channels.

[0138] FIG. 17D shows an exploded view of a normally open microvalve with the drilled piston holes aligned with the holes that connect to the microfluidic channels.

[0139] 18A shows a perspective view of a bistable beam on a flexible tubing microvalve connecting individual biochip elements of a human-on-a-chip plate according to one example of the present disclosure. In one embodiment, the valve mechanism connecting the individual biochip elements is bistable, with the microvalve having two stable positions. The bottom balloon channel is pneumatically actuated, and when air flows through the channel, all channels in the microvalve open, while the top set of balloons are individually pneumatically actuated, each closing the channel below.

[0140] FIG. 18B shows an exploded view of a bistable beam on a flexible tubing microvalve connecting individual biochip elements of a human-on-a-chip plate according to one example of the present disclosure. In one embodiment, the valve mechanism connecting the individual biochip elements is bistable, meaning the microvalve has two stable positions. Power is only required to switch between the two states, and when actuated, the bistable beam remains in one of the two bistable positions. The bottom balloon channel is pneumatically actuated, opening all channels of the microvalve when air flows through the channels, while the top set of balloons are individually pneumatically actuated, each closing the channel below it by squashing the tubing to block flow.

[0141] 19A shows a perspective view of a bistable magnetic drill piston microvalve connecting individual biochip elements of a human-on-a-chip plate according to one example of the present disclosure. In one embodiment, the valve mechanism connecting the individual biochip elements is bistable, and the microvalve has two stable positions. The lower and upper channels are magnetic, making the microvalve bistable, and the drill piston is pneumatically actuated to control the state of the microvalve.

[0142] FIG. 19B shows an exploded view of a bistable magnetic drill piston microvalve connecting individual biochip elements of a human-on-chip plate according to an example of the present disclosure. In one embodiment, the valve mechanism connecting individual biochip elements is bistable, and the microvalve has two stable positions. The lower and upper channels are magnetic, and the drill piston is attached to the surface of the channel to make the microvalve bistable. The drill piston can be replaced by a ball microvalve. The drill piston is pneumatically actuated to control the open and closed states of the microvalve. The drill piston can be any piston with a hole, which in the open position is concentric with the duct, allowing fluid to pass through.

[0143] 20A, 20B, 20C, and 20D show different perspective views of a bistable mechanically actuated flexible tube microvalve connecting individual biochip elements of a human-on-a-chip plate according to an example of the present disclosure. In one embodiment, the valve mechanism connecting the individual biochip elements is bistable, and the microvalve has two stable positions. The bistable beam is mechanically actuated, where when closed, it crushes the flexible tube to block fluid from passing through. This design can be solenoid or pneumatically actuated to move a main beam that actuates the bistable beam, and the bistable beam can be replaced by a bistable magnetic piston.

[0144] 21A, 21B, 21C, and 21D show different perspective views of a mechanically actuated pinch ball open / close microvalve connecting individual biochip elements of a human-on-a-chip plate according to an example of the present disclosure. In one embodiment, the valve mechanism connecting the individual biochip elements can be either open or closed, rotating a microvalve with a ball with a slotted hole to allow fluid to pass or block fluid. The ball microvalve is mechanically actuated by a motor controlled by a robotically actuable system that moves in 2D or 3D planes to open and close the microvalve, or actuated by a stand-alone system that can pre-configure the configuration of the microvalve to mimic organs in the human body.

[0145] 22A, 22B, 22C, and 22D show different perspective views of permanently sealed open / closed microvalves connecting individual biochip elements of a human-on-a-chip plate according to one example of the present disclosure. In one embodiment, the valve mechanism connecting the individual biochip elements is either open or closed, the permanent seal is preset once to have either an open or closed microvalve, and a pin is permanently inserted into the channel, which either blocks or opens the channel depending on the shape of the pin.

[0146] OPERATING SYSTEM COMPONENT ASSEMBLY

[0147] 23A-23K show perspective views of one embodiment of an assembled operating system. In one embodiment, the operating system is comprised of multiple interconnected subsystems: a mixing chamber, a fluid control operating system, a human-on-a-chip plate, individual biochip elements, a machine control system, and an analyzer system.

[0148] Figure 23G shows ports on one side of the device connecting the operating system with tubing exiting the chip to the waste chamber. It also shows a port connecting the operating system to a pneumatic air signal inlet that actuates a valve that controls flow to the waste chamber. In addition to fluid tubing that pumps temperature controlled liquids through the system to heat it, there is a pneumatic port that actuates a valve that controls the flow of thermally controlled fluids to distribute heat evenly throughout the system.

[0149] Figure 23F also shows two ports on the back of the device that connect to a pneumatic control device. One port contains 11 tube fittings to pneumatically control the 33 valves in the chip, and the other port contains 16 pneumatic tube fittings to control the 72 valves of the 72 media microchambers that feed into the 72 different channels of the human-on-a-chip plate. On the same side, the device has 72 fluid ports that connect fluids coming from outside the system to the media microchambers. These fluids flow from the mixing chamber to a valve distributor and then to these ports.

[0150] Figure 23G shows the temperature, pressure / flow, and O 2 and CO 2 Shown is the side with the four electrical plugs that each control a sensor.

[0151] Figure 23H shows the human-on-a-chip plate connected to the fluidic operating system, and Figure 23I shows the ports of the fluidic operating system device that connect the biological fluid ports to each channel of the human-on-a-chip plate, the pneumatic control ports that connect to the valves in the human-on-a-chip plate, and the heating fluid ports that connect and heat the human-on-a-chip. Figure 23J shows the three main elements in the system: the fluidic operating system device, the media chambers that connect to it, and the human-on-a-chip plate.

[0152] In one embodiment, a fluid (herein referred to as media) consisting of cells or biological or chemical components is first manually inserted into the mixing chamber by a user. After entering the mixing chamber, the media is pumped through a pressure sensor, a bubble filter, and a 1:12-way valve into the tubing of the fluid operating system.

[0153] The 1-to-12-way valve allows the medium to pass from the predefined mixing chamber through the operating system and then be distributed by the valve system, which is the first subsystem of the fluid operating system. The distribution valve is composed of a combination of valves, where one 1-12-way valve is connected to twelve other 1-6-way valves to form a 1-72 routing path. The medium then reaches the microchambers in the immediate vicinity of the human-on-chip. The microchambers are interacted with a pumping mechanism, and the fluids in the microchambers are precisely pumped to the channels or waste chambers in the chip entity of the human-on-chip.

[0154] In one embodiment, the microfluidic pumping mechanism controls the precise amount of fluid pumped into the microchamber and from the microchamber to the chip, which can be as small as 0.1 pl.

[0155] In one embodiment, each channel of the chip has an inlet and an outlet port, and a pump control mechanism can control the pressure, flow rate, and direction of flow.

[0156] In one embodiment, the at least one analytical device may be used for real-time measurement of a chemical, biological or physical parameter.

[0157] In one embodiment, the operating system provides pressure sensors at each inlet and outlet fluid port of the chip to manipulate the pressure within the channels of the chip, thereby allowing fluid to be controllably perfused from the tubular channels to the interstitial channels and vice versa.

[0158] In one embodiment of the operating system, at least one valve system may be capable of opening, closing and altering the flow paths to all compartments while an experiment is running.

[0159] In one embodiment of the operating system, multiple organ-on-a-chip tube scaffolds on a human-on-a-chip plate are connected to each other to allow continuous flow.

[0160] In one embodiment of the operating system, multiple organ-on-chip tube scaffolds on a human-on-a-chip plate are connected to each other for a controllable amount of time.

[0161] In one embodiment of the operating system, multiple organs-on-chips on a human-on-chip plate are isolated from each other.

[0162] In one embodiment of the operating system, a microfluidic automated flow control mechanism allows for the addition and removal of fluids in the organ-on-chip tubular scaffold.

[0163] In one embodiment of the operating system, a microfluidic automated flow control mechanism enables the addition and removal of fluids in multiple organ-on-chip tube scaffolds on the human-on-chip plate.

[0164] In one embodiment of the operating system, a microfluidic automated flow control mechanism allows for the addition and removal of fluids to the external compartment surrounding the organ-on-chip tubular scaffold.

[0165] In one embodiment of the operating system, a pump control mechanism can transfer pumped fluid to a waste chamber.

[0166] In one embodiment of the operating system, a pump control mechanism can transfer the pumped fluid to a sample chamber.

[0167] In one embodiment of the operating system, the pump control mechanism can transfer the pumped fluids to different biochips.

[0168] In one embodiment of the operating system, a pump control mechanism can transport the pumped fluid for extraction.

[0169] In one embodiment of the operating system, an electro-mechanically controlled robotic arm embedded in and interconnected with the operating system can move in at least one dimension to perform different movements.

[0170] In one embodiment of the operating system, an electro-mechanically controlled robotic arm embedded and interconnected with the operating system can place at least an embedded microscope, spectrometer or surveillance sensor on top of the human-on-a-chip.

[0171] In one embodiment of the operating system, an electromechanically controlled robotic arm embedded and interconnected with the operating system can have at least one built-in pipette capable of withdrawing a sample from at least one compartment of the sample source chamber.

[0172] In one embodiment of the operating system, an electro-mechanically controlled robotic arm embedded and interconnected with the operating system can have at least one socket head capable of engaging and actuating at least one type of mechanically actuated valve.

[0173] OPERATING SYSTEM FLUIDIC CONTROL MECHANISM

[0174] In one embodiment, as shown in Figures 23F, 23G, and 23H, the fluid control mechanism allows the use and manipulation of microliters of fluid used in biochip cell culture without losing milliliters of fluid due to volume loss in tubing reaching the biochip channels. The biochip channels, valve mechanisms, and close proximity to small individual media chambers for each inlet and outlet hole in the system play a key role in this system. In one embodiment, a piston pump mechanism functions to insert fluid into the individual media chambers and pump it out to the biochip.

[0175] Figures 25, 26, 27 and 28 show some designs of the fluid control mechanism of the operating system based on the interaction of the channels of the biochip with the channels of the machine. The machine controls the movement of the valves, mainly the inlet / outlet port valves, built into the biochip with a mechanism that depends on the type of valve. In one embodiment, if the valves are mechanical, they can be actuated via a micro linear actuator within the system. Furthermore, if the valves are normally open / closed valves, they can be actuated mechanically or pneumatically via an actuator built into the system or via an actuator interface within the system that is actuated from outside the system.

[0176] Figure 28 shows the interface of the biochip with the operating system. The first level of fluid control beyond opening and closing the inlet / outlet holes and the separate compartment interfaces are three series valves. The first valve is an on-off valve that opens and closes the biochip channel to the operating system. The second valve is a waste channel redirection valve that switches the flow direction to either the second on-off valve or the waste microchannel. The waste microchannel is integrated into the human-on-chip if this valve is on the biochip, or onto a layered plate that interacts with the biochip if this valve is in a plate rather. The third valve is a second on-off valve that uses the operating system to separate the waste valve and the first on-off valve that connects to the biochip channel from the medium chamber plate. The three series valves and the waste microchannel can be integrated into the biochip itself if the valves used are inexpensive, as is the case with membrane deformation valves that are pneumatically actuated from outside the system. Alternatively, the operating system valves can be integrated into a separate plate assembled on top of the biochip, primarily if the valves are expensive or are actuated from the same location as the valves, such as in the case of solenoid magnet actuators or squiggle motor actuators.

[0177] At the interface with the chip holes, a plate can be added that can form different routings for different tubes and microfluidic channels. This chip plate can be the basis when valves are not included in the human-on-chip design or when this routing plate connects different individual chips to create new human-on-chip assemblies.

[0178] At the interface of the three serial valves, the media chamber plate of the individual holes is interacted. The operating system chamber can be integrated on the surface of the biochip if the three serial valves are also integrated in the biochip, but in preferred circumstances can also be integrated in a separate plate that interacts with the surface of the biochip or the serial valve plate surface. Each individual media chamber in this plate is preferably fed by a single inlet / outlet hole, accommodates several hundred microliters of fluid, and is actuated from above by a piston integrated in the plate.

[0179] At the interface with the media chamber plate is a piston plate that contains pistons that actuate individually for each media chamber. Each piston of the piston plate is composed of two sub-parts. The lower part of the piston has a cross-section at the bottom that is the same size and shape as the media chamber it operates. In addition, the piston contains a channel over its entire length that leads to the upper part of the piston. The upper part of the piston contains an interface with the lower piston part on its lower surface. This interface opens into a channel that connects the channel of the lower piston part that is open at the media chamber to the side of the upper part of the piston where the tube interface that carries the medium from the media chamber through the piston towards the upper valve plate is located. The complete piston plate is assembled by inserting the lower piston part from below into the plate that contains the holes for the pistons and the upper piston part from the opposite side into the respective piston. The upper surface of the upper piston part contains a pressure plug interface feature that interacts with the respective piston actuator contained in the piston actuator plate.

[0180] Interfacing with the piston plate is the piston actuator plate, which contains an actuator for each piston. The actuators can be connected directly to the pistons or through a connector that reaches the actuators outside the system. As shown in Figures 27I and 27J, the actuators included in the operating system that interact with the piston plate are small in size, have minimal heat dissipation, and are better suited to move small, controllable steps, a perfect example of which is the Squiggle motor. Other motors such as solenoid magnetic actuators or small stepper motors can also be used in the system. For the actuators outside the operating system, any type of actuator can be used as long as the interface errors are minimized. Thus, pneumatic or hydraulic actuators can be used to actuate the pistons from outside the system, as long as the tubes connecting the external actuators to the system are thermally controlled sealed and the compressibility of gases is taken into account.

[0181] The tubes connected to the pistons feeding the small media chambers pass through an upper on-off valve plate at a minimal distance from the piston plate above or to the side. The on-off valve plate contains valves connecting and isolating each tube from the small media chamber through the piston to the media source plate. The individual on-off valves can be any valve of small capacity or size, on-spot actuated solenoid magnetically actuated valves, or other valve types that are electronically actuated from their location. They can also be membrane or piston based valves that are pneumatically or hydraulically actuated via tubes that reach actuators external to the system.

[0182] The tubing coming out of the top valve plate is fed to the fluid source plate. The fluid source plate is the plate to which the mechanism for fluid insertion is connected. Individual syringes fit into the inlets of the fluid insertion plate which can be used for several types of experiments. Also, a media source chamber plate mechanism can be connected to the media source plate. This media source chamber plate has its own valve and pump system which is external to the operating system but can be controlled by the same software. The main external chamber is connected to a valve plate with a mesh of microvalves to open, mix and flow media from any media chamber in the external plate to any inlet of the source plate of the operating system. From the other side of the fluid source plate in the operating system, a three-way valve can be built into the system which acts as a pressure relief outlet and also removes air bubbles from the system that may arise from the interface while connecting the tubing or syringes to the operating system.

[0183] Fluids, cells, and other biological components exiting the system can be pumped out and extracted and dispensed at any designed time interval by opening and closing the respective valves. The tubing exiting the system is also monitored by a Fourier transform infrared spectroscopy (FTIR) device, O 2 and CO 2 It is interconnected with various measurement technologies such as sensing devices, pH sensing devices, etc.

[0184] In one embodiment, the human-on-a-chip plate includes a manifold that allows the internal space of the system to be maintained at specific environmental conditions that can be tailored to a particular test protocol. Additionally, heat dissipation or generation devices can be used to mitigate undesirable temperature conditions.

[0185] In one embodiment, the human-on-a-chip plate includes a fluidic control system that can deliver and / or collect the reagents required to perform analytical assays at precise times and channels of one or more biochips within the operating system, as shown in Figures 29A and 29B. These assays may be measured using built-in microscopes, spectrometers and / or FTIR, or other suitable analytical devices, whether built-in or not.

[0186] In one embodiment, the human-on-a-chip plate includes a fluid control system that can deliver and / or collect liquid samples from any channel of any biochip within the operating system. The liquid samples collected or delivered may contain any desired chemical or biological material, including but not limited to cells, enzymes, drugs, growth factors, nutrients, etc. The amount of sample delivered to each channel can be between 0.5-25 pl.

[0187] In one embodiment, the human-on-a-chip plate includes a fluid control system that can pump liquid samples from any channel of any biochip in the operating system. These can be pumped from an inlet hole on the top or from the side. The fluids pumped to multiple channels in each biochip and / or across different biochips in the operating system can be the same or different. Gases can also be pumped instead of fluids to one or more channels. Liquids or gases can be pumped at different flow rates. The pumped liquids can include chemicals, drugs, polymers, biological materials, cells, growth factors, nutrients, or other substances.

[0188] In one embodiment, the human-on-a-chip plate includes a fluid control system capable of delivering a polymerizable material, which may contain cells or other biological or chemical substances, to any channel or channels (interstitial and / or tubular) of one or more biochips within the operating system. This control system is coupled to a temperature controller such that the temperature is maintained such that the polymerizable material is in a liquid state until it is delivered to the desired tubular. Once delivered, the temperature is increased to ensure gelation.

[0189] 30A and 30B show flow diagrams for delivering the sample from the media chamber to the biochip and from the biochip to a measurement device such as an ATR FTIR, respectively.

[0190] MIXING CHAMBERS

[0191] 32A, 32B, 32C, and 32D show schematic diagrams of a mixing chamber and some of its components, according to one example of the present disclosure. In one embodiment, a fluid (herein referred to as media) consisting of cells or biological or chemical components is first manually inserted by a user into the mixing chamber, where gas is bubbled and a heating element heats the fluid. The system then automatically distributes the media to one or more biochips, waste chambers, or other elements of the operating system. 2 , CO 2 and temperature sensors are used to maintain physical parameters at levels set by the user.

[0192] In one embodiment, the pump mechanism is pressure driven and is pneumatically actuated via multiplexed air tubing from a pressure control system.

[0193] In another embodiment, the pump system mechanism is driven by a syringe pump, with fluid being forced through the system via a syringe.

[0194] In another embodiment, the pump mechanism is a peristaltic pump drive, whereby fluid within the container is drawn into the system.

[0195] 33 shows several example flow diagrams of embodiments of a mixing chamber in one example of the disclosure. In one embodiment, after the media enters the mixing chamber, it is pumped into a fluid control operating system and while in the mixing chamber, the fluid passes through a pressure sensor, a bubble filter, and a 1 to 12 way valve that takes the fluid from one of the mixing chamber's reservoirs into tubing of the fluid operating system.

[0196] CO 2 , O 2 SENSING MECHANISM AND CONTROL FOR THE CO 2 , O 2 AND TEMPERATURE)

[0197] FIG. 34A shows an example of the present disclosure in which all containers are O 2 , CO 2 and N 2 Bottles and O 2 and CO 2 FIG. 1 shows a flow diagram of one embodiment of a mixing chamber with access to a sensor. While the medium is in the mixing chamber vessel, gas is bubbled in and a heating element heats the fluid. 2 , CO 2 and temperature sensors are used to maintain physical parameters at levels set by the user.

[0198] In one embodiment, O 2 , CO 2 The temperature control system is applied and measured not only in the mixing chamber but also at the level of the cells and media within the chip entity, as shown in Figure 34B. This provides insight into the metabolic activity of the cells and allows for more precise control of their physical parameters.

[0199] In one embodiment, O 2 and CO 2is pumped directly into the mixing chamber via a closed-loop control system, and optical or biochemical sensors measure the amount of O diffused through the fluid. 2 and CO 2 Measure the ratio.

[0200] In one embodiment, the temperature is induced by a heating element and measured using a thermal sensor, and the temperature is controlled by a feedback control system as shown in Figures 35A and 35B.

[0201] SOLUTION INSERTION

[0202] In one embodiment, as shown in FIG. 28B, to fill the media chamber, the user of the system performs two steps repeatedly until the media chamber is filled. In the first step, the valve below the media chamber is closed to block the flow to the biochip or waste channel, and the valve above the media chamber in front of the media source plate is opened to introduce the injection from the source plate. After the valve is opened, the piston is raised to pump the fluid from the source plate towards the media chamber. Because the volume of the media chamber is small (a few hundred microliters), the volume of the tubing that reaches the media chamber from the source plate is large, and therefore the piston reaches its top position before the media reaches the media chamber in the tubing. Once the piston reaches its top position, the valve below the media chamber and the waste channel valve are opened, and the valves towards the biochip channels and the top valve in front of the media source plate are closed. After opening and closing each valve, the piston is pushed down to remove excess air from the media chamber. Once the piston reaches its bottom position, steps 1 and 2 are repeated until the media fills the tubing and reaches and fills the media chamber. In one embodiment, the solution can be a cell culture media.

[0203] SOLUTION REPLACEMENT

[0204] As shown in FIG. 28C, to empty the media chamber, the user of the system repeats two steps until the media chamber is empty. In the first step, the valve below the media chamber is closed to block flow to the biochip or waste channel, and the valve above the media chamber in front of the media source plate is opened to push the fluid into the source plate. After the valve is opened, the piston is pushed down to pump the fluid from the media chamber towards the source plate. Because the volume of the media chamber is small (a few hundred microliters), the volume of the tubing that reaches the media chamber from the source plate is large, so the piston reaches its bottom position before the media reaches the source plate in the tubing. Once the piston reaches its bottom position, the valve below the media chamber and the waste channel valve are opened, and the valves to the biochip channels and the top valve in front of the media source plate are closed. After opening and closing the respective valves, the piston is pulled up to fill the media chamber with additional air. Once the piston reaches its top position, steps 1 and 2 are repeated until the media empties the tubing and reaches the source plate. In one embodiment, the solution can be a cell culture medium.

[0205] CELL INSERTION

[0206] To seed cells into the biochip channels described in FIG. 28D, the user first inserts the media containing cells into the media chamber and fills the media chamber in the same manner as described in FIG. 28B. The user then opens the valves below the media chamber on either side of the waste valve to allow access to flow to the biochip and closes the valves of the waste channel to block flow to the waste channel. The user then closes the valve above the media chamber in front of the media source plate to block flow to and from the source plate. After opening the valves, the user opens the outlet waste valve to push any excess air or media out of the system and replace them with the cell-loaded media, then pushes down the piston to pump the fluid from the media chamber towards the biochip channels. The user can optionally open channels between different biochips from the outlet to seed the same cells onto different biochips, so that the open outlet waste channel is the last biochip the cells need to reach. Once the cells reach the desired location, close all valves to stop pumping and wait for the cells to attach. Once the cells have settled and attached to the bottom of the tube, flip the whole operating system, open the same valves and perform the same procedure, pumping the cells into the biochip tube and waiting until the cells have attached to the top of the tube. Once the cells have settled and attached to the top of the tube, flip the whole operating system back to the initial shape and continue the experiment.

[0207] In one embodiment, cells can be inserted into the biochip by any available channel, including but not limited to a side channel. The cells can be inserted suspended in a suitable liquid or a suitable polymer that will solidify.

[0208] In one embodiment, skin tissue is cultured onto multiple biochips to enable the human-on-a-chip operating system to execute protocols related to, but not limited to, assessment of skin health and physiology.

[0209] In one embodiment, cells, media, or other materials may be inserted into any channel of the biochip or into any biochip in an operating system at a specific location. Cells may also be extracted by, but not limited to, delivering an appropriate enzyme or other reagent that causes detachment of the cells at the specific location. Once the cells are detached, the fluidic control system collects the suspended cells.

[0210] Media extraction (MEDIA SAMPLING)

[0211] In one embodiment, to collect a media sample from the biochip channel described in FIG. 28E, the user of the system performs two steps repeatedly until the media chamber is filled with the intended sample volume. In the first step, for both the inlet and outlet fluid channels, the valves below the media chamber on either side of the waste valve are opened to allow access to the flow from and to the biochip, and the valves of the waste channels are closed to block the flow to the waste channels. Next, the valve above the media chamber in front of the media source plate is closed to block the flow from and to the source plate. After opening and closing the intended valves, the piston controlling the inlet fluid is pushed down and the piston controlling the outlet piston is pulled up at the same speed and volume to collect the intended sample volume in the outlet media chamber. After collecting the intended sample in the media chamber, the outlet media chamber is emptied to initiate step 2, which transfers the collected sample to the top of the system. The emptying process is as previously described in FIG. 28C.

[0212] In one embodiment, media samples are collected from any biochip channel within the operating system. In some cases, media or other liquid material is collected from a side channel.

[0213] Cell extraction (CELL SAMPLING)

[0214] In one embodiment of the operating system, the same cell or tissue type can be grown across different chips with a common tubing, so that each biochip represents a subsample of the same organ. Each individual biochip can then be independently removed from the circulation system at different time points and taken out of the system for downstream analysis, allowing for extractions at different time points.

[0215] In one embodiment of the operating system, the same cell or tissue type can be grown across different biochips with a common tubing. Upon drug administration, the biochips can be separated from each other. Each individual biochip can then be individually extracted or removed for downstream analysis.

[0216] To collect a cell sample from the biochip channel described in Figure 28F, four steps are performed until the media chamber is filled with the intended sample volume. In the first step, the biochips are connected along the tubes to make one continuous tissue sample, so that each biochip collected at different time lapses is a sample of the same tissue. Therefore, this interface is necessary in the first step. In the second step, the valves between the different biochip entities are closed to make each biochip tube a sample to be collected at a specific time. In this stage, the media in the inlet media chamber of the biochip where the cell sample is collected is replaced with trypsin by blocking the destination valves and pumps, as described above. In the third step, the sample of cells is collected in the outlet media chamber in the same way as how the media sample is collected by pumping trypsin into the destination media channel, as described above. In step 4, after the desired sample is collected in the media chamber, the outlet media chamber is emptied to move the collected sample to the top of the system. The emptying process is as described above in Figure 28C.

[0217] In one embodiment, cells, media, or other materials can be extracted from specific locations within any channel of the biochip or from any biochip within the operating system. Cells can also be extracted by, but not limited to, delivering appropriate enzymes or other reagents that cause cell detachment at specific locations. Once the cells are detached, the fluidic control system collects the suspended cells. Extraction can occur from inlet and outlet holes of the biochip. In some cases, cell extraction can occur from side channels of the operating system.

[0218] INSERTING MULTIPLE REAGENTS SEQUENTIALLY

[0219] In one embodiment of the operating system, the reagents can be added sequentially. The media chamber is filled with a first reagent as described above, which is inserted into the interstitial chamber of the biochip. The media chamber is then emptied and filled with another reagent as described above, which is inserted into the interstitial chamber. This can be repeated as many times as necessary. The time intervals between different reagents being added to the interstitial chamber are controllable and programmable. The reagents can be added immediately or one after the other after a set time interval.

[0220] In one embodiment, sequential insertion of reagents is essential not only when administering multiple chemicals, but also when performing assays including, but not limited to, chip immunohistochemistry and cell health assays.

[0221] TUBULAR ARCHITECTURE AND BARRIER FUNCTIONS OF DUCTAL TISSUES

[0222] Figure 39E shows an embodiment of the system used as a blood-brain barrier model for drug screening and penetration studies, where a tube is loaded with brain microvascular endothelial central nervous system cells interacted with tissue composed of pericytes, astrocytes and neurons, which can be used to perform drug permeability studies and / or toxicity and efficacy analysis on neural tissue.

[0223] CULTURING OF MULTIPLE TISSUE IN DIFFERENT CHIPS CONNECTED BY A COMMON DUCT / VESSEL

[0224] Figure 39B shows a perspective view of an embodiment of multiple cultures in our chip connected to a common tube / vessel. In one embodiment of the operating system, it can be used to study the microbiota-gut-brain axis. Here, the interstitial chamber of one biochip contains intestinal epithelial cells overlaid with intestinal mucus and patient-derived microbiota. The medium in the interstitial chamber is hypoxic. The tubes connecting the various biochips are seeded with vascular endothelial cells and the medium flowing through them is fully oxygenated. The other biochip contains cultured neurons, astrocytes and microglia embedded in a hydrogel matrix. The impact of the different microbiota on the brain biochip can be evaluated by microscopy or by extracting the neuronal cells and performing ex situ protein and gene expression assays.

[0225] FIG. 39B is a perspective view of an embodiment of multiple cultured tissues in our chip connected to a common tube / vessel. In an embodiment of the system, each biochip contains different tissues and organ systems, which can be configured to be connected in parallel and / or series by a common tube, or tubes that mimic the vascular and / or lymphatic systems. Some biochips can have multiple tubes that can be interconnected with one or more of the other biochips. Taken together, this becomes a human-on-a-chip, which is useful for testing off-target and / or systemic effects of drugs, and the permeability of drugs through different anatomical barriers. It can also provide a more accurate indication of the in vivo half-life of drugs.

[0226] In one embodiment, the system may be configured to simultaneously evaluate liver and kidney toxicity in drug screening. As both kidney and liver are responsible for xenobiotic removal and metabolism, it is essential to evaluate the effectiveness of a drug in the presence of these two tissues. In this embodiment, one or more biochips contain renal epithelial cells and another (or multiple) biochip liver cells, as described elsewhere herein. The two tissues are connected by a tube lined with endothelial cells. The drug dissolves in the tube and cytotoxicity, proliferation and apoptosis assays can be performed on the two biochips using spectroscopic assays. The system may also be configured to include one or more tissues that are target organs for the tested drug on separate biochips.

[0227] Figure 39A shows a cross-sectional view of an embodiment of our two-tube chip tissue tubular structure. In one embodiment, the system allows for the growth of at least two different tissues in direct contact (one in a tube surrounded by a second tissue) or in contact via one or more vessels (blood vessels or lymphatic vessels or both). This allows for the study of tissue interactions.

[0228] In one embodiment of the system, the biochip plate can contain different independent biochips, each with a specific tissue, and simultaneously contain a set of different biochips interconnected with the other chips by one or more tubes to form a human-on-a-chip.

[0229] LIQUID-AIR-INTERFACE LUNG-ON-A-CHIP

[0230] As shown in FIG. 39C, in one embodiment, the system can be used in a model for pulmonary gas exchange testing to study lung physiology, disease, and the effects of drug candidates. Human epithelial and pulmonary endothelial cells are cultured within the interstitial gel in an external compartment that contains one or two tubes. One tube has human small airway epithelial cells attached and grown on the inner wall of the tube scaffold. Air is pumped through the lumen. If a second tube is present, the second tube may contain vascular cells, epithelial cells, and pericytes to replicate capillaries. This tube has a constant flow of red blood cells at 5-0.03 generations and a constant pressure of 30 mmHg. Similarly, in one embodiment, air is pumped through the interstitial channels of the chip and cells can be grown in one or more tubes present in the chip.

[0231] IMMUNE SYSTEM MODEL

[0232] As shown in Figure 39D, in one embodiment, the system allows for elements of the circulating immune system, such as, but not limited to, T cells, natural killer cells, and / or B cells, to be added to an individual chip or multiple chips within the system, allowing for the study of tissue (or tumor)-immune system interactions as well as the study of the efficacy of immunotherapies.

[0233] In one embodiment, the system allows for the addition of engineered immune elements, such as, but not limited to, CAR-T cells, to an individual chip or multiple chips within the system, allowing the system to be used to assess efficacy and toxicity of immunotherapies.

[0234] One embodiment of the operating system involves modeling of the immune system, including one or more of the following organs mimicked on one or more chips in the system: spleen, bone marrow and / or lymph nodes, which can be used as standalone organs or connected to other organs to study interactions between the immune system and other components.

[0235] BIOPSY CULTURE FOR PRECISION & PERSONALIZED MEDICINE DRUG PROGNOSIS

[0236] In one embodiment, the system may be used for precision medicine, where patient biopsies or primary cells can be cultured in 3D space to recreate physiological / disease structures. Drugs can then be administered to the system according to the processes and methods disclosed herein, and specific toxicity and efficacy tests can be performed to quantify the efficacy of the drug or drug combination.

[0237] In one embodiment, primary cells from different patients can be grown on different biochips independent of each other.

[0238] In one embodiment, biochips containing different cells of the same patient can be connected to each other to form a person-on-plate.

[0239] Microscopy Measurement Interface

[0240] In another embodiment of the operating system, the system can perform real-time live cell imaging and end-point imaging for visualization of, but not limited to, cells, cellular components, and / or tissue morphology, which may be useful for assessing cell proliferation, migration, and / or death, among other things. The integrated system software then analyzes the images to provide quantitative data.

[0241] In one embodiment of the operating system, tissue culture imaging is performed using fluorescence and brightfield microscopy over time. In additional embodiments of the operating system, the system can determine and complete all procedures to perform immunohistochemistry and / or immunofluorescence staining, followed by automated imaging using fluorescence and / or brightfield microscopy, followed by high throughput image analysis.

[0242] In another embodiment of the operating system, the operating system may be used for drug and pharmaceutical ingredient testing and screening.

[0243] SPECTROSCOPY MEASUREMENT INTERFACE

[0244] In one embodiment of the operating system, high throughput absorbance and colorimetric assays can be performed to assess the physiological state of the tissue. Some, but not all, specific parameters that can be measured include cell health, viability, enzyme function, or cytochrome activity. In another embodiment of the operating system, high throughput fluorometric assays can be used to assess cell metabolism, viability, health, cell death, ADME toxicity, or other parameters. In another embodiment of the operating system, high throughput luminescence assays can be performed to assess cell health, metabolism, viability, cell proliferation, ADME toxicity, luciferase activity, inflammation, oxidative stress, or apoptosis, and / or other assays using this technology.

[0245] In another embodiment of the operating system, a combination of fluorescence, colorimetric, absorbance and emission spectroscopy allows single or multiplex assays to assess cellular functions and processes, including but not limited to basic indicators of cell health (cell proliferation, cell viability and cytotoxicity, specific mechanisms of cell death such as apoptosis or necrosis) and / or cellular processes (e.g. metabolism, inflammation, cytochrome p450 activity, oxidative stress or autophagy, etc.). Briefly, in these assays, reagents are added to the biochip that interact with cellular components of interest to luminesce, fluoresce or change the turbidity of the medium that is recorded by an in-system spectrophotometer.

[0246] FOURIER TRANSFORM INFRARED SPECTROSCOPY MEASUREMENT INTERFACE

[0247] In one embodiment of the operating system, ATR-FTIR is used as a measurement technique for cellular expression and functionality by delivering minute media samples from the biochip through a set of valves and tubing operated in a software-controlled sequence at multiple time points.

[0248] In another embodiment of the operating system, the ATR-FTIR performs tests to investigate the efficacy or toxicity of introduced drugs, including but not limited to element footprint, amide I, amide II, and molecular structure.

[0249] In another embodiment of the operating system, the ATR-FTIR is capable of detecting at least one sample having a thickness of at least 2.3 micrometers, the ATR-FTIR system includes at least one ATR crystal having a specific reflectance. The ATR crystal may have a reflectance in the range of 1.38 to 4.2. In an additional embodiment, the presently disclosed system can utilize sample aperture signal noise reduction or / and software based signal noise reduction by implementing a mapping mode with reference to a pre-created FTIR database.

[0250] In another embodiment of the operating system, the automatic ATR cover is equipped with a sensor to detect if the sample is properly placed in place. The ATR crystal cleaning system consists of valves, tubing and buffers controlled by the software. Once cleaning is complete, a test is performed by checking the reflectance and cross-referencing it with the actual refractive index. If the reflectance equals the actual reflectance, this means the ATR crystal is clean. If not, another cleaning cycle is performed by the operating system.

[0251] ATR-FTIR measures cellular response to drugs by providing quantitative data on drug uptake, metabolism and cellular expression. Selected data sets from the ATR-FTIR are stored in the operating system and then cross-referenced with other measurement techniques to further validate the results obtained.

[0252] It should be understood that various changes and modifications to the exemplary embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present subject matter and without diminishing its intended advantages. Accordingly, such changes and modifications are intended to be included within the scope of the appended claims. It should also be understood that the features of the dependent claims may be embodied in the systems, methods and apparatus of the respective independent claims.

[0253] Many modifications and other embodiments of the disclosures set forth herein will come to mind to one skilled in the art to which this disclosure pertains having the benefit of the teachings of the foregoing descriptions and the associated drawings. It is understood, therefore, that the disclosure is not limited to the particular embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

Claims

1. at least one human-on-a-chip comprising at least one organ-on-a-chip system including at least one ductal scaffold interacting with at least one surrounding compartment; at least one valve system controlling flow to each compartment; at least one micropumping mechanism capable of pumping and controlling flow to at least one compartment; at least one actuator that controls a valve on the chip and is coupled to the chip to control fluid transport to the chip compartment; Equipped with At least one valve system is operable to open, close and modify the flow paths to all compartments; Biochip fluid control system.

2. The at least one valve system is pneumatically, mechanically, electrically, and / or fluidically actuated. The biochip fluid control system of claim 1.

3. The at least one valve system connects at least one biochip to an operating system. The biochip fluid control system of claim 1.

4. The at least one valve system connects at least two biochips in a human-on-a-chip connection plate. The biochip fluid control system of claim 1.

5. The at least one valve system controls at least two biochips in a human-on-a-chip connection plate. The biochip fluid control system of claim 1.

6. At least one valve system controls at least one inlet and outlet of the biochip. The biochip fluid control system of claim 1.

7. The at least one valve system connects and controls the connection of at least one pipe to another pipe. The biochip fluid control system of claim 1.

8. The at least one valve system connects at least one interstitium to another interstitium. The biochip fluid control system of claim 1.

9. The at least one valve system connects and controls the connection of at least one tube to at least one interstitium. The biochip fluid control system of claim 1.

10. The at least one valve system connects the at least one biochip to the at least one measurement device. The biochip fluid control system of claim 1.

11. The at least one valve system includes at least one bistable valve and an on-off valve. The biochip fluid control system of claim 1.

12. The at least one valve system includes at least one bistable valve, the at least one bistable valve maintaining a state position by a beam, a magnet, or any drill geometry. The biochip fluid control system of claim 1.

13. The at least one valve system includes at least one on-off valve, and the at least one on-off valve maintains a state position according to a drill geometry. The biochip fluid control system of claim 1.

14. A human-on-chip plate for providing the at least one organ-on-chip system of the biochip fluid control system of claim 1, comprising: at least one 3D culture biochip; at least one microvalve; at least one microfluidic channel; at least one inlet or outlet plate port; Equipped with The outlet port from one chip is connected to another chip by routing valves; The chips are connected in parallel, series or in combination, and one or more chips in the human-on-chip plate are bypassed; Each fluid connection can be changed independently at any time. at least one valve system connects or separates different compartments of the organ-on-a-chip system; at least one inlet port and one outlet port configured to provide access to and isolation of the microchannel; the at least one valve system is capable of extracting, modifying a model flow map, and / or introducing or reducing at least one fluid chamber; The valve is controlled to a bistable position by an actuator and a sensor and is positioned at a channel port of the chip; The valve is placed directly next to the biochip and controlled to a bistable position. The human-on-a-chip plate further comprises at least one organ-on-a-chip system including at least one ductal scaffold interconnecting at least one surrounding compartment. Human-on-a-chip plate.

15. The plates are quickly connected from the bottom or the side, Fluids are exchanged, 15. The human-on-a-chip plate of claim 14.

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