Automatic bioreactor system, system for automatically implementing organ decellularization scheme, and waste purification system

Abstract

The application relates to an automated bioreactor system, an automated organ decellularization regimen system, and a waste purification system. An automated bioreactor system for decellularizing an organ includes a main chamber for containing the organ. The system also includes a reagent chamber containing a liquid phase reagent. A reagent conduit delivers a liquid phase reagent to the main chamber and a perfusion conduit delivers reagent from a reagent outlet in the main chamber into the organ. A perfusion pump drives a flow of reagent. A perfusion pressure sensor monitors the pressure of the flowing reagent. The control system controls the perfusion pump to drive the flow of reagent based on the received input representative of the desired pressure and the received input of the detected pressure. The control system may automatically perform all steps of the decellularization protocol based on sensor inputs. An automatic waste purification system may also be provided.

Description

[0001] This application is a divisional application of Chinese patent application No. 201580059153.6.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 044,647, filed September 2, 2014, which is incorporated herein by reference in its entirety. Field of the Invention

[0004] The systems and methods described herein relate generally to bioreactors and systems thereof, and more particularly, to automated bioreactor systems for organ decellularization, systems for automating organ decellularization protocols, and waste purification systems for bioreactors. Background Art

[0005] Although allogeneic lung transplantation is a treatment for advanced lung disease, the number of patients waiting for a lung transplant has steadily increased due to limited donor organs and only a small percentage of patients receive an organ transplant. For example, chronic obstructive pulmonary disease (COPD) affects more than 64 million people worldwide. The World Health Organization predicts that it will become the third leading cause of death by 2030. As another example, pulmonary arterial hypertension (PAH) affects the blood vessels in the lungs and can lead to right heart failure and death. Although there are FDA-approved treatments for PAH, there is still no cure, and for some patients, a lung transplant is the only option. Similarly, there are not enough organs to meet the demand.

[0006] Even in patients who receive an organ that can be used for transplantation, the clinical success rate of lung transplantation in patients is hampered by immunosuppression and chronic rejection, which can occur years after the patient has undergone an organ transplant. In the best case, patients suppress these problems by taking medications, which can come with their own serious side effects and last for the rest of the patient's life.

[0007] Tissue engineering provides an alternative to classical transplantation. This type of regeneration method has the potential to effectively bypass the limitations of tissue donor collections and prevent allogeneic rejection by providing a three-dimensional scaffold for inoculating autologous or stem cells that are specific to a particular patient. The technology includes treating the organ with a series of detergents, salts and / or enzymes to completely remove cellular material in the process of becoming decellularized, while leaving a complete extracellular matrix so that the matrix can be used as a scaffold for the subsequent regeneration of tissue. Decellularization generally includes continuous infusion of the organ with a series of detergents and repeated washings to remove residual DNA and other cell fragments from the organ. This can produce a scaffold in which extracellular matrix (ECM) proteins, organ structures and blood vessels are retained. In other words, the scaffold retains the structural features of the organ, but does not contain living cells or cell components. Due to the discovery that cell antigens that stimulate immune rejection are usually on the cell surface, removing such antigens can reduce the risk of rejection after the recellularized scaffold is implanted in the patient. For the lung, the decellularization process is particularly complex because it needs to retain the trachea and alveoli, as well as the pulmonary capillary bed to ensure the integrity of the gas exchange tissue.

[0008] Human intervention and manipulation during manually performed decellularization protocols increase the risk of contamination, reduce the consistency of the final product, and may adversely affect the three-dimensional structure and microarchitecture of the resulting scaffold, thereby wasting significant time and resources and resulting in a potentially useless product. SUMMARY OF THE INVENTION

[0010] There is a need to provide improved bioreactor systems and systems related thereto.

[0011] One embodiment of the present invention relates to an automated bioreactor system for decellularizing an organ, comprising a main chamber configured to contain the organ and having at least one reagent inlet, at least one reagent outlet, and at least one perfusion inlet. The system also includes at least one reagent chamber containing a liquid phase reagent and at least one reagent conduit configured to deliver the liquid phase reagent from the at least one reagent chamber to the main chamber through the at least one reagent inlet. At least one perfusion conduit is configured to deliver the liquid phase reagent to the organ from the at least one reagent outlet through the at least one perfusion inlet. At least one perfusion pump is configured to drive the liquid phase reagent to flow through the at least one perfusion conduit. At least one perfusion pressure sensor detects the pressure of the liquid phase reagent flowing through the at least one perfusion conduit. The control system receives an input representing the desired pressure of the liquid phase reagent flowing through the at least one perfusion conduit, receives an input of the pressure detected by the at least one perfusion pressure sensor, and outputs a signal to control at least one perfusion pump to drive the liquid phase reagent flow based on the received input representing the desired pressure and the received input of the detected pressure.

[0012] Another embodiment of the present invention relates to a system for automating a protocol for decellularizing an organ, comprising at least one perfusion pressure sensor configured to detect the pressure of a fluid flowing into the organ, at least one perfusion valve configured to control the fluid flowing into the organ, and at least one perfusion pump configured to drive the fluid flowing into the organ. The system also includes a control system configured to receive a protocol having a step of perfusing the organ, receive input from the at least one perfusion pressure sensor, and control the at least one perfusion valve and the at least one perfusion pump to automatically perform all steps of the protocol to decellularize the organ.

[0013] Another embodiment of the present invention is directed to a waste purification system for a biosensor, comprising a main chamber configured to contain an organ for perfusion, a waste chamber configured to receive waste fluid from the main chamber, and a waste conduit configured to deliver waste fluid from the main chamber to the waste chamber. The system also includes a purge fluid chamber configured to store a purge fluid and a purge conduit configured to direct the purge fluid from the purge fluid chamber to the waste fluid.

[0014] BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 is a schematic diagram of a bioreactor of an automated bioreactor system according to an exemplary embodiment.

[0016] Figure 2 is an exemplary embodiment of Figure 1 Block diagram of the bioreactor control system.

[0017] Figure 3 Yes Display Figure 2 A screen shot of one embodiment of a main screen of a control system's user interface. DETAILED DESCRIPTION OF THE INVENTION

[0019] Overview

[0020] The automatic bioreactor system described herein provides a bioreactor that can be set to automatically carry out all steps of the decellularization scheme, and the scheme after its startup is successfully executed with little or no user input. The automatic bioreactor system includes a main chamber where decellularization occurs and a series of reagent chambers that promote the reagent required for decellularization in a continuous closed conduit supplied to the main chamber. The control cell with a controller and a user section is incorporated into the automatic bioreactor system to fully automate the decellularization process in a closed system through valves and pumps, and the valves and pumps guide the reagent flow through the steps of the scheme. By providing full automation of the decellularization process, the direct interface between the components of the human and the bioreactor is reduced, thereby increasing the sterility, consistency and efficiency in the process, which improves the acquisition of more vigorous and structurally thorough decellularized organs for subsequent transplantation. In addition, the automatic bioreactor system allows a central control system that can simultaneously carry out multiple schemes between multiple bioreactors, further increasing the efficiency and consistency between multiple systems.

[0021] Although the process described below is in the context of a decellularization protocol for the lung, the automated bioreactor system described herein is not limited to such a protocol or organ. For example, the automated bioreactor system may also be used for other applications that may require perfusion of an organ. These applications may include, but are not limited to, processes in which decellularized lungs are required and any downstream analysis of the resulting scaffold material, recellularization of the scaffold, and engineering of a newly functional lung. In addition, the automated bioreactor system may also be used to perfuse any suitable organ or organ portion. Thus, the term "organ" includes an entire organ as well as any functional portion of an organ, such as a lobe.

[0022] Figure 1 An embodiment of an automated bioreactor system 100 for performing a complete decellularization protocol of porcine lungs is shown. Figure 1 The automated bioreactor system 100 generally comprises a main chamber 15 arranged to contain a lung 5 to be perfused, a chamber filling section 10 and a perfusion section 20. A waste purification system 30 may also be provided to the automated bioreactor system 100.

[0023] like Figure 2 As shown, the automated bioreactor system 100 also includes a controller 50, which is electrically connected to various components in various parts of the bioreactor and is configured to control them, as described in more detail below. In addition, a user interface 70 is additionally connected to and communicates with the controller 50, allowing the user to control the automated bioreactor system 100, as described in more detail below.

[0024] main chamber

[0025] The main chamber 15 can be any suitable chamber that can produce and maintain the aseptic sealed environment for perfusing organs or lungs 5. The main chamber 15 preferably comprises a reagent inlet 14, a reagent outlet 23, a perfusion inlet 24 and a waste outlet 31, 32, which are described in more detail below. In addition, the main chamber 15 is located at the top of a weight sensor 60, which is described in more detail below. In some embodiments, at least part of the cavity of the main chamber 15 is transparent or translucent so that the user can observe the organ during the use of the bioreactor system. In some embodiments, the main chamber 15 will have an entrance for operating the main chamber interior during use. This operation can be carried out by physically inserting objects such as tools and equipment into the main chamber 15, or the interface of the object already present in the user's operation chamber can be provided. For example, an entrance can be designed so that the main chamber 15 can be used as a glove box.

[0026] Chamber filling part

[0027] like Figure 1 As shown, the chamber filling portion 10 preferably includes a plurality of reagent chambers 11, each of which is fluidically connected to the main chamber 15 by a reagent conduit, which fluid conduit may be in the form of a pipe 40. Although a plurality of reagent chambers 11 are shown, a smaller or greater number of chambers may be used. The reagent chamber 11 is configured to hold a specific type of liquid reagent required for a given step in the decellularization protocol. The liquid reagent may be in the form of a solution, an emulsion, or a suspension. The reagent chamber 11 may be any suitable container for holding reagents, such as a large glass, a syringe, and the like. In addition, the reagents contained in each of the reagent chambers 11 may be any suitable reagent required for organ decellularization, such as phosphate buffered saline (PBS), sodium dodecyl sulfate (SDS), Triton X-100, and the like. The conduit 40 of the reagent conduit may include any conventional material suitable for transferring reagents and biological materials and for aseptic connections between unknown chambers, such as silicon tubing, and the like. As Figure 1 As shown, a reagent conduit, such as tubing 40 , allows liquid reagent to flow into the main chamber 15 through connection with the reagent inlet 14 .

[0028] A reagent valve manifold 12 and a reagent pump 13 are placed between the reagent chamber 11 and the main chamber 15. The reagent valve manifold 12 controls the flow of liquid phase reagents from a given reagent chamber 11 to the main chamber 15 containing the lung 5. The reagent valve manifold 12 includes a plurality of valve ports. The number of valve ports provided on the reagent valve manifold 12 can be any suitable number to allow the numerous liquid phase reagents required by the protocol to flow in-line. Figure 1As shown, the reagent valve manifold 12 includes 5 valve ports so that up to 5 reagent chambers 11 can be controlled to flow through the pipeline 40 connected to the reagent inlet 14 of the main chamber 15. The reagent valve manifold 13 can also be any suitable valve mechanism, which can control the liquid phase reagent to flow from a specific reagent chamber 11 to the chamber filling inlet 14 of the main chamber 15. Preferably, the reagent valve manifold 13 includes a reagent valve in the form of a solenoid clamp type valve mechanism. These solenoid clamp type valve mechanisms can control the liquid phase reagent to flow through the pipeline 40 by applying force to the pipeline 40 to stop the liquid phase reagent from flowing, and they are flexible when using the solenoid clamp type valve mechanism. This valve mechanism is an external cut-off device, which minimizes the contact with the reagent contained in the pipeline 40, thereby increasing the sterility of the system.

[0029] The reagent pump 13 is connected to the conduit 40 downstream of the reagent valve manifold 12. The reagent pump 13 drives the liquid phase reagent from a given reagent chamber 11 into the reagent inlet 14. The reagent pump 13 can be any suitable mechanism for pumping fluid into the main chamber 15. Figure 1 In the illustrated embodiment, the reagent pump 13 is a double-head peristaltic pump to allow for increased flow rates and pressures and to maintain the sterility of the fluid in the tubing 40. Figure 2 As shown, the reagent valve manifold 12 and the reagent pump 13 are both electrically connected to the controller 50 and are configured to be controlled by the controller 50 .

[0030] As mentioned above, the main chamber 15 is located on a weight sensor 60, such as a scale. The weight sensor 60 is configured to detect the weight present in the main chamber 15. Figure 2 As shown, the weight sensor 60 transmits the detected weight to the controller 50 .

[0031] Perfusion system

[0032] An embodiment of the perfusion system 20 is shown in Figure 1 It is preferably arranged as a closed circulation system with a perfusion conduit, preferably in the form of an additional conduit 40, leading from the reagent outlet 23 of the main chamber 15 to the perfusion inlet 24 of the main chamber 15, so that the liquid phase reagent contained in the main chamber 15 can be recirculated and perfused through the lung 5. The conduit 40 forming the perfusion conduit can also extend through the perfusion inlet 24 to the main chamber 15 and to the lung 5 contained in the main chamber 15. Figure 1 As shown, tubing 40 may be configured to extend to the pulmonary artery of lung 5 so that perfusion may occur through the cardiopulmonary circulation of lung 5 .

[0033] The liquid reagent is driven to flow through the lung 5 by the perfusion pump 21. Figure 2 As shown, it is electrically connected to the controller 50 and is configured to be controlled by the controller 50. Figure 1As shown, the perfusion pump 21 can be a single-head peristaltic pump. A pulse buffer 22 can be included and located between the perfusion pump 21 and the main chamber 15. The pulse buffer 22 can be used to prevent bubbles from entering the liquid phase reagent before perfusion to the lung 5, or can be used to prevent damage to the lung 5 by flow pulses that may be generated by the perfusion pump 21 during operation. The pulse buffer 22 can be any suitable closed container configured to contain residual fluid circulation in the perfusion system. In addition, a stirring rod can be provided to the pulse buffer 22 to reduce the probability of stagnation in the holes of the pulse buffer 22, which may cause contamination. In addition, the pulse buffer 22 can reduce the noise in the pressure output waveform in the liquid phase reagent flowing to the lung 5. This in turn can make the pressure more accurately measured by the perfusion pressure sensor 25, which is described in more detail below.

[0034] Connected to the tubing 40 upstream of the perfusion inlet 24 of the main chamber 15 is a perfusion pressure sensor 25 for detecting the pressure of the liquid phase agent flowing through the tubing 40 and into the lung 5. Figure 2 As shown, the perfusion pressure sensor 25 is also configured to communicate the detected pressure of the pressurized reagent to the controller 50. The perfusion pressure sensor 25 can be configured to detect the pressure of the liquid phase reagent at any point downstream of the perfusion inlet 24. Figure 1 As shown, the perfusion pressure sensor 25 is connected to the tubing 40 so that the perfusion pressure sensor 25 detects the pressure at the perfusion inlet 24. The perfusion pressure sensor 25 may be placed outside the chamber at a height that approximately matches the height at which the lung 5 floats in the main chamber 5 during perfusion.

[0035] Waste purification system

[0036] like Figure 1 As shown, a preferred embodiment of the waste purification system 30 generally includes a purification fluid chamber 34 and a waste chamber 35. The waste chamber 35 is fluidly connected to receive waste fluid from the main chamber 15 through a waste conduit, and the purification fluid chamber 34 is fluidly connected to the waste chamber 35 through a purification conduit, which can be formed by an additional pipe 40. The purification fluid chamber 34 is configured to hold a purification fluid, such as bleach, and the waste chamber 35 is configured to hold a mixture containing waste fluid from the main chamber 15 and purification fluid from the purification fluid chamber 34.

[0037] like Figure 1 As shown, two drain valves 33 are placed between the main chamber 15 and the waste chamber 35, which control the flow of waste fluid from the main chamber 15 through the waste outlets 31, 32. As described above for the valve manifold 12, the drain valves 33 may also include a spiral clamp valve as a valve closure mechanism for enhanced sterility of the closed system. In addition, each waste outlet 31, 32 may also be provided with a one-way valve to prevent the waste fluid from flowing back into the main chamber 15.

[0038] Connected to the downstream of the drain valve 33 is a purge pump 36. Figure 2 As shown, the purge pump 36 is electrically connected to the controller 50 and is controlled by the controller 50. The purge pump 36 drives the waste fluid from the main chamber 10 to the waste chamber 27. In addition, the purge pump 36, such as Figure 1 As shown, the purified fluid can also be driven to flow from the purified fluid chamber 34 to the waste chamber 35. Figure 1 As shown, the purification pump 55 can take the form of a 4-head peristaltic pump, wherein two pump heads are configured to drive the waste fluid flow in the pipe 40 connecting the main chamber 15 to the waste chamber 35, and the other two pump heads are configured to drive the purification fluid to flow from the purification fluid chamber 34 to the waste chamber 35.

[0039] In some cases, certain purification schemes require that the waste fluid be mixed with a specific proportion of the purified fluid before the waste fluid can be safely and properly discharged. For example, the waste fluid may be mixed with a desired proportion (e.g., 10%) of the total volume of the waste fluid bleach prior to discharge. To this end, in one embodiment, the diameter of the purification conduit leading from the purified fluid chamber 34 to the waste chamber 35 is smaller than the diameter of the waste conduit leading from the main chamber 15 to the waste chamber 35 by a desired proportion (e.g., 1 / 10 the size). And, as Figure 1 As shown, since the purge pump 36 drives the waste fluid flow from the main chamber 15 and the purge fluid flow from the purge fluid chamber 34, the purge pump 36 can be set to pump at a constant flow rate so that the purge fluids can be mixed in the appropriate ratio of the fluid rate flowing into the waste chamber 35. In addition, as shown in FIG. Figure 1 As shown, to ensure adequate mixing of the purge fluid with the waste fluid, the purge fluid may be introduced into the waste fluid in-line within the waste conduit and upstream of the waste chamber 35. However, the purge fluid may also be introduced directly into the waste fluid and subsequently mixed within the waste chamber 35.

[0040] Additionally, separate pumps may be used to drive waste fluid from the main chamber 15 to the waste chamber 35 and purge fluid from the chamber 34 to the waste chamber 35. In this sense, the controller 40 may then control each purge pump 36 and other pumps to drive waste fluid from the main chamber 10 and purge fluid from the purge fluid chamber 34, respectively. Each pump may be controlled to operate at a corresponding flow rate so that the purge fluid is mixed with the waste fluid in a ratio required for safe and proper discharge.

[0041] like Figure 1 As shown, a drain conduit in the form of an additional pipe 40 extends from the waste chamber 35 to the drain pipe 37. Located between the waste chamber 35 and the drain pipe 37 is a drain pump 38, which drives the waste fluid and purified fluid mixture from the waste chamber 35 to the drain pipe 37. Figure 2As shown, a drain pump 38, which may be a double-headed peristaltic pump, is electrically connected to and controlled by the controller 50. Alternatively, however, the drain pump 38 may be omitted and a drain conduit may instead extend from the bottom of the waste chamber 35. A valve, such as a solenoid pinch valve, may be included downstream of the outlet of the waste chamber 35. The controller 50 may then be arranged to control the valve so that, when open, the waste fluid and purge fluid mixture flows to the drain 37 by gravity.

[0042] like Figure 1 Additionally, a plurality of secondary containment chambers 39 are shown, which serve as a secondary measure to prevent waste fluid from flowing out of the automated bioreactor system 100. Figure 1 As shown, the main chamber 15, the purified fluid chamber 34, and the waste chamber 35 are each placed in a secondary containment chamber 39. The secondary containment chamber 39 is used to contain any overflow of fluid from each of these chambers. Figure 1 As shown, each secondary containment chamber 39 is connected to a plurality of drain conduits, each in the form of a pipe 40 that allows any fluid contained in the secondary purification chamber 39 to flow into the drain 37 .

[0043] The automated bioreactor system 100 is not limited to Figure 1 The sensors shown in the figure may include any other sensors that can measure variables related to the perfusion process. For example, a temperature control system may be included. The temperature control system may have a temperature sensor placed adjacent to the main chamber 15 to detect the temperature of the liquid phase reagent and the lung 5 contained in the main chamber 15. The temperature control system may also include a heating and / or cooling element, which is configured to heat and / or cool the temperature of the liquid phase reagent and the lung 5 contained in the main chamber 15. The temperature sensor may then be configured to interact with the controller 50 to detect the temperature. The controller 50 may then be configured to control the heating and / or cooling element to adjust the temperature of the liquid phase reagent and the lung 5 to a safe temperature for the decellularization process. By controlling the temperature in the main chamber 15, the sterility of the system and the structural integrity of the organ can also be further maintained. Other sensors may also include, but are not limited to, pH sensors, conductivity sensors, etc.

[0044] As described above, the automated bioreactor system 100 includes a bioreactor for decellularizing lungs in a closed sterile environment. By limiting human interaction with the mechanisms controlling the decellularization process and by minimizing contact between the moving parts of the system (e.g., valve mechanisms, pumps) and the internal components (e.g., liquid reagents, organs), a more sterile bioreactor system can be achieved, thereby increasing the likelihood of a viable and well-structured scaffold after the decellularization process.

[0045] Control Process

[0046] Now the continuous reference Figure 1 and2 An overview of the control process for automating the entire protocol for decellularizing lung 5 is described. Generally, the control system can automatically implement all steps of the entire protocol by performing three types of processes sequentially and / or simultaneously: chamber filling process, perfusion process, and waste purge process.

[0047] The protocol can be any known protocol for perfusing or decellularizing an organ, as well as a new protocol. The protocol can be provided to the system in any of a variety of ways. For example, the protocol can be uploaded as part of a data spreadsheet. Alternatively, the protocol, or multiple protocols, can be stored on the system for selection by the user. As another alternative, the stored protocol can be modified by the user.

[0048] The control system, or its controller 50, can be a general purpose processor, a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA) or other programmable logic device, discrete gate or transistor logic elements, discrete hardware components, or any combination thereof designed to perform the functions described herein. The general purpose processor can be a microprocessor, but alternatively, the processor can be any conventional processor, controller, microcontroller, or state machine. The processor can also be implemented as a combination of computing devices, for example, a DSP and a microprocessor, a plurality of microprocessors, one or more microprocessors and a DSP core, or any other such configuration. The steps of the algorithm or method described in association with the embodiments described herein can be directly embodied in hardware, in a software module executed by a processor, or a combination of both. The software module can be in RAM memory, flash memory, ROM memory, EPROM memory, EEPROM memory, cache, hard disk, removable disk, CD-ROM, or any other type of storage medium known in the art. An exemplary storage medium is coupled to the processor so that the processor can read information from the storage medium and write information to the storage medium. Alternatively, the storage medium can be integrated into the processor. The processor and the storage medium can be in an ASIC. The ASIC can be in a user terminal. Alternatively, the processor and storage medium can be located at the user terminal as discrete components. In one or more exemplary embodiments, the functions described can be implemented in hardware, software, firmware, or any combination thereof. Such hardware, software, firmware, or any combination thereof can be part of or implemented by any one or combination of a processor, a database, a related component in a system, its components, etc., or a processor, a database, a related component in a system, its components, etc. If implemented in software, the function can be stored or transmitted by one or more instructions or codes on a computer-readable medium. Computer-readable media include computer storage media and communication media, including any media that facilitates the transfer of computer programs from one place to another. Storage media can be any accessible medium that can be accessed by a computer. For example, but not limited to, such a computer-readable medium may include RAM, ROM, EEPROM, CD-ROM or other optical disk storage, disk storage or other magnetic storage device, or any other medium that can be used to carry or store the required program code in the form of an instruction or data structure and can be accessed by a computer. In addition, any connection is appropriately called a computer-readable medium. For example, if software is transmitted from a web page, server, or other remote source using coaxial cable, fiber optic cable, twisted pair, digital subscriber line (DSL), or wireless technologies such as infrared, radiation, and microwave, then the coaxial cable, fiber optic cable, twisted pair, DSL, or wireless technologies such as infrared, radiation, and microwave are included in the definition of medium.Disk and disc as used herein include compact disc (CD), laser disc, optical disc, digital versatile disc (DVD), floppy disk, and Blu-ray disc, where disks usually reproduce data magnetically, while discs reproduce data optically with lasers. Combinations of the above should also be included within the scope of computer-readable media.

[0049] The chamber filling process starts from the controller 50 outputting a signal to the reagent valve manifold 12 to open one or more of the valves (at the same time, closing any valve not required by the process) to allow the liquid phase reagent contained in a given reagent chamber 11 to flow. At the same time, the controller 50 also outputs a signal to the reagent pump 13 to drive the liquid phase reagent to flow from the reagent chamber 11 connected to the valve line now opened into the pipeline 40 leading to the reagent inlet 14. The reagent pump 13 can be driven according to a specific scheme and / or user settings. For example, the controller 50 can output to the reagent pump 13 to pump at a constant flow rate (e.g., 100 ml / min) or a constant pressure (e.g., 30 mmHg). If the reagent pump 13 is a peristaltic pump, the controller 50 can also output to the reagent pump 13 to run in a given direction (e.g., clockwise or counterclockwise).

[0050] When the main chamber 15 is filled with the required liquid phase reagent, the controller 50 continuously receives input of the weight of the main chamber 15 detected by the weight sensor 60, such as Figure 2 As shown. Using this detected weight, the controller 50 then determines the volume of the liquid phase reagent present in the main chamber 15 at that given time. For example, the controller 50 can determine the volume of the liquid phase reagent based on the detected weight of the main chamber 15 and the known density of the liquid phase reagent used, which can be pre-stored in the program before implementing the protocol. By continuously detecting the weight of the main chamber 15 and the volume of the liquid phase reagent present in the main chamber 15, the controller 50 can then determine when the volume of the liquid phase reagent required for the chamber filling step of the protocol has been reached.

[0051] In addition, the controller 50 may also be configured to determine whether and / or when the maximum volume of a liquid reagent has been introduced into the main chamber 15. In this case, before implementing the scheme, the controller 50 may be programmed with a pre-stored threshold volume, which may represent the maximum volume of the fluid that the main chamber can hold, or some proportions of this maximum volume. During the implementation of the chamber filling step, if the controller 50 determines that the main chamber 50 has been filled, thereby reaching the threshold volume, the processor 50 may be configured to stop the chamber filling step to continue (by stopping the operation of the reagent pump 13) even if the chamber filling step still needs to introduce additional liquid reagent into the main chamber 15. The controller 50 may then automatically proceed to the next step (e.g., a priming step or a draining step) in the scheme. In this sense, by continuously monitoring the weight of the main chamber 10, the controller 50 may place the overfilling of the main chamber 15, thereby providing protection for the system.

[0052] During the perfusion process, all valves in the system are closed, and the controller 50 outputs a signal to control the perfusion pump 21 to drive the liquid phase reagent contained in the main chamber 15 to flow into the lung 5 (e.g., into the pulmonary artery of the lung 5). Similar to the above-mentioned chamber filling process, the controller 50 can output a signal to control the perfusion pump 21 to drive the liquid phase reagent to flow at a constant pressure or a constant flow rate, as defined by the scheme. In addition, if a peristaltic pump is used, the controller 50 can guide the perfusion pump 21 to also run in a given direction.

[0053] During perfusion, the controller 50 continuously receives input of the pressure of the liquid agent flowing to the lung 5 detected by the perfusion pressure sensor 25, such as Figure 2 As shown. The controller 50 may also be configured to receive a liquid phase agent input into the lung 5 at a desired pressure and / or flow rate. The desired pressure and / or flow rate may be defined by a protocol or may be specifically specified by a user and may be input into the control system by an input representing the desired pressure through the perfusion catheter. Based on the received detected pressure and the received input representing the desired pressure, the controller 50 may then output a signal to control the perfusion pump 21 to continuously maintain the desired pressure and / or flow rate entering the lung 5. The controller 50 may also be configured to detect whether and / or when the detected pressure deviates from the desired pressure and / or flow rate of the liquid phase agent entering the lung 5 by a pre-stored threshold amount. For example, the controller 50 may be configured to detect whether and / or when the detected pressure is 15% or 15% higher or lower than the pressure and / or flow rate defined by the protocol. The controller 50 may then be further configured to communicate this deviation with the user interface 70, which may then warn the user of the deviation for troubleshooting purposes.

[0054] After completing the priming step, the controller 50 may be configured to initiate an additional wash step using, for example, water or saline to wash away residual liquid reagent from the lung 5 or the perfusion catheter before proceeding to the next step. In this case, the controller 50 may output a signal to the valve manifold 14 of the chamber filling portion 10 to open the valve and enable flow from the reagent chamber 11 configured to hold the washing fluid. The controller 50 may then control the reagent pump 13 and the perfusion pump 21 to initiate the flow of the washing fluid through the system.

[0055] During the waste purge process, the controller 50 outputs a signal to open the drain valve 33 to allow the waste fluid to flow from the main chamber 15 to the waste chamber 35. At the same time, the controller 50 outputs a signal to the purge pump 36 to direct the waste fluid to flow from the main chamber 15 to the waste chamber 35. Figure 1As shown, the purge pump 36 may also be configured to direct the purge fluid from the purge fluid chamber 34 to the waste chamber 35. The controller 50 may then output a signal to the purge pump 36 to direct the waste fluid and the purge fluid to flow through the conduit 40 so that the two fluids are mixed in a suitable ratio. Alternatively, if two separate pumps are used to direct the waste fluid flow and the purge fluid flow, respectively, the controller 50 may output a signal to each pump to direct the corresponding fluid to flow at a rate that will ensure proper mixing of the two fluids. As similarly described above, the controller 50 may output a signal to the purge pump 36 to pump the waste fluid and / or the purge fluid through the conduit 40 at a constant pressure or a constant flow rate.

[0056] As the waste purification process proceeds, the controller 50 continuously monitors the volume of waste fluid removed from the main chamber 15 by receiving a weight output detected by the weight sensor 60. Once the desired volume of waste fluid has been removed from the main chamber 15, as defined by the protocol, the controller 50 outputs a signal to the valve 33 to close. Once the waste fluid and purified fluid mixture has been mixed and settled in the waste chamber 35 for the desired time to ensure adequate purification, the controller 50 then outputs a signal to the drain pump 38 to drive the fluid contained in the waste chamber 35 into the drain pipe 37 for drainage. Alternatively, as described above, the controller 50 may be configured to output a signal to open a valve located at the bottom of the waste chamber so that the fluid contained in the waste chamber 35 is drained by gravity.

[0057] As mentioned above and as Figure 2 As shown, the controller 50 communicates with a user interface 70. The user interface 70 enables a user to remotely program a decellularization protocol for upload to the control system, and then set up and control an automated protocol executed by the automated bioreactor system 100. An embodiment of the main screen of the user interface 70 is shown in Figure 3 and is described in further detail below.

[0058] The user interface 70 enables the user to define a series of steps (a "recipe") containing operational information for the pump and valve functions. The user defined recipe is used to direct the controller 50 to the appropriate components to control at a given time during the execution of the overall protocol. This series of steps may be written by the user using any suitable text-based program (either remotely or within the user interface 70), which may then be uploaded to the control system for execution. In addition, the defined series of steps may be stored in the control system for later recall and execution, thereby making it easy to select and implement commonly used protocols and settings.

[0059] Prior to commencing the automated decellularization process, the user interface 70 enables the user to set the system settings for a given bioreactor for a decellularization protocol under system setup instructions 89, which may bring up a separate window of the user interface 70 to allow the user to establish the settings for the bioreactor system. Here, pressure inputs, digital outputs of the solenoid mounts, and pump assignments may be set here to ensure that data is read appropriately from the device. Once the desired system settings have been selected by the user, the user may then drive the automated bioreactor system 100 to execute the selected protocol via a start instruction through the user interface 70. A restart hardware instruction 90 may also be included to prime the hardware prior to commencing a new recipe.

[0060] During the execution of the automated decellularization process, the user interface 70 enables the user to remotely monitor and control the execution of the protocol. Figure 3 In section A of the main screen of the user interface 70, the user can start, stop or pause execution of the protocol while it is in progress using command button 71. Although pausing execution using command button 71 temporarily stops all pumps, the user can still turn on the pumps using the override parameters contained in section E, which are described in further detail below. In addition, command button 71 can also include a skip command that allows the user to skip the current step in the protocol and proceed to the subsequent step.

[0061] The user interface 70 may also provide the user with real-time information about the status of the bioreactor as the protocol is executed. Figure 3 As shown, under section C, under indication 73, the user interface 70 can allow the user to see the remaining time to execute the current step and / or the overall protocol. The user interface 70 can also provide a visual list of all the steps of the protocol under label 72 and display the current step being executed by the system under indication 74. In addition, the user interface 70 can allow the user to monitor the input received by the controller 50 from various sensors present in the system. For example, the user interface 70 can be configured to display a waveform table 75, under which the user interface 70 can be configured to display a waveform table 75. Figure 3Under the portion D of the main screen shown, it displays the pressure detected by the pressure sensor 25 over time. Aspects of the waveform table can be operated by the user with suitable drawing operation tools (e.g., zoom, ruler, drag, etc.). The measured values ​​of the pressure and flow rate determined by the controller 50 based on the detected pressure input received from the pressure sensor 25 are also displayed by the table 76. In addition, for systems using multiple pressure sensors, a toggle button 87 can be provided so that the user can switch and / or add real-time information about the pressure detected by each pressure sensor. Similarly, the volume of the liquid phase reagent contained in the main chamber 15 determined by the controller 50 can also be displayed in real time by the indication 77 under the portion D. The user can use the recalibration instruction 88 to zero and / or calibrate any pressure sensor. However, preferably, the recalibration instruction 88 is only active before the recipe starts and becomes inactive when the automatic cell removal scheme is active.

[0062] In addition, the user interface 70 may also be configured to inform the user of the current operating status of the valves and pumps. For example, the user interface 70 may display to the user the current operating status of a given pump (e.g., idle, test, alarm status) under a label 78. Under this label, errors for solenoids, pumps, and pressure inputs are also displayed. Scale debug values ​​may also be displayed, including measurements for displaying volumes and for calculating the volume of a fill step.

[0063] The overall main status of the system (e.g., idle, test / run, alarm status) may be indicated to the user via a label 79 under section C of the main screen of the user interface 70. As described above, the controller 50 may be configured to detect whether the detected pressure of the flow rate driven by a given pump deviates beyond a predetermined threshold. If such a deviation occurs, the controller 50 may then communicate this status to the user interface 70, which may be configured to alert the user to the alarm status of the pump (e.g., the indicator 79 changes from green to yellow when the pressure reading deviates from the pressure set point of at least one pump by 15%, and then changes from yellow to red when the pressure reading deviates from the pressure set point of at least one pump by more than 30%).

[0064] For troubleshooting purposes, the user interface 70 may include Figure 3 The override control parameters are under section E of the main screen shown. In this section, the user can change the pump settings and value states during the execution of the regimen to deviate from the uploaded regimen. For example, the pump settings can be overridden through the pump control section 82. Using the pump control section 82, the user can turn the pump on or off, change the pump's operating mode (e.g., constant flow rate to constant pressure), change the pump's operating direction (e.g., clockwise to counterclockwise), and / or change the flow rate and / or pressure set points. Similarly, valve settings can also be changed under the valve control section 83, where the user can open or close specific valves in the system. Fluid settings can also be manually changed through the fluid control section 84. As shown Figure 3As shown, the user can select a fluid from a library of drop-down menus that corresponds to the fluid flow controlled by the valve shown in the valve control portion 83. Alternatively, the user can choose to manually insert a specific fluid that is not included in the drop-down menu, and the system can be updated accordingly using the fluid setup instructions 85. Once the selection is updated, the density information can then be passed to the control system for calculating the displayed volume and target fill volume of the main chamber 15. Container setup instructions can also be included to allow the user to enter information about the bioreactor itself. Thus, the user interface 70 enables continuous monitoring and control of the automated decellularization process to enable more effective and efficient control of the overall protocol.

[0065] The user interface 70 may also include notification settings instructions that enable a user to remotely receive notifications about the system as the system executes the protocol. The settings instructions 81 may provide automated notifications by the control system, such as in the form of emails, of any changes in a given step and periodic updates on the system status as a whole. In addition, the user interface 70 may insert time-stamped comments into the data directory via annotation instructions 86, which are saved in the system for later viewing. This allows the user to note specific changes made to the protocol for analysis purposes, such as inserting instructions about steps taken during setup, which may correspond to anomalies in the data directory, for example, by adjusting a pressure sensor.

[0066] The user interface 70 can also monitor and control the execution of multiple protocols each executed in a separate bioreactor, such as Figure 1 As shown. In this case, the controller 50 can be used as a central control system and control the components of each bioreactor according to each series of steps determined by the user. The user interface 70 allows the user to switch between the executed protocols to monitor and control each separate protocol simultaneously. Through a centralized system, several decellularization protocols can be performed simultaneously, thereby increasing the efficiency of the entire process.

[0067] Exemplary Protocol for Preparation of Lung Tissue to be Decellularized by a Bioreactor System

[0068] Prepare a clean work area in a biological safety cabinet (BSC). Perform all steps of the preparation process using good aseptic technique to minimize the introduction of environmental microbial contaminants. Use sterile sleeves and gloves and exchange gloves as needed.

[0069] The internal organs (i.e., cardiopulmonary circulation) are removed from the shipping container and the trauma of blood vessels, trachea, pleura and substance are strictly checked. The internal organs can then be properly dissected under aseptic conditions. The packaging can be sterilized. The pulmonary artery catheter is inserted into the pulmonary artery and fixed, and the internal organs are dissected to remove the myocardium. Any remaining heart tissue can be trimmed. The lungs can then be perfused with heparinized saline through the pulmonary vein for anticoagulation during the subsequent perfusion in the bioreactor system. The prepared lungs are then weighed and transferred to the fume hood area for loading into the main chamber when needed.

[0070] Exemplary setup of a bioreactor in a fume hood

[0071] With the fume hood blower turned on, spray the fume hood down with an appropriate cleaning solution. Place the autoclaved main chamber near the fume hood with the outer layer of the autoclave wrap unwound. Use the ramp to slide the main chamber into the fume hood. Using a sleeve and surgical gloves, unwrap the inner layer of the autoclave wrap on the main chamber and remove from the fume hood. Remove the spacers between the bioreactor glass and the base. Once all spacers are removed, verify that the glass is resting on the bottom gasket with an even edge around the bottom.

[0072] The autoclave wrap covering the glass is removed. A person outside the fume hood removes the outer layer of the top ring and brings it into the fume hood using sterile gloves. The sterile drape is brought into the fume hood. The top ring is then secured to the metal copper post using screws and tools.

[0073] Bring the main chamber lid into the fume hood. Place the lid on top of the main chamber and attach the lid to the top ring with fasteners. The main chamber is now closed to the environment and all ports on the base and lid are capped. Using a sterile biowelder, connect the reagent lines to the reagent tubing assembly. This assembly connects to the reagent chamber. Use a peristaltic pump to fill the main chamber.

[0074] After filling to the tube drain level, stop the reagent pump. The perfusion tubing is connected from the base to one end of the pulse buffer. The other side of the pulse buffer is connected to the perfusion tubing at the top of the bioreactor to complete the perfusion circuit.

[0075] Initiate perfusion by infusing the solution through the perfusion catheter and remove air from the line using a sterile syringe. Fill the main chamber to approximately 20 L.

[0076] Exemplary protocol for aseptic mounting of a lung in a fume hood

[0077] Run the perfusion pump slowly (25 mL / min) and purge all air from the perfusion circuit. Bring the organ (lung) in the covered sterile tray into the fume hood. Release the crank on the main chamber lid. Using sterile gloves, move the lung into the main chamber and make the connection.

[0078] Assess any air bubbles in the perfusion catheter.Use the reagent pump to pump the solution back into the 1x PBS reagent chamber until the main chamber is about 20L.The reagent catheter is sealed and disconnected from the 1x PBS reagent chamber.Then remove the main chamber in the fume hood and transfer to the decellularization station.Disinfect the fume hood with a suitable disinfectant after installing the lung.

[0079] Exemplary Automated Lung Decellularization Protocol

[0080] Once the main chamber is on the balance, use a sterile pipette to connect the following lines and complete the bioreactor fluid circuit: (1) reagent line; (2) drain line; (3) reagent chamber connection to the valve manifold.

[0081] Then start perfusion manually at a low rate. Manually fill the main chamber with approximately 1-2 L of PBS at a rate of 1000 mL / min.

[0082] Place the perfusion pressure transducer outside the chamber at a height of 40 L, matching the height at which the lungs will float during most of the experiment. Zero the perfusion pressure by pressing the "Recalibrate" button, then zero pressure transducer 1 (whichever transducer is controlling the perfusion pump, as specified by the recipe).

[0083] The experiment was started by pressing the "Start" button, which prompted for the appropriate recipe file, data storage unknown, and solution / chamber settings.

[0084] The automated decellularization process for lung accessible samples was as follows: (1) perfusion with PBS at 30 mmHg for 2 hours, draining 20 L from the main chamber once formulation began; (2) 5 cycles of chamber filling, perfusion, and draining with 0.5% SDS over 5 days; (3) one cycle of chamber filling and draining, and one day of exposure to DiH2O; (4) 1 cycle of chamber filling, perfusion, and draining with 0.5% Triton-X over 1 day; and (5) a final wash step of 5 cycles of chamber filling, perfusion, and draining with PBS over 5 days.

[0085] Frequent monitoring of tissue status and bioreactor function is important for the process; depending on tissue perfusion efficiency, manual intervention for overflow waste removal may be required. Sampling of chamber fluids can be performed during sterility testing or analytical procedures. These samples are obtained through swabbable ports on the top of the main chamber.

[0086] Exemplary Storage of Decellularized Tissue

[0087] At the end of the experiment, the decellularized tissue will be stored, sampled, and formaldehyde fixed or both.

[0088] The experiment is stopped on the software interface by pressing the "Stop" button. The main chamber is manually drained (waste first, if necessary) until there is no liquid remaining in the main chamber. All connections from the main chamber to the peripheral tubing are closed using the Clipster tool or the heat sealing tool.

[0089] Sterilize the exterior of the main chamber before placing it in the fume hood. Load the main chamber into the fume hood and then sterilize the tray / drape. Remove the lid. Using sterile gloves, disconnect the lungs from the perfusion tubing and bring to the tray. Remove the tray from the fume hood and bring into the biosafety cabinet for storage or sampling. Remove the main chamber from the fume hood and sterilize the unit according to the protocol.

[0090] Exemplary Protocols for Chamber and Station Cleaning

[0091] All tubing and fittings downstream of the chamber (drain tubing, waste) should be disposed of in a biohazardous waste container. The irrigation catheter tubing should also be disposed of in this manner; however, the pressure sensor may be cleaned and retained.

[0092] Due to the relative fragility of the coating on the main chamber and its components, the main chamber components must be manually washed with a mild detergent. The clean main chamber components are dried.

[0093] The waste chamber is stripped of its external tubing - this tubing is disposed of as biohazardous waste. The carboy itself can be washed in a parts washer. The cap / seal tube in the waste drain carboy should be washed manually. The overflow containment container that holds the waste drain carboy can be washed manually if there is a spill during the decellularization experiment. The bleach carboy does not need to be washed, only refilled with bleach when appropriate. The bleach line loop should be replaced for each decellularization to avoid cracking of fittings that come in contact with the bleach.

[0094] The pulse dampener should be cleaned of residual fluid with bleach and poured down a drain. The pulse dampener itself can be disassembled and washed. The stirring rod can be washed manually or in a basket in a parts washer.

[0095] Exemplary Operation of Bioreactor Procedure

[0096] Open the Bioreactor program from the computer desktop and update the system settings if hardware or software changes have occurred. Under the system settings, the following four properties must be set for normal decellularization to ensure that data will be read appropriately from the device: (1) Analog Input Device (Pressure Input); (2) DigOut Device (Solenoid); (3) Analog Output Task (Pump Control); and (4) Balance COM Port (Balance-Port Connection). Flow rate limits are also set to determine the minimum and maximum flow rates in the perfusion line when the perfusion pump is pressure controlled. Use the PC ID as an identifier to retain system parameters. For example, when the PCID is set to "Bioreactor 1", all related settings are retained to that ID. The last PC ID (and related system settings) used for the program execution will default to subsequent executions of the program. PID parameters default to Kp, Ki, and Kd values ​​of 0.1. The median filter rank is a filter that averages the data point readings from the pressure sensor to minimize noise. The larger the specific value, the greater the filter effect observed. It defaults to a value of 2, but can be modified to reduce noise in the pressure gauge. A value of 25 has been found to be suitable for automated decellularization processes.

[0097] The "Email Setup" button will open the email configuration screen. Email settings may be changed at any time. Check your mailbox to receive email messages, which include notifications of system status changes and periodic test status emails. Enter the interval at which emails will be received. For example, specifying an interval of 01:00:00 will send periodic emails every hour. Periodic emails are initiated with verification of the current status of the system, steps, pumps, and solenoid valves. In addition, system status change emails will be sent any time the system status changes, whether through automatic or manual operation. After all information has been properly entered, initiate a test email by clicking the "Test" button. If the settings are satisfactory and the test email has been received, press the "Save and Exit" button to save the changes and return to the main screen.

[0098] By clicking the "Start" button, the Select Recipe box appears. Open the most recent recipe text file by clicking the "Open XLS File" button. Recipe files are typically located in a folder on the desktop. Review the recipe to ensure the appropriate file is selected, and click the "Continue" button, which prompts for data entry information. Select the folder in which to save the data. The folder selected should reflect the bioreactor station used (4A, 4C, 5A, 5C). Select the frequency at which data points are saved (60 seconds). Once all settings have been entered, select the "Continue" button to start the process.

[0099] After selecting a recipe, the user will be asked to verify fluid and container settings as appropriate After selecting a recipe, the user will be prompted one last time before starting the automated decellularization process. Once started, the recipe can be paused by clicking the "Pause" button on the front panel. The user may need to pause the recipe execution for troubleshooting purposes. In the Pause state, the program will display a flashing "Resume" button, which prompts the user to continue execution after manual troubleshooting. In the Pause state, the user can switch valves, turn pumps on and off, and adjust pump settings. Although the pump can be run during the Pause state, the background color will still be gray, indicating an idle state. Data is continuously recorded even in the Pause state. The recipe resumes in the same step and with the same remaining step time as it was paused. If any pump settings are changed during the Pause, these changes will be retained after the Resume step. Pumps and solenoid valves can be manually set while the recipe is running. The user may need to manually adjust these settings for troubleshooting purposes, such as switching the perfusion pump to constant pressure instead of constant flow. The Override Control settings will be retained for the remainder of the step, as well as the Pause and Resume actions.

[0100] Once the recipe has completed the final step, the main status will display "Test Complete". Data Log File, which can be accessed through Microson Excel. Stores pressure-time stamped data during execution.

[0101] Although the above describes a specific preferred embodiment, it should be understood that the present invention is not limited thereto. It should be understood by those skilled in the art that various changes can be made to the disclosed embodiments, and these changes should fall within the scope of the present invention.

[0102] Unless otherwise specified, "a" or "an" means "one or more".

[0103] All publications, patent applications, and patents cited in this specification are herein incorporated by reference in their entirety.

Claims

1. An automated bioreactor system for decellularizing an organ, comprising: a main chamber configured to contain the organ, the main chamber having at least one reagent inlet, at least one reagent outlet, and at least one perfusion inlet; at least one reagent chamber containing a liquid reagent; at least one reagent conduit configured to deliver the liquid phase reagent from the at least one reagent chamber to the main chamber through the at least one reagent inlet; a reagent valve configured to control flow of a liquid-phase reagent through the at least one reagent conduit; at least one perfusion conduit configured to deliver the liquid phase reagent from the at least one reagent outlet through the at least one perfusion inlet to the organ contained in the main chamber; at least one perfusion pump configured to drive a flow of a liquid phase reagent through the at least one perfusion conduit; at least one perfusion pressure sensor configured to detect the pressure of a liquid agent flowing through the at least one perfusion conduit; a weight sensor configured to detect the weight contained in the main chamber; and A control system configured to: initiating a chamber filling process, which includes outputting a signal to open a valve associated with a reagent conduit to fill the main chamber with a predetermined amount of a liquid phase reagent; receiving an input of a weight detected by the weight sensor; outputting a signal based on the detected weight to control the reagent valve; receiving input representing a desired pressure of a liquid phase agent flowing through the at least one perfusion conduit during perfusion; After the chamber filling process is completed, the priming process begins, which includes: receiving input of pressure detected by the at least one perfusion pressure sensor; and outputting a signal to control the at least one perfusion pump to drive the liquid phase reagent flow based on the received input representing the desired pressure and the received input representing the detected pressure; waste chamber; a waste conduit configured to deliver waste fluid from the main chamber to the waste chamber; A purge fluid chamber configured to store a purge fluid; and A purge conduit is configured to direct purge fluid from the purge fluid chamber into the waste fluid.

2. The automated bioreactor system of claim 1, further comprising: A purge pump is configured to drive purge fluid from the purge fluid chamber through the purge conduit and into the waste fluid.

3. The automated bioreactor system of claim 2, wherein: The control system is configured to control the purge pump to drive the purge fluid from the purge chamber into the waste fluid based on input received of the weight detected by the weight sensor.

4. The automated bioreactor system of claim 2, wherein: The purge pump is also configured to drive the waste fluid from the main chamber to the waste chamber.

5. The automated bioreactor system of claim 1, further comprising: a drain conduit configured to deliver the mixture of waste fluid and purge fluid from the waste chamber to a drain; and A drain pump is configured to drive the mixture of the waste fluid and the purge fluid from the waste chamber through the drain conduit to a drain.

6. The automated bioreactor system of claim 1, further comprising: a plurality of secondary containment chambers, wherein the plurality of secondary containment chambers are configured to contain overflow of waste fluid from each of the main chamber, the waste chamber, and the purge fluid chamber; and A plurality of drain conduits, wherein the plurality of drain conduits are configured to deliver overflow of waste fluid contained in each of the plurality of secondary containment chambers from the plurality of secondary containment chambers to a drain.

7. The automated bioreactor system of claim 1, further comprising: A one-way drain valve is disposed along the waste conduit between the main chamber and the waste chamber to prevent waste fluid from flowing back into the main chamber.

8. The automated bioreactor system of claim 1, wherein: The control system calculates the volume of the liquid-phase reagent contained in the main chamber based on the detected weight and the density of the liquid-phase reagent.

9. The automated bioreactor system of claim 1, further comprising: at least one reagent valve configured to control flow of a liquid phase reagent through the at least one reagent conduit; Wherein the reagent conduit is flexible and the at least one reagent valve is a pinch valve that applies a force to the flexible reagent conduit to prevent the flow of the liquid phase reagent.

10. The automated bioreactor system of claim 1, further comprising: at least one temperature sensor configured to detect the temperature within the main chamber; and at least one temperature control system arranged to regulate the temperature within the main chamber, Wherein the control system is also configured to: receiving input of a temperature detected by the at least one temperature sensor; and A signal is output to control the temperature control system to adjust the temperature in the main chamber.

11. An organ perfusion system comprising: a main chamber configured to contain the organ and including a perfusion inlet for receiving a fluid for perfusing the organ; waste chamber; a waste conduit configured to deliver waste fluid from the main chamber to the waste chamber; A purge fluid chamber configured to store a purge fluid; and A purge conduit is configured to direct purge fluid from the purge fluid chamber into the waste fluid.

12. The organ perfusion system of claim 11, further comprising: A purge pump is configured to drive purge fluid from the purge fluid chamber through the purge conduit and into the waste fluid.

13. The organ perfusion system of claim 12, further comprising: a weight sensor configured to measure the weight of the main chamber; and The control system is configured to control the purge pump to drive the purge fluid from the purge chamber into the waste fluid based on the weight measured by the weight sensor.

14. The organ perfusion system according to claim 11, wherein: The purge pump is also configured to drive the waste fluid from the main chamber to the waste chamber.

15. The organ perfusion system of claim 11, further comprising: a drain conduit configured to deliver the mixture of waste fluid and purge fluid from the waste chamber to a drain; and A drain pump is configured to drive the mixture of the waste fluid and the purge fluid from the waste chamber through the drain conduit to a drain.

16. The organ perfusion system of claim 11, further comprising: a plurality of secondary containment chambers, wherein the plurality of secondary containment chambers are configured to contain overflow of waste fluid from each of the main chamber, the waste chamber, and the purge fluid chamber; and A plurality of drain conduits, wherein the plurality of drain conduits are configured to deliver overflow of waste fluid contained in each of the plurality of secondary containment chambers from the plurality of secondary containment chambers to a drain.

17. The organ perfusion system of claim 16, further comprising: A one-way drain valve is disposed along the waste conduit between the main chamber and the waste chamber to prevent waste fluid from flowing back into the main chamber.

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