CN107224612B - Automatic decellularization system and decellularization method - Google Patents

Automatic decellularization system and decellularization method Download PDF

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CN107224612B
CN107224612B CN201710401373.3A CN201710401373A CN107224612B CN 107224612 B CN107224612 B CN 107224612B CN 201710401373 A CN201710401373 A CN 201710401373A CN 107224612 B CN107224612 B CN 107224612B
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decellularization
container
reagent
liquid discharge
box body
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CN107224612A (en
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王红刚
韩卫东
米维
雷文诚
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Shaanxi Ruisheng Biotech Co ltd
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Shaanxi Ruisheng Biotech Co ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3629—Intestinal tissue, e.g. small intestinal submucosa
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3625—Vascular tissue, e.g. heart valves
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/005—Ingredients of undetermined constitution or reaction products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking

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Abstract

The invention relates to the technical field of decellularization, in particular to an automatic decellularization system and a decellularization method. Through automatic control, manual intervention in the process of removing cells from the biological material is reduced, and conditions are created for large-scale production and application of the biological material. An automated decellularization system comprising: the cell removing equipment comprises a cell removing container and a vibrating equipment, and the vibrating equipment is positioned below the cell removing container; a decellularization reagent supply device and a waste liquid discharge device which are respectively communicated with the decellularization container; and the control device is used for controlling the opening and closing of the oscillation device, controlling the supply of the decellularization reagent into the decellularization container by the decellularization reagent supply device, and controlling the liquid discharge device to discharge the liquid in the decellularization container.

Description

Automatic decellularization system and decellularization method
Technical Field
The invention relates to the technical field of decellularization, in particular to an automatic decellularization system and a decellularization method.
Background
In recent years, with intensive research on a cell removal technology of a biomaterial, a biomaterial substitute which has good biocompatibility, low immunogenicity, and a good repair effect after implantation is provided, wherein a three-dimensional structure of an original tissue is retained after cell removal of a biomaterial such as an animal membrane tissue of a porcine small intestine, a bovine pericardium, and the like, and a collagen protein with low immunogenicity is a main component, and becomes an optimal substitute for human tissue transplantation.
The existing cell removal technology is mainly completed in a reaction container through shaking table oscillation, when the biological material is produced on a large scale, manual intervention is more, so that the labor cost is higher, and the cell removal process needs harsh research and development and production conditions, so that pollution is easily caused when the manual intervention is more, the cell removal quality of the obtained biological material is difficult to guarantee, and the large-scale production and application of the biological material are limited.
Disclosure of Invention
The embodiment of the invention provides an automatic cell removing system and a cell removing method, which reduce manual intervention in a cell removing process of a biological material through automatic control and create conditions for large-scale production and application of the biological material.
In order to achieve the above purpose, the embodiment of the invention adopts the following technical scheme:
in one aspect, an embodiment of the present invention provides an automated decellularization system, including:
the cell removing equipment comprises a cell removing container and a vibrating equipment, and the vibrating equipment is positioned below the cell removing container;
a decellularization reagent supply device and a waste liquid discharge device which are respectively communicated with the decellularization container;
and the control device is used for controlling the opening and closing of the oscillation device, controlling the supply of the decellularization reagent into the decellularization container by the decellularization reagent supply device, and controlling the liquid discharge device to discharge the liquid in the decellularization container.
Optionally, the decellularization container comprises at least two decellularization grooves, and two adjacent decellularization grooves are communicated through a communication hole.
Optionally, the decellularization container comprises a first box body and a second box body, and the first box body is arranged in the second box body; a partition plate is arranged in the first box body and divides the first box body into at least two decellularization grooves, and the communication hole is formed in the partition plate between every two adjacent decellularization grooves;
the waste liquid discharge equipment comprises a liquid discharge port, a liquid discharge valve and a liquid discharge hole, wherein the liquid discharge port is arranged on the second box body and is communicated with the bottom space of the second box body; the drain valve is electrically connected with the control equipment.
Optionally, the number of the liquid discharge holes is at least two, and the liquid discharge holes are respectively formed in the bottom plate and the side plate of the first box body, and a cavity for communicating the liquid discharge port with the liquid discharge holes is formed by the bottom plate of the first box body and the bottom plate of the second box body in an enclosing manner.
Optionally, the decellularization container further comprises a first cover body which is matched and covered with the upper end face of the first box body, and a liquid inlet is formed in the first cover body;
the decellularization reagent providing equipment comprises a decellularization reagent storage tank and an infusion pump, wherein the input end of the infusion pump is communicated with the decellularization reagent storage tank, the output end of the infusion pump is communicated with the decellularization container through the liquid inlet, and the infusion pump is electrically connected with the control equipment.
Optionally, still be provided with the level gauge in the acellular container, controlgear with the level gauge electricity is connected, controlgear is used for controlling the level gauge is right detect the liquid level in the acellular container, and according to the liquid level signal that the level gauge sent is right the opening and closing of transfer pump are controlled.
In another aspect, an embodiment of the present invention provides an automated decellularization method applied to the above automated decellularization system, including:
placing the biological material in the decellularization container, and controlling the decellularization reagent providing device through the control device to supply a first preset amount of decellularization reagent into the decellularization container;
controlling an oscillation device to oscillate through the control device to perform decellularization on the biological material;
after the first period of time has elapsed, controlling a waste liquid discharge device by a control device to discharge the liquid in the decellularized container.
Optionally, the decellularization container comprises at least two decellularization grooves, and two adjacent decellularization grooves are communicated through a communication hole;
placing a biological material in the decellularized container comprises:
at least two biological materials are respectively placed in different acellular tanks.
Optionally, the decellularization container comprises a first box body and a second box body, and the first box body is arranged in the second box body; a partition plate is arranged in the first box body and divides the first box body into at least two decellularization grooves, and the communication hole is formed in the partition plate between every two adjacent decellularization grooves;
the waste liquid discharge equipment comprises a liquid discharge port, a liquid discharge valve and a liquid discharge hole, wherein the liquid discharge port is arranged on the second box body and is communicated with the bottom space of the second box body; the liquid discharge valve is electrically connected with the control equipment;
controlling the waste liquid discharge device to discharge the liquid in the decellularized container by the control device includes: and controlling the drainage valve to open by a control device, and discharging the liquid in the acellular container through the drainage hole and the drainage port.
Optionally, the decellularization container further comprises a first cover body which is matched and covered with the upper end face of the first box body, and a liquid inlet is formed in the first cover body; the decellularized reagent providing equipment comprises a decellularized reagent storage tank and an infusion pump, wherein the input end of the infusion pump is communicated with the decellularized reagent storage tank, the output end of the infusion pump is communicated with the decellularized container through the liquid inlet, and the infusion pump is electrically connected with the control equipment;
controlling the decellularization reagent supplying means to supply a first preset amount of the decellularization reagent into the decellularization vessel by the controlling means includes: and controlling the infusion pump to be switched on and off by the control equipment, and pumping a first preset amount of acellular reagent into the acellular container.
Optionally, a liquid level meter is further arranged in the acellular container, and the control device is electrically connected with the liquid level meter;
the control device controls the infusion pump to be switched on and off, and the step of pumping a first preset amount of acellular reagent into the acellular container specifically comprises the following steps: through controlgear control the level gauge is right the liquid level height of taking off cell reagent detects, if the level gauge detects the liquid level height of taking off cell reagent is less than preset height, then send first indicating signal for controlgear, controlgear basis first indicating signal control the transfer pump is opened, if the level gauge detects the liquid level height of taking off cell reagent reaches when presetting the height, send second indicating signal for controlgear, controlgear basis second indicating signal control the transfer pump is closed.
The embodiment of the invention provides an automatic decellularization system and a decellularization method, wherein a decellularization container, an oscillation device, a control device, a decellularization reagent supply device and a waste liquid discharge device which are communicated with the decellularization container are arranged, when the decellularization is carried out on a biological material, the biological material is placed in the decellularization container, the control device controls the decellularization reagent supply device to supply a decellularization reagent into the decellularization container, the control device controls the oscillation device to be started, the biological material can be decellularized under oscillation, after the decellularization is finished, the control device controls the waste liquid discharge device to discharge waste liquid generated by the decellularization, in the process, the automatic control can reduce manual intervention, reduce labor cost and reduce pollution in the decellularization process, improve the cell removal quality of the biological material and create conditions for the large-scale production and application of the biological material.
Drawings
FIG. 1 is a schematic structural diagram of an automated decellularization system according to an embodiment of the invention;
fig. 2 is a schematic structural diagram of a first box according to an embodiment of the present invention;
FIG. 3 is a schematic structural diagram of a decellularization vessel according to an embodiment of the invention;
FIG. 4 is a schematic structural diagram of a bottom of a first box according to an embodiment of the present invention;
fig. 5 is a schematic structural diagram of a first cover according to an embodiment of the present invention;
FIG. 6 is a schematic diagram of another automated decellularization system according to embodiments of the invention;
FIG. 7 is a schematic structural diagram of another automated decellularization system according to an embodiment of the invention;
fig. 8 is a schematic flow chart of an automated decellularization method according to an embodiment of the invention.
Detailed Description
An automated decellularization system and a decellularization method provided by the embodiments of the present invention are described in detail below with reference to the accompanying drawings.
In one aspect, the embodiments of the present invention provide an automated decellularization system, see fig. 1, including:
the cell removing device comprises a cell removing container 1 and a shaking device 2, wherein the shaking device 2 is positioned below the cell removing container 1;
a decellularization reagent supply device 3 and a waste liquid discharge device 4 which are respectively communicated with the decellularization container 1;
and the control device 5 is used for controlling the opening and closing of the oscillation device 2, controlling the supply of the decellularization reagent into the decellularization container 1 by the decellularization reagent supply device 3, and controlling the discharge of the liquid in the decellularization container 1 by the waste liquid discharge device 4.
The embodiment of the invention provides an automatic decellularization system, which is provided with a decellularization container 1, an oscillation device 2, a control device 5, a decellularization reagent supply device 3 and a waste liquid discharge device 4 which are communicated with the decellularization container 1, wherein when a biological material is decellularized, the biological material is placed in the decellularization container 1, the control device 5 controls the decellularization reagent supply device 3 to supply a decellularization reagent into the decellularization container 1, the control device 5 controls the oscillation device 2 to be opened, so that the biological material can be decellularized under oscillation, after the decellularization is completed, the control device 5 controls the waste liquid discharge device 4 to discharge waste liquid generated by the decellularization, in the process, the manual intervention can be reduced, and the labor cost can be reduced through automatic control, and can also reduce contamination during decellularization, improve the decellularization quality of the biomaterial
The control device 5 may comprise one or more processors, memories, user interfaces, network interfaces, and communication buses, among others.
The communication bus is used for controlling communication between the respective constituent elements in the apparatus 5. The user interface is used for plugging external equipment, such as a touch screen, a mouse, a keyboard and the like, so as to receive information input by a user. The network interface is used for the control device 5 to communicate with the outside, and mainly includes a wired interface and a wireless interface.
The memory may be used to store software programs and modules, databases, and program instructions/modules corresponding to the methods for decellularizing biological materials as described in the embodiments of the invention. The memory may include high speed random access memory, and may also include non-volatile memory, such as one or more magnetic storage devices, flash memory, or other non-volatile solid-state memory. In some examples, the memory may further include memory remotely located from the processor, and these remote memories may be connected to the control device via a network. Examples of such networks include, but are not limited to, the internet, intranets, local area networks, mobile communication networks, and combinations thereof.
The processor executes various functional applications and data processing by executing software programs and modules stored in the memory, for example, the processor calls a cell-removing application program of the biological material in the memory to realize a rapid and accurate cell-removing process of the biological material.
The specific structure of the decellularization vessel 1 is not limited, and the decellularization vessel 1 may be a large-sized reaction tank or a small-sized reaction tank.
In one embodiment of the present invention, referring to fig. 2, the decellularization container 1 includes at least two decellularization grooves 11, and two adjacent decellularization grooves 11 are communicated through a communication hole 6.
In the embodiment of the invention, by adopting the decellularization container 1 with the structure, when the cell of the biological material is removed, at least two biological materials can be respectively placed in different decellularization grooves 11, when the biological material is subjected to the cell removal under the oscillation, the biological materials can be prevented from being wound, so that the biological materials can be fully contacted with a decellularization reagent, the size of the decellularization grooves 11 is reasonably set, the biological materials can be conveniently flapped with the walls of the decellularization grooves 11 under the oscillation, the decellularization effect can be improved, compared with the use of one decellularization groove, the large-scale batch treatment can be realized, and after the decellularization is completed, the separation and the subsequent treatment are facilitated, so that the finished product quality of the decellularization of the biological materials can be improved. Further, since the two adjacent decellularization tanks 11 are communicated with each other through the communication hole 6, when the decellularization reagent is supplied into any one of the decellularization tanks 11 by the decellularization reagent supplying apparatus 3, the decellularization reagent can flow into the other decellularization tank 11 through the communication hole 6, so that the flow of the decellularization reagent during the decellularization process can be ensured, and a stable and uniform decellularization environment can be provided for the biological material in each of the decellularization tanks 11.
The specific structure of the decellularization container 1 is not limited, and the decellularization container 1 comprises at least two decellularization tanks 11, so that at least one partition plate can be arranged in the tank body or the tank body, and the tank body or the tank body is divided into the at least two decellularization tanks 11 by the partition plate; at least two reaction tanks can be spliced together to form the decellularization container 1.
In one embodiment of the present invention, referring to fig. 2, 3 and 4, the decellularized container 1 includes a first case 12 and a second case 13, the first case 12 is disposed in the second case 13; a partition plate 14 is arranged in the first box body 12, the partition plate 14 divides the first box body 12 into at least two decellularization grooves 11, and the communication hole 6 is formed in the partition plate 14 between every two adjacent decellularization grooves 11;
the waste liquid discharge apparatus 4 includes a liquid discharge port provided in the second casing 13 and communicating with the bottom space of the second casing 13, a liquid discharge valve a provided at the liquid discharge port, and a liquid discharge hole 7 provided in the first casing 12 and communicating with the bottom space of the second casing 13; the drain valve a is electrically connected to the control device 5.
In the embodiment of the present invention, the first casing 12 is disposed in the second casing 13, the first casing 12 is provided with the drain hole 7, and the bottom of the side wall of the second casing 14 is provided with the drain port, whereby the waste liquid after decellularization in the first casing 12 can be discharged through the drain hole 7 and the drain port when the drain valve a is opened. In the process, the fluid movement rule is complied, so that the labor can be saved, and the energy consumption caused by discharging the waste liquid through the waste liquid pump can be reduced.
The first housing 12 may have a circular or square horizontal cross-sectional shape, and the partition 14 may have an arc-shaped or straight plate as long as the first housing 12 can be divided into at least two decellularization cells 11, and accordingly, the decellularization cells 11 may have a circular or square horizontal cross-sectional shape depending on the shape of the first housing 12 and the partition 14.
In an embodiment of the present invention, the largest dimension of the decellularization tank 11 in the horizontal direction is less than or equal to 300 mm, and the height of the decellularization tank 11 in the vertical direction is greater than or equal to 100 mm. Can adapt to the oscillation amplitude of the oscillating shaking table 2, can provide enough movement space for at least three biological materials and prepare for the cell removal of the biological materials.
Illustratively, when the horizontal cross-sectional shape of the decellularization tank 11 is a square, the width of the decellularization tank 11 may be 200 mm, and the length may be 300 mm.
The arrangement of the decellularization grooves 11 may be various, and for example, the decellularization grooves 11 may be arranged in a row along the same linear array, or the decellularization grooves 11 may be arranged in a rectangular array. The whole structure of the decellularization tank 11 in a rectangular array is more compact than that in a linear array. Illustratively, as shown in FIG. 1, the partition 14 divides the space in the first housing 12 into 16 decellularization cells 11, and the 16 decellularization cells 11 are arranged in 4 rows and 4 columns. Of course, the partition 14 may also divide the space in the first housing 12 into 9 decellularization tanks 11, which is not limited herein.
It should be noted that, during the decellularization process, the decellularization reagent is usually a corrosive solvent, and therefore, the first tank 12 and the partition 14 may be made of a corrosion-resistant material, and preferably, the first tank 12 and the partition 14 are made of a stainless steel material. The stainless steel material is adopted, so that the strength of the acellular container 1 can be effectively improved, and acid-base corrosion can be prevented.
In practical applications, in order to avoid scratches on the biological material caused by the communication holes 6 and affect the quality of the finished product, it is preferable that the communication holes 6 are circular or elliptical.
In order to ensure the smooth circulation of the decellularization reagent and the effective blocking of the biological material, it is preferable that the diameter of the communication hole 6 is preferably 3 mm, so that the cell can be effectively blocked by various biological materials and the smooth circulation of the decellularization reagent can be ensured.
Preferably, referring to fig. 2, the communication holes 6 may be opened at the lower end of the partition 14, and the communication holes 6 are arranged in two rows in the vertical direction. Thus, the circulation of the decellularization reagent is facilitated, and the beating force of the partition plate 14 at the time of oscillation can be secured.
The drain hole 7 may be formed in a bottom plate or a side plate of the first casing 11. When the liquid discharge holes 7 are formed in the side plate, referring to fig. 4, the liquid discharge holes 7 are formed in the lower end of the side plate, and the two rows of the liquid discharge holes 7 are arranged in the vertical direction. This facilitates the flow of the cell-free waste liquid in each of the cell-free tanks 11.
Further, the drain hole 7 is circular or elliptical. Likewise, the circular or elliptical through holes can prevent the biological material from being scratched by the edges of the liquid discharge holes 7 or scratches during the oscillation process due to sharp corners of the liquid discharge holes 7, thereby causing defects that affect the quality of the finished biological material.
Preferably, the drain hole 7 has a diameter of 3 mm. Also can be convenient for blocking the biological material to lead the cell-free reagent to flow smoothly.
In another embodiment of the present invention, referring to fig. 4 and 5, the number of the drain holes 7 is at least two, and the drain holes are respectively opened on the bottom plate and the side plate of the first casing 12, and the bottom plate of the first casing 12 and the bottom plate of the second casing 13 enclose a cavity a for communicating the drain port and the drain holes 7. Like this, be favorable to the all-round discharge of outage on cell waste liquid through curb plate and the bottom plate to, through setting up cavity A, liquid in the first box 12 can pass through outage 7 is arranged to in the cavity A, then pass through the leakage fluid dram is discharged, can guarantee the smooth and easy discharge of waste liquid.
In order to completely discharge the waste liquid in the second tank 13 and reduce the residual, it is preferable that a bottom plate of the second tank 13 is disposed to be inclined, and a lowest portion of the bottom plate is disposed near the liquid discharge port.
In an embodiment of the present invention, the liquid discharge opening is further provided with a filter (not shown in the figure) for filtering the discharged waste liquid. After the completion of the decellularization, cell debris and fibrous impurities are dispersed in the decellularization reagent to form a decellularized waste liquid, and when the decellularized waste liquid is discharged through the liquid discharge port, the waste liquid is filtered by the filter, so that the clogging of the liquid discharge valve and the liquid discharge pump by the impurities can be prevented.
The first box 12 may be disposed in the second box 13 through a bracket, such that a bottom plate of the first box 12 and a bottom plate of the second box 13 enclose the cavity. A suspension structure may be provided in the second casing 13 to suspend the first casing 12 from the second casing 13, and the cavity a may be formed.
In another embodiment of the present invention, the side wall of the first box 12 is attached to the side wall of the second box 13. In this way, during the oscillation, the first casing 12 can be prevented from slipping with respect to the second casing 13, and the space inside the second casing 13 can be fully utilized.
In actual operation, if the decellularized container 1 is placed on the oscillating tray, slippage between the decellularized container 1 and the oscillating tray is likely to occur, which is disadvantageous to stable operation of the system.
In view of this, in an embodiment of the present invention, a connecting portion extends outwardly from a bottom edge of the second casing 13, and the connecting portion is detachably connected to the oscillating tray. Thus, the connecting part can be detached from the oscillation tray to replace the decellularization vessel 1 while preventing the decellularization vessel 1 from slipping with respect to the oscillation tray.
Here, the connection part and the oscillating tray may be in a snap connection or a screw connection.
It should be noted that the whole cell removing process is performed in a sterile environment, which can ensure the quality of the finished product of the biological material, and the first box 12 needs to be cleaned and sterilized after each cell removing process, so that, preferably, referring to fig. 3, the handle 8 is disposed on the upper end surface of the first box 12. In this way, when it is desired to sterilize the first casing 12, it can be lifted out of the second casing 13.
The decellularization reagent supply device 3 is not limited thereto, as long as it can supply a decellularization reagent into the decellularization vessel 1.
In an embodiment of the present invention, referring to fig. 6 and 7, the decellularization system further includes a first cover 15 that is matched and covered with the upper end surface of the first box 12, and a liquid inlet is disposed on the first cover 15;
the decellularization reagent providing equipment 3 comprises a decellularization reagent storage tank 31 and an infusion pump 32, wherein the input end of the infusion pump 32 is communicated with the decellularization reagent storage tank 31, the output end of the infusion pump is communicated with the decellularization container 1 through the liquid inlet, and the infusion pump 32 is electrically connected with the control equipment 5.
In the embodiment of the invention, the infusion pump 32 is used for injecting the decellularization reagent into the decellularization container 1, so that the decellularization reagent is convenient and quick, and the decellularization reagent can be prevented from splashing during injection and oscillation.
The output end of the transfer pump 32 may be connected to any one of the decellularization cells 11, so that the decellularization reagent may flow into each of the decellularization cells 11 through the communication hole 6 when the decellularization reagent is injected into the decellularization cell 11.
In order to prevent the biological materials in the different decellularization tanks 11 from being thrown against the upper end of the partition plate 14 during the oscillation process, referring to fig. 7, the first cover 15 has a flat plate-shaped structure, the upper edges of the side plate and the partition plate 14 are located in the same plane, and the first cover 15 covers the side plate and the partition plate 14.
Referring to fig. 6, a handle 9 may be further disposed on the upper surface of the first cover 15, and the handle 9 is grasped to open the first cover 15, which is convenient for operation.
Further, in order to facilitate observation of the decellularization process, a transparent observation window may be formed on the first cover body 15, and the transparent observation window may be made of a corrosion-resistant transparent material, or the first cover body 15 may be directly made of a corrosion-resistant transparent material.
In order to satisfy the requirement of decellularization, the second casing 14 may be made of stainless steel, and similarly, referring to fig. 3, a second cover 16 may be disposed at the opening of the second casing 14 to prevent the decellularization reagent from being contaminated.
The liquid level height of the decellularization reagent in the decellularization container 1 is not limited, and in practical application, the liquid level height can be set according to actual requirements, when the decellularization reagent is fed into the decellularization container 1 through the infusion pump 32, the feeding amount of the decellularization reagent can be calculated according to the set liquid level height, the running time of the infusion pump 32 can be calculated according to the feeding amount of the decellularization reagent and the running speed of the infusion pump 32, and the running of the infusion pump 32 can be automatically controlled.
In a preferred embodiment of the present invention, referring to fig. 7, a liquid level meter 10 is disposed in the decellularization container 1, the control device 5 is electrically connected to the liquid level meter 10, and the control device 5 is configured to control the liquid level meter 10 to detect the liquid level in the decellularization container 1 and control the liquid infusion pump 32 to be turned on or off according to a liquid level signal detected by the liquid level meter 10.
The liquid level meter 10 is arranged in the decellularization container 1, so that the liquid level in the decellularization container 1 is detected in real time, the liquid level height of a decellularization reagent in the decellularization container 1 can be controlled through the control device 5, and a proper decellularization reagent is provided for decellularization.
The level meter 10 may be disposed in any one of the decellularization tanks 11.
In another aspect, an embodiment of the present invention provides an automated decellularization method applied to the automated decellularization system described above, with reference to fig. 8, including:
step 1) placing a biological material in the decellularization container, and controlling the decellularization reagent providing device through a control device to supply a first preset amount of decellularization reagent into the decellularization container;
step 2) controlling an oscillation device to oscillate through the control device, and performing decellularization on the biological material;
and 3) after the first time, controlling a waste liquid discharge device to discharge the liquid in the acellular container through a control device.
The embodiment of the invention provides an automatic decellularization method, wherein a biological material is placed in a decellularization container, a decellularization reagent supply device is controlled by a control device to supply a decellularization reagent into the decellularization container, the control device is used for controlling an oscillation device to be started, so that the biological material can be decellularized under oscillation, after the decellularization is finished, a waste liquid discharge device is controlled by the control device to discharge waste liquid generated by the decellularization, in the process, manual intervention can be reduced, the labor cost can be reduced, pollution in the decellularization process can be reduced, the decellularization quality of the biological material can be improved, and conditions are created for large-scale production and application of the biological material.
The type of the biological material is not limited, and the biological material can be pig small intestine, sheep small intestine and the like.
In an embodiment of the present invention, the decellularization container includes at least two decellularization grooves, and two adjacent decellularization grooves are communicated through a communication hole;
placing a biological material in the decellularized container comprises:
at least two biological materials are respectively placed in different acellular tanks.
At least two of these means only spatial separation, and are not connected, and do not limit the total number of porcine small intestine or bovine pericardium. For example, one bovine pericardium may be trimmed into two biomaterials.
In the embodiment of the invention, the biological materials are separated by the partition board, the biological materials can be prevented from being intertwined, so that the biological materials can be fully contacted with the decellularization reagent, the biological materials can be fully flapped against the walls of the decellularization groove, the decellularization effect of the biological materials is improved, the processing scale of the biological materials can be increased, and in addition, after the decellularization is completed, the separation and subsequent operation are facilitated, so that the condition is created for large-scale production of the biological materials.
In a preferred embodiment of the present invention, the acellular container comprises a first box and a second box, wherein the first box is arranged in the second box; a partition plate is arranged in the first box body and divides the first box body into at least two decellularization grooves, and the communication hole is formed in the partition plate between every two adjacent decellularization grooves;
the waste liquid discharge equipment comprises a liquid discharge port, a liquid discharge valve and a liquid discharge hole, wherein the liquid discharge port is arranged on the second box body and is communicated with the bottom space of the second box body; the liquid discharge valve is electrically connected with the control equipment;
controlling the waste liquid discharge device to discharge the liquid in the decellularized container by the control device includes: and controlling the drainage valve to open by a control device, and discharging the liquid in the acellular container through the drainage hole and the drainage port.
The first box body is provided with the liquid discharge hole communicated with the bottom space of the second box body, the first box body is arranged in the second box body, the second box body is provided with the liquid discharge port communicated with the bottom space of the second box body, the liquid discharge valve is arranged at the liquid discharge port, the liquid discharge valve can be directly opened after cell removal is completed, waste liquid in each cell removal container can be discharged through the liquid discharge port, convenience and rapidness are achieved, and energy consumed by manpower and a waste liquid pump can be reduced.
In an embodiment of the present invention, the decellularization container further includes a first cover body that is matched and covered with the upper end surface of the first box body, and the first cover body is provided with a liquid inlet; the acellular reagent providing device comprises an acellular reagent storage tank and an infusion pump, the input end of the infusion pump is communicated with the acellular reagent storage tank, the output end of the infusion pump is communicated with the acellular container, and the infusion pump is electrically connected with the control device;
controlling the decellularization reagent supplying means to supply a first preset amount of the decellularization reagent into the decellularization vessel by the controlling means includes: and controlling the infusion pump to be switched on and off by the control equipment, and pumping a first preset amount of acellular reagent into the acellular container.
Wherein the operating time of the infusion pump may be calculated based on the first predetermined amount and the operating rate of the infusion pump, thereby enabling automatic feeding of a first predetermined amount of decellularization reagent into the decellularization vessel.
In the embodiment of the invention, the acellular reagent can be automatically injected into the acellular container by using the infusion pump, and the cover body can prevent the acellular reagent from splashing when being injected or oscillated.
In another embodiment of the present invention, a liquid level meter is further disposed in the acellular container, and the control device is electrically connected to the liquid level meter;
the control device controls the infusion pump to be switched on and off, and the step of pumping a first preset amount of acellular reagent into the acellular container specifically comprises the following steps: through controlgear control the level gauge is right the liquid level height of taking off cell reagent detects, if the level gauge detects when the liquid level height of taking off cell reagent is less than preset height, send first indicating signal for controlgear, controlgear basis first indicating signal control the transfer pump is opened, if the level gauge detects when the liquid level height of taking off cell reagent reaches preset height, send second indicating signal for controlgear, controlgear basis second indicating signal control the transfer pump is closed. Through setting up the level gauge, can carry out automated control to opening and closing of transfer pump to can be right the liquid level height of taking off cell reagent carries out effective control, improves the degree of automation of taking off the cell.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.

Claims (6)

1. An automated decellularization system, comprising:
the cell removing equipment comprises a cell removing container and a vibrating equipment, and the vibrating equipment is positioned below the cell removing container;
a decellularization reagent supply device and a waste liquid discharge device which are respectively communicated with the decellularization container;
the acellular container comprises a first box body and a second box body, and the first box body is arranged in the second box body; a partition plate is arranged in the first box body and divides the first box body into at least two cell removing grooves, a communication hole is formed in the partition plate between every two adjacent cell removing grooves, and the communication hole allows a cell removing reagent to pass through but does not allow biological materials to pass through;
the waste liquid discharge equipment comprises a liquid discharge port, a liquid discharge valve and a liquid discharge hole, wherein the liquid discharge port is arranged on the second box body and is communicated with the bottom space of the second box body; the bottom plate of each decellularization groove is provided with the liquid discharge hole, and the liquid discharge hole allows a decellularization reagent to pass through but does not allow biological materials to pass through;
the liquid discharge holes are at least two and are respectively arranged on the bottom plate and the side plate of the first box body, and the bottom plate of the first box body and the bottom plate of the second box body enclose a cavity for communicating the liquid discharge port and the liquid discharge holes;
and the control device is used for controlling the opening and closing of the oscillation device, controlling the supply of the decellularization reagent into the decellularization container by the decellularization reagent supply device, and controlling the opening and closing of the liquid discharge valve.
2. The automated decellularization system of claim 1,
the cell removing container also comprises a first cover body which is matched and covered with the upper end surface of the first box body, and a liquid inlet is formed in the first cover body;
the decellularization reagent providing equipment comprises a decellularization reagent storage tank and an infusion pump, wherein the input end of the infusion pump is communicated with the decellularization reagent storage tank, the output end of the infusion pump is communicated with the decellularization container through the liquid inlet, and the infusion pump is electrically connected with the control equipment.
3. The automated decellularization system of claim 2,
still be provided with the level gauge in the acellular container, controlgear with the level gauge electricity is connected, controlgear is used for controlling the level gauge is right the liquid level in the acellular container detects, and the basis is right the liquid level signal that the level gauge sent the opening and closing of transfer pump are controlled.
4. An automated decellularization method applied to the automated decellularization system according to any one of claims 1 to 3, comprising:
placing the biological material in the decellularization container, and controlling the decellularization reagent providing device through the control device to supply a first preset amount of decellularization reagent into the decellularization container;
controlling an oscillation device to oscillate through the control device to perform decellularization on the biological material;
after the first period of time has elapsed, the liquid in the decellularization vessel is discharged by controlling a liquid discharge valve by a control device.
5. The automated decellularization method of claim 4,
the cell removing container also comprises a first cover body which is matched and covered with the upper end surface of the first box body, and a liquid inlet is formed in the first cover body; the decellularized reagent providing equipment comprises a decellularized reagent storage tank and an infusion pump, wherein the input end of the infusion pump is communicated with the decellularized reagent storage tank, the output end of the infusion pump is communicated with the decellularized container through the liquid inlet, and the infusion pump is electrically connected with the control equipment;
controlling the decellularization reagent supplying means to supply a first preset amount of the decellularization reagent into the decellularization vessel by the controlling means includes: and controlling the infusion pump to be switched on and off by the control equipment, and pumping a first preset amount of acellular reagent into the acellular container.
6. The automated decellularization method of claim 5, wherein a liquid level meter is further disposed in the decellularization container, and the control device is electrically connected with the liquid level meter;
the control device controls the infusion pump to be switched on and off, and the step of pumping a first preset amount of acellular reagent into the acellular container specifically comprises the following steps: through controlgear control the level gauge is right the liquid level height of taking off cell reagent detects, if the level gauge detects the liquid level height of taking off cell reagent is less than preset height, then send first indicating signal for controlgear, controlgear basis first indicating signal control the transfer pump is opened, if the level gauge detects the liquid level height of taking off cell reagent reaches when presetting the height, send second indicating signal for controlgear, controlgear basis second indicating signal control the transfer pump is closed.
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