CN110713928A

CN110713928A - Sleeve type negative pressure biological reaction device and method

Info

Publication number: CN110713928A
Application number: CN201911111190.3A
Authority: CN
Inventors: 周广东; 白宝帅; 张沛灵; 贾丛励; 刘豫; 曹谊林
Original assignee: Weifang Medical University
Current assignee: Weifang Medical University
Priority date: 2019-11-14
Filing date: 2019-11-14
Publication date: 2020-01-21

Abstract

The invention discloses a sleeve type negative pressure biological reaction device which structurally comprises a culture tank body, wherein a tank cover is arranged on the culture tank body, a rubber diaphragm is tightly connected between the culture tank cover and the culture tank body, a support rod is arranged in the middle of the culture tank body, a sleeve is movably arranged on the support rod, the sleeve is connected with a first ejector rod arranged at an opening at one side of the culture tank body, the first ejector rod is connected with a second ejector rod through a connecting piece, a sealing gasket is arranged at the part of the second ejector rod, which extends into the wall of the culture tank body, the tail end of the second ejector rod is connected with a pressurizing assembly, the pressurizing assembly acts on the tail end of the second ejector rod, acts on the first ejector rod through force transmission, and jacks; the invention has simple structure, can well simulate the negative pressure state of the human joint cavity, and the negative pressure reaches the negative pressure of the human joint cavity, and is fixedly placed into a bioreactor for culture every day when the cartilage is cultured, thereby better simulating the real environment in the body and better promoting the formation of the cartilage.

Description

Sleeve type negative pressure biological reaction device and method

Technical Field

The invention relates to the field of biochemical test reaction vessels, in particular to a sleeve type negative pressure biological reaction device and a method.

Background

A bioreactor is a reaction system which utilizes naturally existing microorganisms or microorganisms with special degradation capacity to inoculate liquid phase or solid phase, is also used for experimental tools for culturing biological cell tissues, provides 1-hour high-pressure culture environment for the cell tissues every day, and has the advantages that strict aseptic conditions for culturing the cell tissues are required, once the polluted cell tissues are subjected to irrecoverable lethal shock, the asepsis is the basis of cell culture, the sufficient nutrient environment (including supply of nutrients (the degradation rate of biological factors required by various cells is high) and removal of waste) is realized, and the liquid is changed at regular intervals for meeting the requirements of the biological tissues.

Patent 2018112940730 discloses a biological culture reaction device, which comprises a culture chamber and a pressurizing assembly, wherein the pressurizing assembly comprises an inner pressure head, an upper pressure plate, a lower pressure plate and a pressure spring; the upper end face of the inner pressing head is connected with an upper pressing plate through a bolt, the lower end of the inner pressing head is sleeved with a lower pressing plate, the lower part of the inner pressing head is provided with a boss, the upper end of a through hole of the lower pressing plate is provided with a convex ring, and the lower pressing plate is matched with the boss at the lower part of the inner pressing head through the convex ring to be axially positioned, so that the lower pressing plate can move upwards along with the inner pressing; the inner pressure head is sleeved with a pressure spring, and two ends of the pressure spring are respectively in contact connection with the upper pressure plate and the lower pressure plate. The inner pressure head and the lower pressure plate are respectively in pressure contact connection with the silica gel membrane in the culture cabin, wherein the lower pressure plate is used for tightly pressing the silica gel membrane and the upper end face of the culture tank body to enable the silica gel membrane and the upper end face of the culture tank body to be tightly sealed, and the inner pressure head applies set pressure to liquid in the culture tank body through the silica gel membrane. The main technical problem solved by the technology of the patent is that the sealing condition is good, and the pollution of the reactor is reduced. But it is not very good to simulate the physical environment that cartilage in the joint cavity is subjected to under the environment of positive pressure alone.

At present, research shows that negative pressure can promote cell growth, but no negative pressure bioreactor exists in the market, and human joints and other parts exist under negative pressure for a long time, so the negative pressure bioreactor is also a bionic technology.

Disclosure of Invention

The invention mainly solves the technical problem of how to provide a sleeve type negative pressure biological reaction device and a method, which can simulate the conditions of joint motion and resting state negative pressure in a joint cavity, better store cartilage formation and promote the directional differentiation of cartilage.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows:

a sleeve type negative pressure biological reaction device is characterized in that: the structure comprises a culture tank body, wherein a tank cover is arranged on the culture tank body, a rubber diaphragm is tightly connected between the culture tank cover and the culture tank body, and a base plate is arranged between the culture tank cover and the rubber diaphragm, so that the stress is uniform during sealing; in order to increase the stability of the culture tank during placement, a fixed small arm is arranged at the bottom of the culture tank body and is clamped with a fixed notch to rotate for about 5 degrees and then screwed down to be fixed with the culture tank body; the middle of the culture tank body is provided with a support rod, a sleeve is movably mounted on the support rod, a first ejector rod is arranged at an opening on one side of the sleeve and the culture tank body and connected with a second ejector rod through a connecting piece, a sealing gasket is arranged on a part, extending into the wall of the culture tank body, of the second ejector rod, the tail end of the second ejector rod is connected with a pressurizing assembly, the pressurizing assembly acts on the tail end of the second ejector rod, force is transmitted to the first ejector rod, the sleeve is jacked up to form a rubber diaphragm through the first ejector rod, and corresponding negative pressure.

Preferably, the bottom of the culture tank body is screwed and connected with the pressure sensor through a threaded opening.

Preferably, the pressurizing sleeve is connected with the air cylinder, the oil cylinder and the servo motor or the linear motor; when the device works, the sleeve is jacked up by the first ejector rod to generate corresponding negative pressure, after negative pressure stimulation is completed, the pressurizing assembly relaxes, and the sleeve and the ejector rod return to the initial state.

A sleeve type negative pressure biological reaction method comprises the following steps:

step 1: putting the culture into the culture tank body, adding corresponding culture solution, covering a matched culture dish cover to allow oxygen and carbon dioxide to enter and prevent pollution, putting the culture dish into a cell culture box for normal culture according to the principle of the original culture dish;

step 2: carrying out negative pressure stimulation within a fixed time every day, taking out the culture tank body from the incubator before carrying out the negative pressure stimulation, taking down the culture dish cover, covering a sterilized silica gel membrane and covering the culture tank cover, and then screwing down by using a coaxial wrench matched with the culture tank cover;

and step 3: then putting the culture tank into a pressurizing reaction box, inserting the two tail ends of the ejector rods into preset positions in a pressurizing assembly, and fixing the culture tank;

and 4, step 4: opening the programmed pressurizing device, pressurizing the ejector rod II through the pressurizing assembly to drive the ejector rod I and the sleeve to move upwards, ejecting the silica gel membrane on the sleeve through the lever principle to generate negative pressure, then loosening the pressurizing device, and repeating the steps;

and 5: and after the negative pressure stimulation is completed, taking down the culture tank body, unscrewing the culture tank cover, taking down the tank cover and the diaphragm in a fume hood, replacing the culture dish cover, putting the culture dish cover into the cell culture box, and performing conventional culture again to complete the negative pressure biological reaction.

Preferably, the temperature in the pressurized reaction tank in the step 3 is set to be constant at 37 ℃.

The invention has the beneficial effects that:

the invention has simple structure, can well simulate the negative pressure state of the human joint cavity, and the negative pressure reaches-0.17-0.6 bar of the negative pressure of the human joint cavity, and is fixedly placed into a bioreactor for culture every day when the cartilage is cultured, thereby better simulating the real environment in the body and better promoting the formation of the cartilage.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without inventive efforts, wherein:

FIG. 1 is a schematic view of a reactor and a pressure jacket connection according to the present invention;

reference numerals:

1-a culture tank body; 2-culture pot cover; 3-a rubber diaphragm; 4-fixing the forearm; 5-fixing the notch; 6-supporting rods; 7-a sleeve; 8-a first ejector rod; 9-ejector rod II; 10-a seal gasket; 11-pressure sensor.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1, in an embodiment of the present invention, a sleeve-type negative pressure bioreactor is characterized in that: the structure comprises a culture tank body 1, wherein a tank cover 2 is arranged on the culture tank body 1, a rubber diaphragm 3 is tightly connected between the culture tank cover 2 and the culture tank body 1, and a base plate is arranged between the culture tank cover 2 and the rubber diaphragm 3, so that the stress is uniform during sealing; in order to increase the stability of the culture tank during placement, the bottom of the culture tank body 1 is provided with a small fixed arm 4, and the small fixed arm 4 is clamped with a fixed notch 5 and is rotated by about 5 degrees to be screwed down so as to be fixed with the culture tank body; the middle of the culture tank body 1 is internally provided with a support rod 6, the support rod is movably provided with a sleeve 7, the sleeve 7 is connected with a first ejector rod 8 arranged at an opening at one side of the culture tank body 1, the first ejector rod 8 is connected with a second ejector rod 9 through a connecting piece, a sealing gasket 10 is arranged at a part of the second ejector rod 9 extending into the wall of the culture tank body 1, the tail end of the second ejector rod 9 is connected with a pressurizing assembly, external force is applied to the first ejector rod 8 through the lever principle, the sleeve 7 jacks up the rubber diaphragm 3 through the first ejector rod 8, and corresponding negative.

In the specific implementation process, the bottom of the culture tank body 1 is screwed with the pressure sensor 11 through a threaded opening.

In the specific implementation process, the pressurizing sleeve is connected with the air cylinder, the oil cylinder and the servo motor or the linear motor; under normal conditions, the upper end of the sleeve is flush with the bottom surface of the silicon membrane, and when the device works, external force is applied to the first 8 ejector rods according to the lever principle, the sleeve 7 jacks up the rubber membrane 3 through the first 8 ejector rods, and corresponding negative pressure is generated.

In a specific implementation process, the temperature in the pressurized reaction box in the step 3 is set to be constant at 37 ℃.

The invention has the beneficial effects that:

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.

Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.

Claims

1. A sleeve type negative pressure biological reaction device is characterized in that: the structure comprises a culture tank body, wherein a tank cover is arranged on the culture tank body, a rubber diaphragm is tightly connected between the culture tank cover and the culture tank body, and a base plate is arranged between the culture tank cover and the rubber diaphragm, so that the stress is uniform during sealing; in order to increase the stability of the culture tank during placement, a fixed small arm is arranged at the bottom of the culture tank body and is clamped with a fixed notch to rotate for about 5 degrees and then screwed down to be fixed with the culture tank body; a supporting rod is arranged in the middle of the inside of the culture tank body, a sleeve is movably mounted on the supporting rod, the sleeve is connected with a first ejector rod at an opening on one side of the culture tank body, the first ejector rod is connected with a second ejector rod through a connecting piece, a sealing gasket is arranged on a part, extending into the wall of the culture tank body, of the second ejector rod, the tail end of the second ejector rod is connected with a pressurizing assembly, the pressurizing assembly acts on the tail end of the second ejector rod, force is transmitted to the first ejector rod, the sleeve is jacked up to a rubber diaphragm through the first;

the biological reaction method comprises the following steps:

and 5: after the negative pressure stimulation is completed, the culture tank body is taken down, the culture tank cover is unscrewed, then the tank cover and the diaphragm are taken down in a fume hood, the culture dish cover is replaced, the cell culture box is placed in the culture tank for conventional culture again, and the negative pressure biological reaction is completed.

2. The telescopic negative pressure bioreactor apparatus according to claim 1, wherein: the bottom of the culture tank body is screwed up and connected with the pressure sensor through a threaded opening.

3. The telescopic negative pressure bioreactor apparatus according to claim 1, wherein: the pressurizing sleeve is connected with the air cylinder, the oil cylinder and the servo motor or the linear motor; when the device works, the sleeve is jacked up by the first ejector rod to generate corresponding negative pressure, after negative pressure stimulation is completed, the pressurizing assembly relaxes, and the sleeve and the ejector rod return to the initial state.

4. The telescopic negative pressure bioreactor apparatus according to claim 1, wherein: and 3, setting the temperature in the pressurized reaction box to be constant at 37 ℃.