CN221117422U - Plant cell or tissue culture photoreactor - Google Patents

Abstract

The utility model belongs to the technical field of biological culture, and particularly relates to a plant cell or tissue culture photoreactor. The device comprises a light source device, a sealing head, a tank body, a ventilation device and an air supply system, wherein the bottom of the tank body is provided with a discharge hole, the ventilation device is arranged in the tank body and is positioned above the discharge hole, the air supply system is connected with the discharge hole, and the air supply system provides sufficient oxygen and proper carbon dioxide gas for culture solution in the tank body through the discharge hole and the ventilation device; the top of the tank body is sealed by the sealing head, the light source device is arranged on the plane of the sealing head, and the light source device provides illumination for plant tissues or cells. The aeration device reduces the transient shear force of the previous surface aeration type and bubbling air cylinder type on cells, increases the air content in the culture solution, improves the oxygen supply capacity, is beneficial to improving the air-liquid mass transfer rate of the nutrient components of the culture solution which are uniformly mixed, and ensures that plant cells or tissues obtain a proper growth environment.

Description

Plant cell or tissue culture photoreactor

Technical Field

The utility model belongs to the technical field of biological culture, and particularly relates to a plant cell or tissue culture photoreactor.

Background

In recent years, with the sustainable development of Chinese medicine industry, the application ways of the secondary metabolites of the medicinal plants are gradually increased, the medicinal value and the economic value are increased year by year, the market demand of the medicinal plants is also continuously increased, and the problems of the decrease of the wild resource accumulation of the medicinal plants and the like lead the medicinal plants to face serious contradiction between market supply and demand. Therefore, people develop artificially cultivated and domesticated wild varieties, and the medicinal materials with high part of medicinal value and special growth conditions face the problems of difficult cultivation, variety degradation, high cost and the like.

The plant suspension cell culture is a novel technology for rapidly obtaining mature plant cells and metabolites thereof, and the suspension cell culture technology is used for producing a large amount of medicinal components, thereby opening up a novel path for obtaining medicinal plant resources. The number of secondary metabolites separated from the cultured cells is more than six hundred, wherein the content of sixty medicinal plant metabolites exceeds or equals to that of the original plants, and more than two hundred plants are subjected to suspension cell culture, but the success of the suspension cell culture is small, the main reason is that the plant cells are uneven in thickness and extremely sensitive to shearing force although containing cellulose cell walls, and the plant cells or tissues are easy to adhere to form clusters, have high density and even have the characteristic of large viscosity in the later period of culture, so that the oxygen transfer rate and nutrient transfer are blocked, and the plant cell culture is more complex than a reactor for culturing microorganisms and animal cells due to the difference of culture conditions.

Meanwhile, dissolved oxygen is one of the most complex factors in the culture condition, the dissolved oxygen has a certain limit, and the cell growth and the product synthesis are influenced by the too high or the too low oxygen, especially in the high-density culture, the insufficient oxygen supply is often one of the main factors limiting the yield, so the regulation of dissolved oxygen parameters is critical to the plant cell culture.

The suspension culture of medicinal plant cells is suitable for large-scale industrial production. Bioreactors are important sites for carrying plant cells or tissues for physiological metabolism and for the synthesis of biological products. In addition to lack of illumination adjustment, the stirring rotation speed and the shape factor of the stirring paddle set by the device according to the original microorganism viscosity factor cause huge shearing force to influence proliferation of plant cells or tissues when the plant cells or tissues are used in the culture process. Later, researchers have developed an internal cross-flow and gas-mixing type airlift reactor, but cells or tissues which need illumination cannot be illuminated due to the restriction of the guide cylinder of the reactor, and plant cells or tissues are excessively clamped in the guide cylinder to form a dead volume, so that gas or nutrient components cannot be uniformly mixed in the culture process, and the growth and proliferation of the cells or tissues are affected. In recent years, a bubbling reactor has been developed, which removes the problem that the stirring paddle increases the ventilation amount, but the mixing property of the culture solution is insufficient because plant cells grow to the late cell tissue backlog, and if too much injected air suppresses the plant growth due to hydrodynamic stress, the large-scale production and popularization cannot be realized.

Disclosure of utility model

In view of the above problems, the present utility model aims to provide a plant cell or tissue culture photoreactor, which solves the problem that the existing plant cell or tissue culture reactor affects the proliferation of plant cells or tissues due to the large shearing force, insufficient illumination, insufficient dissolved oxygen and the like.

In order to achieve the above purpose, the present utility model adopts the following technical scheme:

The utility model provides a plant cell or tissue culture photoreactor, which comprises a light source device, a sealing head, a tank body, a ventilation device and a gas supply system, wherein the bottom of the tank body is provided with a discharge hole; the top of the tank body is sealed by the sealing head, and the light source device is arranged on the plane of the sealing head to provide illumination for plant cells or tissues in the culture solution.

The tank body is of an inverted frustum structure, and a culture solution parameter detection device is arranged on the side wall of the tank body.

The culture solution parameter detection device comprises a conductivity electrode, a T electrode, a pH electrode and an oxygen dissolving electrode, wherein the conductivity electrode is used for detecting the conductivity of the culture solution; the T electrode is used for detecting the temperature of the culture solution; the pH electrode is used for detecting the pH value of the culture solution; the dissolved oxygen electrode is used for detecting the dissolved oxygen degree of the culture solution.

The side wall of the tank body is provided with a sampling pipeline and a sampling valve arranged on the sampling pipeline, and the sampling valve is connected with a steam pipeline I.

The ventilation device comprises a ventilation column and a connecting plate connected to the lower end of the ventilation column, wherein the ventilation column is of a cylindrical structure with an opening at the bottom, and the side wall and the top wall of the ventilation column are of hollow structures; the connecting plate is of an annular structure and is used for being connected with the bottom of the tank body.

The ventilation column is made of a plurality of layers of stainless steel sintering nets, and the mesh number of the stainless steel sintering nets is gradually reduced from inside to outside.

The air supply system comprises a steam pipeline II, a compressed gas pipeline and a gas device, wherein one end of the compressed gas pipeline is connected with the discharge port, the other end of the compressed gas pipeline is connected with the gas device, the steam pipeline II is connected with the compressed gas pipeline through a shuttle valve, and the steam pipeline II provides high-temperature steam for the compressed gas pipeline to achieve the sterilization effect.

The light source device comprises a plurality of lamp posts arranged along the circumferential direction, the upper ends of the lamp posts are connected with the seal heads, and the lower ends of the lamp posts are inserted into the tank body.

The lamp post includes lamp area support, LED lamp area and safety cover, and wherein lamp area support is triangle-shaped columnar structure, and the upper end with the head is connected, and three sides of lamp area support all set up the LED lamp area, and the outside in LED lamp area is located to the safety cover, and upper end and lamp area leg joint.

A feeding port is arranged at the center of the end socket, and a plurality of feeding ports are arranged on the end socket along the circumferential direction; the lateral wall of the tank body is provided with an observation window.

The utility model has the advantages and beneficial effects that: the plant cell or tissue culture photoreactor provided by the utility model effectively solves the problem of insufficient oxygen supply when plant cells or tissues are cultured at high density. The combination of the ventilation device and the light source device can effectively reduce the shearing force of the gas on plant cells or tissues; simultaneously, the light source device can be used for providing a light source under ventilation, and the materials are better mixed, so that the large-scale culture of plant cells or tissues is promoted.

Drawings

FIG. 1 is a schematic diagram of a plant cell or tissue culture photoreactor according to the present utility model;

FIG. 2 is a top view of the tank of the present utility model;

FIG. 3 is a schematic view of the ventilation device according to the present utility model;

Fig. 4 is a schematic view of the structure of the lamp post according to the present utility model.

In the figure: 1-lamp post, 1 a-first lamp post, 1 b-second lamp post, 1 c-third lamp post, 101-lamp strip support, 102-LED lamp strip, 103-safety cover, 2-feed supplement mouth, 2 a-first feed supplement mouth, 2 b-second feed supplement mouth, 2 c-third feed supplement mouth, 3-head, 4-tank, 5-conductivity electrode, 6-T electrode, 7-steam pipeline I, 8-sampling pipeline, 9-discharge mouth, 10-observation window, 11-pH electrode, 12-ventilation device, 121-ventilation post, 122-connecting plate, 13-dissolved oxygen electrode, 14-compressed gas pipeline, 14-steam pipeline II, 15-compressed gas pipeline, 16-gas device, 17-feed port, 18-exhaust port.

Detailed Description

In order to make the objects, technical solutions and advantages of the present utility model more apparent, the present utility model will be described in detail with reference to the accompanying drawings and specific embodiments.

As shown in FIG. 1, the utility model provides a plant cell or tissue culture photoreactor, which comprises a light source device, a sealing head 3, a tank body 4, a ventilation device 12 and an air supply system, wherein a discharge port 9 is arranged at the bottom of the tank body 4, and a check valve is arranged at the discharge port 9. The ventilation device 12 is arranged in the tank body 4 and above the discharge hole 9, the air supply system is connected with the discharge hole 9, and the air supply system supplies sufficient oxygen and proper carbon dioxide gas to the culture solution in the tank body 4 through the discharge hole 9 and the ventilation device 12; the top of the tank body 4 is sealed by the sealing head 3, and the light source device is arranged on the sealing head 3 to provide illumination for plant cells or tissues in the culture solution.

In the embodiment of the utility model, the tank body 4 is of an inverted frustum structure, and a culture solution parameter detection device is arranged on the side wall of the tank body 4. The culture solution parameter detection device comprises a conductivity electrode 5, a T electrode 6, a pH electrode 11 and an oxygen dissolving electrode 13, wherein the conductivity electrode 5 is used for detecting the conductivity of the culture solution; the T electrode 6 is used for detecting the temperature of the culture solution; the pH electrode 11 is used for detecting the pH value of the culture solution; the dissolved oxygen electrode 13 is used to detect the dissolved oxygen degree of the culture solution. Specifically, the conductivity electrode 5 and the T electrode 6 are disposed on the left side of the tank body 4, the pH electrode 11 and the dissolved oxygen electrode 13 are disposed on the right side of the tank body 4, and each electrode is connected with the control system through an electrode connection line, and the control system can display numerical values.

Further, a sampling pipeline 8 and a sampling valve arranged on the sampling pipeline 8 are arranged on the side wall of the tank body 4, and the sampling valve is connected with the steam pipeline I7.

As shown in fig. 1 and fig. 3, in the embodiment of the utility model, the ventilation device 12 includes a ventilation column 121 and a connection plate 122 connected to the lower end of the ventilation column 121, wherein the ventilation column 121 has a cylindrical structure with an opening at the bottom, and the side wall and the top wall of the ventilation column 121 are hollow structures; the connection plate 122 has a ring-shaped structure and is detachably connected with the bottom of the tank body 4.

Specifically, the vent column 121 is made of a multi-layered stainless steel sintered mesh, and the mesh number of the multi-layered stainless steel sintered mesh gradually becomes smaller from inside to outside. The shape of the ventilation column 121 can be a cylinder or a cone, and the height and the diameter can be adjusted and changed according to the growth needs of plant tissues or cells and the volume of culture solution.

As shown in fig. 1 and 2, in the embodiment of the present utility model, the light source device includes a plurality of lamp posts 1 arranged along the circumferential direction, the upper ends of the lamp posts 1 are connected with the sealing heads 3, and the lower ends are inserted into the tank body 4.

As shown in fig. 1 and 4, in the embodiment of the utility model, a lamp post 1 includes a lamp strip support 101, an LED lamp strip 102 and a protection cover 103, wherein the lamp strip support 101 has a triangular column structure, the upper end is connected with a seal head 3, the LED lamp strip 102 is disposed on three sides of the lamp strip support 101, the protection cover 103 is covered on the outer side of the LED lamp strip 102, and the upper end is connected with the lamp strip support 101. Specifically, the protective cover 103 is a cylindrical high-temperature-resistant high-light-transmittance quartz glass cover, the lamp strip support 101 is a triangular iron column, the LED lamp strip 102 is attached to three sides, and the LED lamp strip 102 is externally connected with a light intensity regulator and a light cycle controller.

As shown in fig. 2, in the embodiment of the present utility model, a feeding port 17 is disposed at a central position of the sealing head 3, and a plurality of feeding ports 2 are also circumferentially disposed on the sealing head 3, where the feeding ports 2 and the lamp posts 1 are alternately arranged at intervals. A plurality of feed supplementing openings 2 are arranged, and a culture medium can be accessed and acid and alkali can be added for balancing. The sealing head 3 is also provided with an exhaust port 18, and the gas in the tank body 4 is discharged through the exhaust port 18.

In this embodiment, the feed inlet 17 of head 3 central point puts and is circular, lamp pole 1 and feed inlet 2 are three, and three lamp pole 1 is first lamp pole 1a, second lamp pole 1b and third lamp pole 1c respectively, and first lamp pole 1a, second lamp pole 1b and third lamp pole 1c are equilateral triangle overall arrangement, and inserts the inside of jar body 4 by the top of jar body 4, the effectual plant cell or the tissue illuminance demand that needs illumination that has improved, illumination distribution is even. Meanwhile, the layout of the lamp post 1 forms a baffle effect, and materials are better mixed. The three feed inlets 2 are a first feed inlet 2a, a second feed inlet 2b and a third feed inlet 2c, respectively, and the number and positions of the feed inlets 2 can be arranged as required.

Further, the side wall of the tank body 4 is also provided with an observation window 10. Specifically, the observation window 10 is a circular high temperature resistant and high perspective window, and is disposed at a position of half the height of the front surface of the tank 4.

As shown in fig. 1, in the embodiment of the present utility model, the air supply system includes a steam pipeline ii 14, a compressed air pipeline 15 and a gas device 16, wherein one end of the compressed air pipeline 15 is connected with the discharge port 9 by a screw valve, the other end is connected with the gas device 16, the steam pipeline ii 14 is connected with the compressed air pipeline 15 through a shuttle valve, and the steam pipeline ii 14 provides high-temperature steam for the compressed air pipeline 15, so as to realize the steam sterilization function.

Specifically, the gas device 16 adopts a three-stage gas filtering device, and the filtering effects of degreasing, dewatering and degerming of the gas entering the reactor are firstly achieved, so that the sterility of the gas is ensured on the external source. Next, the gas is filtered through the multi-layer stainless steel sintered net of the ventilation column 121, and the sterile state of the gas is further ensured before the gas enters the tank body 4 to be contacted with the culture solution or the cells. Different from the prior commercial reactors, the multi-level air sterilization device can effectively reduce the pollution in the reactor; secondly, the adjustable and selectable stainless steel sintering net is arranged in the reactor for filtering, so that the gas requirements of different plant cells or tissues and the shearing force resistance can be met, the phenomena of browning or slow proliferation of the plant cells or tissues in the amplifying culture process caused by the shearing force are effectively reduced, the better protection is improved, and the oxygen mass transfer coefficient in the culture solution is increased; and thirdly, the equilateral triangle light column arranged on the reactor head is introduced into the reactor, so that the illumination demand of plant cells or tissues needing illumination is effectively improved, illumination is uniformly distributed, and the aim of mixing culture solution in the culture process can be also fulfilled.

The invention solves the problems of uneven mixing and large shearing force of materials in the conventional plant reactor, adopts the detachable ventilation device 12 arranged at the inner bottom of the tank body 4, the ventilation column 121 is arranged at the central shaft position at the bottom of the tank body 4, the outside is connected with a sterile gas device, the ventilation column 121 is integrally manufactured by sintering stainless steel materials, the surface of a stainless steel sintering net is woven in a mat mode, the surface aperture precision is 1-2500 meshes, the pressure resistance is not easy to deform, and the ventilation column can be in a cylindrical shape or a conical shape. The vent column 121 may be selected in height and diameter. In this embodiment, the ventilation column 121 adopts a structure with more than three layers of stainless steel sintering nets, and each layer of stainless steel sintering net can have different pore diameters. The bottom of the ventilation column 121 is fixed by adopting a flange, so that the ventilation column 121 with different models can be conveniently detached and replaced. The breather 12 reduces the instantaneous shear force of the inlet air to the barrel, the circular inlet air tube at the bottom, and the flat air orifice plate and the shear force of the broken cells in the form of large bubbles in conventional designs.

In particular, plant tissue with a diameter of 3 mm or more proved to be suitable for ventilation of 0.06Nm

And plant tissue or cells with a diameter of less than 3mm are more suitable for aeration at 0.006Nm3/h. And can effectively protect the production of easily-oxidized polyphenol products.

In plant cell or tissue suspension culture, dissolved oxygen has a great influence on the growth and metabolism of plant cells or tissues. The invention effectively solves the problem of insufficient oxygen supply when plant cells or tissues are cultured in high density. The middle of the bottom of the reactor is provided with a gas column device, so that on one hand, the gas column type and the height can be changed according to the culture volume and the requirement of the species on the gas. On the other hand, the three-dimensional ventilation device can realize that gas can enter the culture solution in multiple directions from the outer surface of the column to the top of the column by utilizing the screen structure of the ventilation device, so that the gas is uniformly distributed, the diameter of bubbles is regulated by the design of the pore network of the ventilation column 121, the shearing force on cells or tissues generated after the input of the bubbles and during the crushing is reduced, the gas content in the culture solution is increased, the oxygen supply capability is improved, and the uniform mixing of nutrient components of the culture solution is facilitated. Meanwhile, the triangular design layout of the light column 1 is adopted to be introduced into the height of 1/3 position below the liquid level in the reactor, so that the illumination uniform intensity of plant cells or tissues below the liquid level can be effectively laid, the baffle plate function can be realized after ventilation in the plant cell or tissue culture process, vortex is formed, the mixing of culture solution is effectively assisted, and the proper growth environment of the plant cells or tissues is ensured.

The foregoing is merely an embodiment of the present utility model and is not intended to limit the scope of the present utility model. Any modification, equivalent replacement, improvement, expansion, etc. made within the spirit and principle of the present utility model are included in the protection scope of the present utility model.

Claims (10)

1. The plant cell or tissue culture photoreactor is characterized by comprising a light source device, a sealing head (3), a tank body (4), a ventilation device (12) and an air supply system, wherein a discharge port (9) is arranged at the bottom of the tank body (4), the ventilation device (12) is arranged in the tank body (4) and is positioned above the discharge port (9), the air supply system is connected with the discharge port (9), and the air supply system supplies oxygen to a culture solution in the tank body (4) through the discharge port (9) and the ventilation device (12); the top of the tank body (4) is sealed by the sealing head (3), and the light source device is arranged on the plane of the sealing head (3) to provide illumination for plant cells or tissues in the culture solution.

2. The plant cell or tissue culture photoreactor according to claim 1, characterized in that the tank (4) is of an inverted frustum structure, and the side wall of the tank (4) is provided with a culture fluid parameter detection device.

3. The plant cell or tissue culture photoreactor according to claim 2, characterized in that said culture fluid parameter detection means comprise a conductivity electrode (5), a T-electrode (6), a pH electrode (11) and an oxygen-dissolving electrode (13), wherein the conductivity electrode (5) is used for detecting the conductivity of the culture fluid; the T electrode (6) is used for detecting the temperature of the culture solution; the pH electrode (11) is used for detecting the pH value of the culture solution; the dissolved oxygen electrode (13) is used for detecting the dissolved oxygen degree of the culture solution.

4. The plant cell or tissue culture photoreactor according to claim 2, characterized in that the side wall of the tank (4) is provided with a sampling pipe (8) and a sampling valve arranged on the sampling pipe (8), and the sampling valve is connected with a steam pipe I (7).

5. The plant cell or tissue culture photoreactor according to claim 1, characterized in that the aeration device (12) comprises an aeration column (121) and a connecting plate (122) connected to the lower end of the aeration column (121), wherein the aeration column (121) has a cylindrical structure with an opening at the bottom, and the side wall and the top wall of the aeration column (121) have hollow structures; the connecting plate (122) is of an annular structure and is used for being connected with the bottom of the tank body (4).

6. The plant cell or tissue culture photoreactor according to claim 5, characterized in that the aeration column (121) is made of a multilayer stainless steel sintered mesh, the mesh number of which gradually decreases from inside to outside.

7. The plant cell or tissue culture photoreactor according to claim 1, characterized in that the gas supply system comprises a steam pipeline II (14), a compressed gas pipeline (15) and a gas device (16), wherein one end of the compressed gas pipeline (15) is connected with the discharge port (9), the other end is connected with the gas device (16), the steam pipeline II (14) is connected with the compressed gas pipeline (15) through a shuttle valve, and the steam pipeline II (14) provides high-temperature steam for the compressed gas pipeline (15) to realize the sterilization effect.

8. The plant cell or tissue culture photoreactor according to claim 1, characterized in that the light source device comprises a plurality of lampposts (1) arranged along the circumferential direction, the upper ends of the lampposts (1) are connected with the sealing heads (3), and the lower ends are inserted into the tank body (4).

9. The plant cell or tissue culture photoreactor according to claim 8, wherein the lamp post (1) comprises a lamp strip support (101), an LED lamp strip (102) and a protective cover (103), wherein the lamp strip support (101) is of a triangular columnar structure, the upper end of the lamp strip support is connected with the sealing head (3), the three sides of the lamp strip support (101) are provided with the LED lamp strip (102), the protective cover (103) is covered on the outer side of the LED lamp strip (102), and the upper end of the protective cover is connected with the lamp strip support (101).

10. The plant cell or tissue culture photoreactor according to claim 1, characterized in that a feeding port (17) is arranged at the center of the sealing head (3), and a plurality of feeding ports (2) are arranged on the sealing head (3) along the circumferential direction; the side wall of the tank body (4) is provided with an observation window (10).