Airlift bioreactor for amplification culture of taxus cuspidata stem cells

Abstract

The utility model discloses an airlift bioreactor for amplification culture of taxus cuspidata stem cells. The airlift bioreactor comprises a tank body, a rotatable spiral guide cylinder and a microbubble generator, the microbubble generator is arranged in the middle of the bottom of the tank body, and the rotatable spiral guide cylinder is arranged in the tank body and fixed above the microbubble generator. According to the utility model, the micro-bubble generator is arranged at the air vent at the bottom of the bioreactor tank body, the rotatable spiral baffle is designed in the guide cylinder, and the spiral baffle in the guide cylinder is driven to rotate by utilizing kinetic energy generated by uniform rising of micro-bubbles through a micro-bubble group generated by the micro-bubble generator, so that fluid forms vortex flow in a spiral channel; and meanwhile, the collision probability of the microbubbles is reduced under the action of centrifugal force. Through a non-mechanically driven dynamic mixing mode, the oxygen transfer and mass transfer bottleneck in a high-viscosity culture system is effectively solved, and efficient reaction equipment is provided for large-scale culture of plant cells.

Description

Technical Field

[0001] This invention belongs to the field of bioreactor technology, specifically relating to an airlift bioreactor, which is particularly suitable for large-scale culture of medicinal plant stem cells, especially for the synergistic optimization of oxygen transfer efficiency and mass transfer performance during the suspension culture of Taxus chinensis stem cells. Background Technology

[0002] Paclitaxel, a diterpenoid compound found in yew trees, has the ability to interfere with the normal breakdown of microtubules during cell division, effectively inducing apoptosis in cancer cells. To date, paclitaxel is widely recognized as one of the most effective anticancer drugs and is widely used in the treatment of various cancers such as ovarian cancer, breast cancer, and lung cancer. Utilizing yew tree suspension cell culture technology is one of the effective ways to obtain paclitaxel. This method features rapid cell proliferation, is not limited by season, allows for large-scale culture, and can provide a large quantity of homogeneous plant cell cultures.

[0003] Northeast yew stem cells are undifferentiated cells located in the cambium meristem. They possess characteristics such as rapid cell proliferation, thin cell walls, numerous small vacuoles, and fast growth, overcoming the limitations of limited division capacity and unstable genetic traits in traditional Northeast yew callus dedifferentiated cell culture. Utilizing plant stem cell large-scale culture technology to produce high-value secondary metabolites has become an important technological pathway to overcome the bottleneck of natural product resources.

[0004] To achieve the leap from laboratory research to industrial application of medicinal plant stem cell culture technology, multi-stage scale-up culture using bioreactor systems is essential. An airlift bioreactor is a bioreactor that uses gas as a power source, driving the circulation of the culture medium through the upward flow of gas to achieve mixing, mass transfer, and heat transfer. Its core feature is the absence of mechanical stirring devices; it relies on the density difference between the gas and liquid phases to form a circulating flow, avoiding cell damage caused by mechanical stirring. However, conventional airlift reactors produce large-diameter bubbles with small gas-liquid contact areas and low oxygen transfer coefficients, making it difficult to meet the dissolved oxygen requirements of high-density cell culture. Furthermore, traditional airlift reactors use vertical guide tubes, resulting in a single liquid circulation path. The laminar flow region leads to uneven distribution of nutrients and metabolites, especially in viscous culture media, easily forming mass transfer dead zones. While mechanically stirred bioreactors can enhance mass transfer, the high shear forces can damage sensitive cells, and the complex mechanical structure increases the risk of bacterial contamination.

[0005] In summary, the rapid growth, high proliferation rate, high shear tolerance, and large consumption of nutrients and oxygen of yew stem cells significantly limit stem cell proliferation and paclitaxel production due to the oxygen and mass transfer rates of the culture medium in traditional bioreactors. Therefore, the airlift bioreactor required for culturing Northeast yew stem cells must not only meet the requirements of good mixing performance and low shear force of the culture medium, but also maintain a high oxygen solubility. Summary of the Invention

[0006] This invention addresses the problems of low mass and oxygen transfer efficiency and uneven mixing in bioreactors. It provides an airlift bioreactor for the scale-up culture of *Taxus chinensis* stem cells, employing an innovative reactor design based on the rotation of a spiral baffle within a microbubble-driven flow guide tube. The core of this design involves placing a microbubble generator at the bottom vent of the bioreactor tank and incorporating a rotatable spiral baffle within the flow guide tube. The microbubble clusters generated by the microbubble generator significantly expand the gas-liquid contact area. The kinetic energy of the uniformly rising microbubbles drives the rotation of the spiral baffle within the flow guide tube. The specific spiral and structural design of the spiral baffle creates a vortex flow within the spiral channel, enhancing gas-liquid mixing and mass diffusion, and significantly extending the gas-liquid mixing path. Simultaneously, the centrifugal force of the microbubbles reduces the probability of collisions, thereby reducing the aggregation of microbubbles caused by bottom pressure due to the high reactor diameter-to-height ratio, and enhancing the uniformity of microbubble distribution in the culture medium. This invention effectively solves the oxygen and mass transfer bottlenecks in high-viscosity culture systems through a non-mechanically driven dynamic mixing mode, providing a highly efficient reaction device for large-scale plant cell culture.

[0007] The objective of this utility model is achieved through the following technical solution:

[0008] An airlift bioreactor for amplified culture of Taxus chinensis stem cells includes a tank, a rotatable spiral guide tube, and a microbubble generator. The microbubble generator is located at the bottom center of the tank, and the rotatable spiral guide tube is located inside the tank and fixed above the microbubble generator. The top cover of the tank is provided with an inlet, an outlet, and an exhaust port. The culture medium enters the tank through the inlet, and sterile air is dispersed into microbubbles by the microbubble generator and enters the tank. The kinetic energy of the microbubble airflow promotes the culture medium to spiral upward inside the rotatable spiral guide tube and then descend outside the rotatable spiral guide tube to achieve circulation.

[0009] Furthermore, the rotatable spiral guide tube includes a spiral baffle and a guide tube. The guide tube is fixed in the center inside the tank, and the upper and lower ends of the spiral baffle are rotatably and sealingly connected to the top cover of the tank and the microbubble generator, respectively.

[0010] Furthermore, the ratio of the pitch of the spiral baffle to the height of the guide tube is 1:1, the ratio of the spiral diameter to the diameter of the guide tube is 4:5, and the ratio of the diameter of the guide tube 2 to the diameter of the microbubble generator is 4:3.

[0011] Furthermore, the spiral baffle includes a gradient spiral baffle body, the bottom of which is embedded in the center of the microbubble generator via a sealed bearing, and the top of which is mounted on the top cover of the tank via a sealed bearing.

[0012] Furthermore, the airlift bioreactor also includes a temperature control jacket surrounding the outer perimeter of the tank.

[0013] Furthermore, the temperature control jacket is connected to the constant temperature water tank via a circulating water pump.

[0014] Furthermore, the airlift bioreactor also includes a temperature probe, a dissolved oxygen electrode, and a pH electrode; the temperature probe, dissolved oxygen electrode, and pH electrode are respectively installed inside the tank through the temperature probe port, dissolved oxygen electrode port, and pH electrode port provided on the top cover of the tank.

[0015] Furthermore, the airlift bioreactor also includes a precursor feed bottle, an inducer feed bottle, a feedback inhibitor feed bottle, and an acid / base feed tank. The top cover of the tank is respectively provided with an acid / base feed port, a precursor / inducer feed port, and a feedback inhibitor feed port. The precursor feed bottle is connected to the precursor / inducer feed port through a precursor feed peristaltic pump, the inducer feed bottle is connected to the precursor / inducer feed port through an inducer feed peristaltic pump, the feedback inhibitor feed bottle is connected to the feedback inhibitor feed port through a feedback inhibitor feed peristaltic pump, and the acid / base feed tank is connected to the acid / base feed port through an acid / base feed peristaltic pump.

[0016] Furthermore, the microbubble generator is connected to an air supply device, which includes a check valve, a flow rotor, an intake air filter, and an air pump. The air pump filters compressed air through the intake air filter into sterile air. The sterile air is then regulated by the flow rotor and enters the microbubble generator through the check valve.

[0017] This utility model has the following beneficial effects:

[0018] This invention significantly improves the oxygen transfer and mixing performance of an airlift bioreactor through the synergistic design of a microbubble generator and a spiral guide tube. The microbubble clusters generated by the microbubble generator increase the gas-liquid contact area and improve oxygen transfer efficiency; simultaneously, the microbubble airflow drives the spiral baffle to rotate, creating a vortex flow that enhances the gas-liquid contact time, while centrifugal force inhibits bubble coalescence, thus solving the mass transfer dead zone problem in high-viscosity culture media. This invention employs a pneumatic circulation mode without mechanical stirring, reducing shear force while maintaining system sealing and effectively minimizing the risk of contamination. Furthermore, the integrated temperature control, feeding, and online monitoring system allows for precise adjustment of culture parameters based on metabolic kinetics. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of this utility model, the accompanying drawings used in the description of the embodiments of this utility model will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on the content of the embodiments of this utility model and these drawings without creative effort.

[0020] Figure 1 This is a schematic diagram of the main structure of an airlift bioreactor for amplified culture of Taxus chinensis stem cells as described in Embodiment 1 of this utility model;

[0021] Figure 2 This is a schematic diagram of the rotatable spiral guide tube structure described in Embodiment 1 of this utility model;

[0022] Figure 3 This is a schematic diagram of the spiral baffle structure described in Embodiment 1 of this utility model;

[0023] Figure 4 This is a schematic diagram illustrating the composition and principle of an improved airlift bioreactor for amplified culture of Taxus chinensis stem cells as described in Embodiment 1 of this utility model.

[0024] In the picture:

[0025] 1-Tank body; 2-Temperature control jacket; 3-Rotable spiral guide tube; 31-Spiral baffle; 32-Guide tube; 4-Microbubble generator; 5-Check valve; 6-Flow rotor; 7-Inlet air filter; 8-Air pump; 9-Exhaust air filter; 10-Dissolved oxygen electrode; 11-pH electrode; 12-Temperature probe; 13-Constant temperature water tank; 14-Circulating water pump; 15-Precursor feed bottle; 16-Inducer feed bottle; 17-Feedback inhibitor feed bottle; 18-Precursor feed peristaltic pump; 19-Inducer feed peristaltic pump; 20-Feedback inhibitor feed peristaltic pump; 21-Acid / alkali feed peristaltic pump; 22-Acid / alkali feed tank; 23-Three-way solenoid valve; 24-Piston pump; 25-Sampling bottle. Detailed Implementation

[0026] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, the accompanying drawings show only the parts relevant to the present invention, not the entire structure.

[0027] In the description of this utility model, unless otherwise expressly specified and limited, the terms "connected," "linked," and "fixed" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part of the device. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.

[0028] Example 1:

[0029] like Figure 1 This embodiment describes an airlift bioreactor for the scale-up culture of Taxus chinensis stem cells, comprising a tank 1, a rotatable spiral guide tube 3, and a microbubble generator 4. The microbubble generator 4 is located at the bottom center of the tank 1, and the rotatable spiral guide tube 3 is located inside the tank 1 and fixed above the microbubble generator. The top cover of the tank 1 is provided with an inlet, an outlet, and an exhaust port. The inlet is used to add Taxus chinensis stem cells and culture medium to the tank during initial culture, and the outlet is connected to a sampling system for quantitative sampling and detection of cell biomass and effective component content during the culture process. The culture medium containing Taxus chinensis stem cells enters the tank through the inlet. Sterile air is dispersed into microbubbles by a microbubble generator and enters the tank. The kinetic energy of the microbubble airflow promotes the Taxus chinensis stem cell culture medium to spiral upward in the rotatable spiral guide tube 3. The disturbance process in the rotatable spiral guide tube 3 further promotes mass transfer and oxygen transfer efficiency, and the medium circulates by descending outside the guide tube. Therefore, a complex mechanical stirring system is not required, which reduces the energy consumption of the system and ensures the airtightness of the system. It reduces the risk of bacterial contamination and avoids cell damage caused by mechanical stirring.

[0030] Preferably, the tank body 1 is made of high-silicon glass, and its top cover is made of metal.

[0031] Furthermore, the rotatable spiral guide tube 3 includes a spiral baffle 31 and a guide tube 32. The guide tube 32 is fixed in the center inside the tank body 1, and the upper and lower ends of the spiral baffle are respectively rotatably and sealingly connected to the top cover of the tank body 1 and the microbubble generator.

[0032] Furthermore, the ratio of the pitch of the spiral baffle 31 to the height of the guide tube is 1:1, the ratio of the spiral diameter to the diameter of the guide tube is 4:5, and the ratio of the diameter of the guide tube 2 to the diameter of the microbubble generator is 4:3.

[0033] Preferably, the spiral baffle 31 includes a gradient spiral baffle body. The bottom of the gradient spiral baffle body is embedded in the center of the microbubble generator via a sealed bearing, and the top is mounted on the top cover of the tank via a sealed bearing to prevent the culture medium from seeping into the bearing and causing contamination. The bottom pitch of the gradient spiral baffle body is relatively large, allowing the initial bubbles to enter smoothly and reducing resistance. As the bubbles rise, the pitch decreases, promoting bubble breakage and maintaining a small bubble size.

[0034] As a preferred embodiment, the surface equation of the gradient spiral baffle body is:

[0035] z = -0.0333x 2 -0.0333y2+0.0667xy-3.333x+0.0005y+0.0012

[0036] x∈(-45,45); y∈(-45,45);

[0037] Wherein, the origin o is the geometric center of the circular structure at the bottom of the spiral baffle, the positive x-axis is the extension direction along the tangent of the outer edge of the bottom spiral baffle, the positive y-axis is the direction of rotating 90° counterclockwise along the positive x-axis, and the positive z-axis is the direction extending upward perpendicular to the xoy plane. As an improvement, the airlift bioreactor also includes a temperature control jacket 2, which surrounds the outer circumference of the tank 1. The temperature control jacket 2 is connected to the constant temperature water tank 13 via a circulating water pump 14. The circulating water pump circulates water at a fixed temperature in the constant temperature water tank through the temperature control jacket, and then uses internal circulation within the tank for heat transfer to control the tank temperature.

[0038] As a further improvement, the airlift bioreactor also includes a temperature probe 12, which is installed inside the tank 1 through a temperature probe port provided on the top cover of the tank.

[0039] As an improvement, the airlift bioreactor also includes a dissolved oxygen electrode 10 and a pH electrode 11, which are installed inside the tank 1 through dissolved oxygen electrode ports and pH electrode ports provided on the top cover of the tank, respectively.

[0040] As an improvement, the airlift bioreactor further includes a precursor feed bottle 15, an inducer feed bottle 16, a feedback inhibitor feed bottle 17, and an acid / base feed tank 22. The top cover of the tank 1 is respectively provided with an acid / base feed port, a precursor / inducer feed port, and a feedback inhibitor feed port. The precursor feed bottle 15 is connected to the precursor / inducer feed port through a precursor feed peristaltic pump 18, the inducer feed bottle 16 is connected to the precursor / inducer feed port through an inducer feed peristaltic pump 19, the feedback inhibitor feed bottle 17 is connected to the feedback inhibitor feed port through a feedback inhibitor feed peristaltic pump 20, and the acid / base feed tank 22 is connected to the acid / base feed port through an acid / base feed peristaltic pump 21.

[0041] As an improvement, the microbubble generator 4 is connected to an air supply device, which includes a check valve 5, a flow rotor 6, an intake air filter 7, and an air pump 8. The air pump 8 filters compressed air into sterile air through the intake air filter 7. The sterile air then passes through the flow rotor 6 to regulate the gas flow rate, and then through the check valve 5 into the microbubble generator 4 to prevent liquid backflow. The air is then dispersed into microbubbles by the microbubble generator and enters the tank as a power and oxygen source. The sterile air then passes through a rotatable spiral guide tube 3 to drive the culture medium in a spiral circulation motion. Finally, the air is discharged from the exhaust port on the tank after being filtered by the exhaust air filter 9 to prevent tank contamination.

[0042] The following describes the specific implementation method of Embodiment 1 of this utility model:

[0043] The amplification and culture of *Taxus chinensis* stem cells using an airlift bioreactor as described in Embodiment 1 of this utility model includes the following steps:

[0044] After adding an appropriate amount of water to the tank, it is sterilized by steam at 121℃ for 40 minutes, then cooled and pumped out for later use. Modified B5 culture medium with an initial pH of 5.8 and 10% inoculum of *Taxus chinensis* stem cells are added through the inlet. The culture temperature in the tank is controlled at 25℃ using a temperature control jacket. Compressed air is filtered through an air filter to obtain sterile air, and the airflow is adjusted to 0.2 vvm by a flow rotor. The air then passes through a check valve to prevent backflow, and is dispersed into microbubbles by a microbubble generator before entering the tank as a power and oxygen source. The kinetic energy of the microbubble airflow drives the spiral baffle to rotate, promoting the spiral ascent of the *Taxus chinensis* stem cell culture medium in the guide tube, followed by a descent outside the guide tube to achieve circulation. The sterile air then exits through the exhaust port, which is connected to an air filter to prevent tank contamination.

[0045] On day 6 of incubation, a peristaltic pump was used to add 400 mg / L phenylalanine, 5 mM methyl jasmonate, and 2 mg / L gibberellin to the bioreactor through the precursor / inducer feed port and the feedback inhibition feed port, respectively. Induction continued for another 8 days. The pH of the system was maintained at 5-7 during the incubation process.

[0046] Comparison Case 1:

[0047] Comparative Case 1 is an airlift bioreactor, which includes a tank, a temperature control jacket, a DC flow guide tube, and a microbubble generator.

[0048] The difference between this comparative case 1 and the aforementioned embodiment 1 is that the rotatable spiral guide tube 3 is a DC guide tube, which is a straight tube and is fixed in the center of the tank, above the microbubble generator.

[0049] The following describes the specific implementation method of Comparative Example 1 of this utility model:

[0050] The amplification and culture of Taxus chinensis stem cells using the airlift bioreactor described in Comparative Case 1 includes the following steps:

[0051] The method for amplifying and culturing Taxus chinensis stem cells involves adding an appropriate amount of water to the tank, sterilizing it with steam at 121℃ for 40 minutes, and then pumping out the water after cooling. Modified B5 medium with an initial pH of 5.8 and 10% inoculum of Taxus chinensis stem cells are added through the inlet, and the culture temperature in the tank is controlled at 25℃. Compressed air is filtered through an air filter to obtain sterile air, and the airflow is adjusted to 0.2 vvm using a flow rotor. After passing through a check valve to prevent liquid backflow, the air is dispersed into microbubbles by a microbubble generator and enters the tank as a power and oxygen source. The kinetic energy of the microbubble airflow propels the Taxus chinensis stem cell culture medium upward in the guide tube and downward outside the guide tube to achieve circulation. The sterile air is then discharged from the exhaust port after passing through the tank, and the exhaust port is connected to an air filter to prevent tank contamination.

[0052] On day 6 of incubation, 400 mg / L phenylalanine, 5 mM methyl jasmonate, and 2 mg / L gibberellin were added to the bioreactor through the precursor / inducer feed port and the feedback inhibition feed port, respectively, from the precursor feed port, inducer feed port, and feedback inhibition feed port. Induction continued for another 8 days. The pH of the system was maintained at 5-7 during the incubation process.

[0053] Comparison Case 2:

[0054] Comparative Case 2 is an airlift bioreactor, which includes a tank, a temperature control jacket, a fixed spiral guide tube, and a microbubble generator.

[0055] The difference between this comparative case 1 and the aforementioned embodiment 1 is that the rotatable spiral guide tube 3 is a fixed spiral guide tube with a spiral baffle in the middle, and is installed inside the tank. The guide tube is fixed in the center of the tank, above the microbubble generator.

[0056] The following describes the specific implementation method of Comparative Example 2 of this utility model:

[0057] The amplification and culture of Taxus chinensis stem cells using the airlift bioreactor described in Comparative Case 2 includes the following steps:

[0058] After adding an appropriate amount of water to the tank, it is sterilized by steam at 121℃ for 40 minutes, then cooled and pumped out for later use. Modified B5 culture medium with an initial pH of 5.8 and 10% inoculum of *Taxus chinensis* stem cells are added through the inlet, and the culture temperature in the tank is controlled at 25℃. Compressed air is filtered through an air filter to obtain sterile air using an air pump. The airflow is adjusted to 0.2 vvm by a flow rotor, then passes through a check valve to prevent liquid backflow, and is dispersed into microbubbles by a microbubble generator before entering the tank as a power and oxygen source. The kinetic energy of the microbubble airflow propels the *Taxus chinensis* stem cell culture medium upward in a fixed spiral guide tube, and then downward outside the guide tube to achieve circulation. The sterile air is then discharged from the outlet after passing through the tank, and the outlet is connected to an air filter to prevent tank contamination.

[0059] On day 6 of cultivation, 400 mg / L phenylalanine, 5 mM methyl jasmonate, and 2 mg / L gibberellin were added to the bioreactor, and induction continued for another 8 days. The pH of the system was maintained at 5-7 during the cultivation process.

[0060] Table 1 Comparison of stem cell culture data from Taxus chinensis var. mongolica.

[0061] Case Cell biomass (DCW g / L) Paclitaxel content (mg / L) Comparison Case 1 9.04±0.62 3.24±0.15 Comparison Case 2 12.53±0.75 4.64±0.18 Implementation Case 1 14.25±0.51 5.15±0.11

[0062] As shown in Table 1, compared with Comparative Case 1 (DC flow guide tube) and Comparative Case 2 (fixed spiral flow guide tube), the dry weight of *Taxus chinensis* stem cells in Example 1 of this invention increased by 1.58 times and 1.14 times respectively after 14 days of culture, and the paclitaxel yield increased by 1.59 times and 1.19 times respectively. Compared with traditional reactors, this invention has three major advantages: low energy consumption, high mass transfer efficiency, and adaptability to large-scale culture, providing innovative equipment support for the industrial production of plant stem cells.

[0063] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and design concept of the present invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. An airlift bioreactor for scale-up culture of Taxus chinensis stem cells, characterized in that, The device includes a tank, a rotatable spiral guide tube, and a microbubble generator. The microbubble generator is located at the bottom center of the tank, and the rotatable spiral guide tube is located inside the tank and fixed above the microbubble generator. The top cover of the tank is provided with an inlet, an outlet, and an exhaust port. The culture medium enters the tank through the inlet, and sterile air is dispersed into microbubbles by the microbubble generator and enters the tank. The kinetic energy of the microbubble airflow promotes the culture medium to spiral upward inside the rotatable spiral guide tube and spiral downward outside the rotatable spiral guide tube to achieve circulation.

2. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 1, characterized in that, The rotatable spiral guide tube includes a spiral baffle and a guide tube. The guide tube is fixed in the center inside the tank. The upper and lower ends of the spiral baffle are rotatably and sealingly connected to the top cover of the tank and the microbubble generator, respectively.

3. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 2, characterized in that, The ratio of the pitch of the spiral baffle to the height of the guide tube is 1:1, the ratio of the spiral diameter to the diameter of the guide tube is 4:5, and the ratio of the diameter of the guide tube to the diameter of the microbubble generator is 4:

3.

4. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 2, characterized in that, The spiral baffle includes a gradient spiral baffle body, the bottom of which is embedded in the center of the microbubble generator through a sealed bearing, and the top of which is mounted on the top cover of the tank through a sealed bearing.

5. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 1, characterized in that, It also includes a temperature control jacket, which surrounds the outside of the tank.

6. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 5, characterized in that, The temperature control jacket is connected to the constant temperature water tank via a circulating water pump.

7. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 1, characterized in that, It also includes a temperature probe, a dissolved oxygen electrode, and a pH electrode; the temperature probe, dissolved oxygen electrode, and pH electrode are installed inside the tank through the temperature probe port, dissolved oxygen electrode port, and pH electrode port set on the top cover of the tank, respectively.

8. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 1, characterized in that, It also includes a precursor feed bottle, an inducer feed bottle, a feedback inhibitor feed bottle, and an acid / base feed tank. The top cover of the tank is respectively equipped with an acid / base feed port, a precursor / inducer feed port, and a feedback inhibitor feed port. The precursor feed bottle is connected to the precursor / inducer feed port through a precursor feed peristaltic pump, the inducer feed bottle is connected to the precursor / inducer feed port through an inducer feed peristaltic pump, the feedback inhibitor feed bottle is connected to the feedback inhibitor feed port through a feedback inhibitor feed peristaltic pump, and the acid / base feed tank is connected to the acid / base feed port through an acid / base feed peristaltic pump.

9. The airlift bioreactor for scale-up culture of Taxus chinensis stem cells as described in claim 1, characterized in that, The microbubble generator is connected to an air supply device, which includes a check valve, a flow rotor, an intake air filter, and an air pump. The air pump filters compressed air through the intake air filter to produce sterile air. The sterile air then passes through the flow rotor to regulate the gas flow rate, and finally enters the microbubble generator through the check valve.

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