Pneumatic self-circulation animal cell culture bioreactor and application method thereof

Abstract

The invention discloses a pneumatic self-circulation animal cell culture bioreactor and an application method thereof. The bioreactor is characterized by comprising a large tank, a small tank located in the large tank, a magnetic stirring system located at the bottom of the large tank, a heating system and a controlling system, wherein a sealing cover is arranged at the upper end of the large tank, jackets connected with the heating system are arranged on two sides of the large tank, and a highest liquid level sensor of the large tank is arranged at the upper part in the large tank; and a sealing head is arranged at the upper end of the small tank, an opening is formed in the lower end, the sealing head is fixedly connected in a sealing manner with the sealing cover through a breathing pipe, the upper end of the breathing pipe penetrates out of the sealing cover and is connected with a vacuum air pump and an air pressure pump respectively, the lower end penetrates out of the sealing head and enters the small tank, a cavity used for accommodating a cell carrier is defined by two up-and-down horizontal stainless steel wire mesh boards in the middle of the small tank and the inner wall of the small tank, an upper liquid level sensor of the small tank is arranged above the cavity, and a lower liquid level sensor of the small tank is arranged below the cavity. The pneumatic self-circulation animal cell culture bioreactor has the advantages of good air tightness, simplicity in operation, low contamination rate, good cell quality and high yield.

Description

technical field [0001] The invention relates to an animal cell culture device, in particular to a pneumatic self-circulation animal cell culture bioreactor and its application method. Background technique [0002] Bioreactor refers to equipment that uses living cells or enzymes as biocatalysts to provide a suitable environment for cell proliferation or biochemical reactions. It is the key equipment in the process of biological reactions. The selection of the structure, operation mode and operating conditions of the bioreactor is closely related to the quality, yield (conversion rate) and energy consumption of biochemical products. With the continuous development of biotechnology, large-scale animal cell bioreactor culture technology is gradually replacing glass bottle static culture and rotary culture technology, because it can provide the best living environment for the cultured objects in time, so that the most efficient It should be said that this is a qualitative leap i...

AI Technical Summary

Problems solved by technology

With the continuous development of biotechnology, large-scale animal cell bioreactor culture technology is gradually replacing glass bottle static culture and rotary culture technology, because it can provide the best living environment for the cultured objects in time, so that the most efficient It should be said that this is a qualitative leap in the history of animal cell culture and a major breakthrough in biopharmaceutical technology. It not only simplifies the process, but more importantly, it can produce high-quality products. The current market There are many kinds of bioreactors on the market, including domestic and imported ones, each has its own advantages and disadvantages, and some even have unavoidable flaws, which limit the maximum use of its advantages

[0003] The name of the Chinese invention patent is animal cell culture bioreactor with external circulation (application number CN201010242025.4), which discloses a bioreactor including a tank, an external circulation pipeline, a heater, a peristaltic pump and a control system. The tank Including large tanks and small tanks, the small tank sleeve is placed inside the large tank, and is respectively fixed by pipes D, A, G and the large tank ferrule. The pipeline A and pipeline B connected with the external circulation pipeline of the tank, the pipeline E and the pipeline D connected with the external circulation pipeline of the tank through a tee; the external circulation pipeline of the tank passes through the peristaltic pump; the pipeline D is in its The lowest position is connected to the sewage pipe, which has the characteristics of complete sterilization, simple operation, and low cell culture contamination rate, but has the following disadvantages: 1) Design principle: the liquid in the large tank flows through the small tank cells through an external circulating pump and then returns to the cells. For large tanks, since the liquid always flows preferentially from the place with the least resistance, and the "channeling phenomenon" in the gas-liquid system, it will form a local dead corner of cell growth, cause local cell apoptosis, and affect the product yield; 2 ) Circulation power: use an external circulation pump to circulate, which has a large shear force on cells or products, which is not conducive to further application; 3) Stirring method: the tangential impulse when the liquid is transported back to the large tank through the external circulation pump drives the large tank The liquid is rotated to mix, which will cause uneven mixing of the upper and lower sides of the large tank and the liquid on both sides; 4) Ventilation mode: direct deep ventilation, many large bubbles, shear force on the target, large ventilation volume, and often Dissolved oxygen cannot be guaranteed, and in anoxic state, lactic acid increases, which will consume a large amount of sodium bicarbonate, and the pH of the liquid is often uncontrollable, so it can only be controlled by adding a large amount of sodium bicarbonate liquid, and the osmotic pressure of the liquid will decrease. 5) Structural design: There are many vertical pipelines, and the overall structural design is complex, and an external circulation pump is used for circulation. In this circulation mode, each pipeline will generate fatigue stress under long-term working conditions , which will affect the circulation per unit time

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