Culture process of animal and plant cells and requirements for bioreactor
I. Introduction
Cell culture technology has been widely used in many fields such as biopharmaceutical, genetic engineering, tissue engineering, food industry and environmental science. Animal cell culture is widely used for vaccine production, antibody preparation and protein expression, while plant cell culture is mainly used for plant active ingredient extraction, genetic modification and biosynthesis. In order to optimize the culture conditions and increase the cell growth and product yield, the selection and design of bioreactor are very important. In this paper, the culture process of animal and plant cells will be described in detail and their special requirements for bioreactors will be discussed.
II, The culture process of animal cells
1. Basic steps of cell culture
- Cell preparation and inoculation
Select suitable cell lines (e.g. CHO cells, HEK293 cells, Vero cells, etc.).
After cryopreservation, the cells were cultured in a suitable medium such as DMEM, RPMI-1640 or FBS enhanced medium.
Inoculate into bioreactor or culture bottle to ensure proper inoculation density.
- Optimization of culture conditions
Temperature: Usually maintained at 36-37°C to simulate the internal environment of mammals.
pH: Controlled in 7.0-7.4, usually using CO₂ gas regulation.
Dissolved oxygen (DO) : controlled at 30-50%, maintained by agitation or gas exchange.
Nutrition supply: Provide glucose, glutamine, amino acids, vitamins and other necessary ingredients.
- Cell expansion and harvesting
Batch culture, flow-add culture or perfusion culture were used to expand the number of cells.
After the cells are cultured to optimal density, downstream products such as antibodies, proteins or viral particles can be harvested.
MIKEBIO animal cell culture bioreactor system
2. Requirements of bioreactor for animal cell culture
Animal cells are extremely sensitive to shear forces and their growth depends on a specific microenvironment, so there are the following requirements for bioreactors:
- Low shear mixing design
Magnetically stirred, air-lifted or low-speed paddles are used to reduce damage to cells from mechanical stress.
For example, Wave bioreactors can be used for suspension culture to avoid the effects of stirring paddles on cells.
- High efficiency gas exchange
Adequate oxygen should be provided, but oxidative stress caused by excessive oxygen concentration should be avoided.
Microbubble aeration or membrane aeration system is used to improve the efficiency of oxygen transfer.
- Precise environmental control
The culture environment is automatically regulated by on-line monitoring of pH, DO, temperature and CO₂ concentration.
Modern bioreactor adopts PID control system to optimize parameter adjustment.
- It is suitable for the culture of adherent cells and suspension cells
Adherent cells: Microcarriers (such as DEAE-dextran or polystyrene microcarriers) are used to provide a surface for adherent growth.
Suspension cells: Stirring or air-lift reactor can be used directly for large-scale culture.
III, The culture process of plant cells
1. Basic steps of cell culture
- Selection and preparation of cell materials
Select appropriate plant tissue (such as leaves, roots, or stem tips) for aseptic treatment.
Plant callus or cell suspension is induced and transferred into liquid medium.
- Optimization of culture conditions
Temperature: Generally controlled at 22-28°C, suitable for most plant cell growth.
pH: Usually maintained at 5.5-6.0 to facilitate cell metabolism.
Light conditions: Certain plant cells (such as algae) require light, usually using LED lighting systems.
Growth regulatory substances such as cytokinins (e.g. 6-BA) and auxin (e.g. 2,4-D) regulate cell proliferation.
- Cell expansion and accumulation of target products
The number of cells was increased by shaker culture or bioreactor culture.
The accumulation of active compounds is promoted by exogenous induction (e.g. methyl methyl jasmonate or ethylene).
Harvest cells or metabolites (e.g. alkaloids, flavonoids, terpenes).
2. Requirements of bioreactor for plant cell culture
Plant cells are more resistant to shear than animal cells, but have higher viscosity and metabolic characteristics, so there are the following requirements for bioreactors:
- Low shear mixing and ventilation system
Because plant cells are large and thick, they are susceptible to mechanical damage, so it is advisable to use airlift bioreactor or circulation stirring system.
To provide sufficient oxygen, bubble ventilation is usually used.
- Can handle high viscosity medium
Plant cell culture medium is more viscous than animal cell, so the reactor design needs to consider liquid mixing efficiency.
Efficient impellers, such as spiral impellers, are used to optimize mixing and avoid cell clumps settling.
- Precise control of growth regulators
The ratio of auxin and cytokinin is precisely controlled by an automated system to induce the production of target metabolites.
- Photobioreactor (for photosynthetic cells)
Transparent reactors or built-in LED lighting systems are used to optimize light conditions and increase the efficiency of product accumulation (e.g. chlorophyll, carotenoid production).
MIKEBIO production type fermentation system for edible fungi
Iv. Summary
Animal cell culture and plant cell culture play an important role in the biological industry, but they have significant differences in culture conditions, environmental requirements and bioreactor selection. Animal cell culture requires low shear agitation, efficient gas exchange, and precise environmental control, while plant cell culture requires high viscosity resistance, low shear agitation, and precise regulation of growth regulators. Selecting the right bioreactor for different cell types can not only improve cell survival and target product yield, but also optimize the production process and promote the development of the biotechnology industry. In the future, bioreactors combined with automation and intelligent control technology will further improve the efficiency and controllability of cell culture, and bring broader application prospects for biopharmaceuticals, agriculture and environmental protection.
On the other hand, when choosing a bioreactor fermenter, it is necessary to find a regular manufacturer with good reputation and strength, so as to ensure the quality of the product. Jiangsu Mike Biotechnology Co., Ltd. was established in 2008, is a focus on biological fermentation equipment research and development, production and sales of high-tech enterprises. Our company has a pilot fermentation platform, which is integrated with intelligence and automation, and has built a mature fermentation system, which can realize the optimization of pilot fermentation process, commissioning development of all process processes from pilot fermentation production to fermentation product separation and purification, and set up a doctor workstation and a modern laboratory.
Our company has a standardized production plant, with complete sets of equipment and tools required for installation, mainly including a variety of machine tools, automatic argon arc welding machine, automatic polishing machine, automatic cutting machine and flaw detection machine and other more than 60 sets of production and testing equipment, the production process all realize modern standard operation, to provide customers with more professional quality and technical support.