BIOEXCEL P
Plant cell culture bioreactor!
Bioreactors have several advantages for mass cultivation of plant cells. First of all, it gives better control for scale up of cell suspension cultures under defined parameters for the production of bioactive compounds. Second, constant regulation of conditions at various stages of bioreactor operation is possible. Furthermore, handling of culture such as inoculation or harvest is easy and saves time. Also, nutrient uptake is enhanced by submerged culture conditions which Stimulate multiplication rate and higher yield of bioactive compounds. At last, number of plantlets are easily produced and can be scaled-up.
As the bioreactor is a physical/thermal system for maintenance of cells at the best culture conditions for a fast growth, several models can be considered as bioreactors, starting from simple close vessels that can be externally agitated up to complex aseptic systems controlled and regulated by appropriate software.
Plant Cell Culture?
Plants produce several different secondary metabolites, called phytochemicals mostly of them used as pharmaceuticals. In recent years biopharmaceutical/nutraceutical industry renewed increased attention in production of health‑promoting secondary metabolites using plant cell and tissue cultures.
Secondary metabolites produced in plant cells are either released into medium or accumulated in the cells. Thus the spent medium or the biomass is harvested after suitable incubation period for the extraction of bioactive compounds. Sometimes different media for growth and production are used to obtain maximum secondary metabolite production.
Cell growth and secondary metabolite production represent the main factors for selecting the suitable process mode.
The production of in‑vitro secondary metabolites can be possible through plant cell cultures. This technology represents a good model to overcome many problems linked to the conventional agriculture such as variations in the crop quality due to environmental factors
In alternative to wild collection or plant cultivation, the production of useful and valuable secondary metabolites in large bioreactors is an attractive proposal; it should contribute significantly to future attempts to preserve global biodiversity and alleviate associated ecological problems. The advantages of such processes include the controlled production according to demand and a reduced man work requirement. Plant cells have been grown in different shape bioreactors, however, there are a variety of problems to be solved before this technology can be adopted on a wide scale for the production of useful plant secondary metabolites.
Characteristic of Plant cell culture Bioreactor
Such special features of cultured plant cells necessitate to design and develop a impeller for bioreactor, which could provide proper mixing and sufficient oxygen supply throughout the bioreactor without forming dead zones and will not damage the cells during cultivation period. The hydrodynamic conditions in bioreactors are most strongly dependent on the impeller design.
Different process modes are used, such as batch culture, fed batch culture, rapid feed batch culture, two‑stage batch culture and continuous culture.
BIOEXCEL P Plant cell culture bioreactor is a system for maintenance of cells at the best culture conditions for a fast growth, several models can be considered as bioreactors, starting from simple close vessels that can be externally agitated up to complex aseptic systems controlled and regulated by appropriate software.
Bioreactors have advantages for mass cultivation of plant cells. It gives better control for scale up of cell suspension cultures under defined parameters for the production of bioactive compounds.
Plant cell culture bioreactor must control gaseous atmosphere, oxygen supply and CO2 exchange, pH, minerals, carbohydrates, growth regulators, the liquid medium rheology and cell density, for culture growth and secondary metabolite production. Our agitation systems and sterilization conditions are designed appropriately in order to control these factors.
Our impeller of Plant cell culture bioreactor is designed and developed to provide proper mixing and sufficient oxygen supply throughout the bioreactor without forming dead zones, without damaging the cells during cultivation period. The hydrodynamic conditions in bioreactors are most strongly dependent on the impeller design.
Plant cell bioreactor may use different culture methods such as batch culture, fed batch culture, rapid feed batch culture, two stage batch culture and continuous culture.
Plant cell culture bioreactor has agitation systems and pneumatically agitation system.
Mechanically stirred bioreactors have a helical ribbon impeller, magnetic stirrers, or vibrating perforated plates and a flat blade turbine impeller, which high agitation breaks incoming air into small bubbles.
Pneumatically Agitated Bioreactors have two types: bubble column and air lift. These are tall and thin in comparison with agitation bioreactors. In the bubble columns air is bubbled at the base of the column, thus medium is agitated. In air lift bioreactors, gas is sparged from the riser section to the top of the column and the medium flows downward in the down corner section.
BIOEXCEL P Bioreactor system is a lab scale plant cell culture bioreactor. This model is recommended for researcher, professor and supervisor for diversified experimental methods as well as providing intellectual communication with convenience. Usually, plant cell culture bioreactor with volume from 1L to 19L is characterized as lab scale.
BIOEXCEL M control system is basically applied for BIOEXCEL P Bioreactor. However, if user wants 4-gas mixer, ORP sensor, variable motor spped controller or any other optional functions, BIOEXCEL A controller can be applied.
The BIOEXCEL M’s control system is easy to handle, and all the monitored data and control parameters are transmitted to a PC via serial communication. The BIOEXCEL M’s 7-inch wide control panel screen, sensor port and communication port are located in the front and allows side by side placement and slim design.
BIOEXCEL M Controller PID controls pH, Temperature, Agitation speed, and DO by Cascade method. Cascade method is adopted for agitation speed or mixing gas. Fed-batch culture are available based on pH, DO. This fed-batch culture method maintains vessel inside condition based on constant level of pH, DO. Continuous culture can be also used as continuous feeding is available in the system.
BIOEXCEL M Controller supports pH, Do, temperature and foam sensor. It also includes 4 peristaltic pumps for Acid, Base, Antifoam, and feeding for fermentor.
Easy to do Batch, Fed-Batch, Continuous culture fermentation, and it enables to cultivate micro-organism by accurate adjustment of temperature, agitation speed, pH, DO, and foams (which generate during the fermentation) via microprocessor and PC. Also, these three culture modes can be used for animal cell, plant cell, and microalgae, but please note that all functions which can satisfy customer’s every single demand cannot be offered.
Sensors controlling of pH, DO, temperature and foam with 4 peristaltic pumps(acid, base, antifoam and feed), DO, pH, Temp, Foam, sensors are available.
Generally, motor for agitation is installed in top plate of vessel, and simple and durable agitation motor is used. In some case, Bottom magnetic Drive system can be applied. Various size options are available for Single vessel, Double wall vessel, Bowl vessel, Single round bottom vessel, and Small volume vessel. For cell culture such as animal or plant cell, servo motor can be used for agitation in very slow speed with accurate control of speed.
Digital PID control algorithm can control the temperature very fast, and stable temperature is maintained. pH value is calibrated in consideration of temperature, and can be controlled by feeding acid and base during fermentation. DO value is adjusted to measure accurate value according to temperature and salinity.
Description Of BIOEXCEL P Plant Cell Culture Bioreactor Vessels
SINGLE ROUND VESSEL
- Single round vessel is that vessel bottom is made of round shape, then there is no stagnant area in vessel while agitator is running.
- Heating Blanket 으로 vessel을 감싸서 온도를 조절한다. (30~40℃)
- Single round bottom vessel is used for plant cell fermentation as low speed agitation
- Single vessel is relatively cheaper than double vessel
- Low risk of glass damage from shock..
- Temperature: single vessel does not require outside temperature control equipment.
DOUBEL VESSEL
- Heat transfer is stable due to larger heat transfer surface area.
- Useful for quick cooling and decreasing temperature.
- Water Bath is required for temperature control.
- No congested area due to round shape bottom.
BOWL VESSEL
- Bottom part is stainless.
- Low risk of damage from shock.
- Production capacity : Maximum total volume 19L.
Characteristics of heat :
- Heat transfer surface area is larger than single vessel
- Temperature cooling is faster than single vessel
- It is a suitable combined form between single vessel and double vessel, and requires temperature controlling equipment (chiller or water bath)
- Bottom stainless part is double jacket which can control vessel inside temperature by heating or cooling, so cold finger is not required. For huge volume, bowl vessel is most appropriate choice
SINGLE VESSEL
- It is used for plant cell bioreactor as low speed agitation
- Single vessel is relatively cheaper than double vessel
- Low risk of glass damage from shock..
- Temperature: single vessel does not require outside temperature control equipment.
Secondary metabolites through plant cell cultivation system
There are many such secondary metabolites that can benefit the living of people as a form of pigment, medicine or perfume. However, extracting these materials directly from plants is restricted: the content is low, productivity of each part of plant is different, and the growth of plant is limited and effected by temperature, season and place. These limitations can be overcome by plant cell cultivation system. The useful, and producible secondary metabolites through plant cell cultivation system are antitumor antibiotics (vincristine, vinblastine, podophyllotoxins , maytansine, tripdio-lide, homoharringt onine, paclitaxel, ellipticin, thalicarpin, baccharin, and bouvardin), sterol as cholesterol inhibitor , heart diseases related cures ubiquinone and antibiotics, shikonin for antiulcer, and saponin in ginseng. Currently, over 300,000 species of plants are known to have secondary metabolites, and every year materials from about 1,500 plant species are extracted and separated. 300 species are evaluated to have useful physiologically active materials.
Oxygen Transfer Coefficient
The amount of O2 in bioreactor depends on the presence of O2 in the gas phase above and in the air bubbles inside the medium, as well as on the dissolved O2 in the medium. Air is released through a sparger located at the base of the bioreactor. The available oxygen for plant cells in liquid cultures, determined by oxygen transfer coefficient (kLa values), is the part that dissolves in water. The level of O2 in liquid cultures in bioreactors can be regulated by agitation or stirring and through aeration, gas flow and air bubble size. The use of a porous irrigation tube as a sparger generated fine bubbles, high kLa values, low mechanical stress and provided a high growth rate.
Photoautotrophic Cell
Plants evolve CO2 and consume O2 during respiration, while during photosynthesis CO2 is used and O2 is produced. If photoautotrophic conditions persisted in the plant cell culture, CO2 levels increased during the dark period, while they decreased during the light period. The effects of CO2 were reported for both agarized cultures and cell suspension cultures used for secondary metabolite production.
High aeration rates appear to reduce the biomass growth
In general, high aeration rates appear to reduce the biomass growth. High aeration rates were found to inhibit cell growth in cell suspensions cultured in airlift bioreactors. It is reported that high aeration rates rather than excessive oxygen inhibit growth and this reduction could be due to depletion of CO2 or to the removal of various culture volatiles including CO2. The requirement for CO2 was not related to photosynthesis but to some other metabolic pathways involved in amino acid biosynthesis.
Major factors in plant cell culture Bioreactor
There are different factors affecting the culture growth and secondary metabolite production in bioreactors: the gaseous atmosphere, oxygen supply and CO2 exchange, pH, minerals, carbohydrates, growth regulators, the liquid medium rheology and cell density. Moreover agitation systems and sterilization conditions may negatively influence the whole process. In order to manage the biomass growth in bioreactors, above mentioned factors must be controlled.
Many types of plant cell culture bioreactors have been successfully used for cultivating transformed root cultures, depending on both different aeration system and nutrient supply. Several examples of medicinal and aromatic plant cultures were here summarized for the scale up cultivation in bioreactors.
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