Plantform Bioreactor

TECHNICAL DESCRIPTION

Conventional micropropagation is expensive and labour intensive. The production systems in use for the last 20 years are still in use by most laboratories. Until now few laboratories have been truly profitable. However with development of a new biological pathway using liquid culture systems in bioreactors it is now possible that micropropagation become profitable. A small in vitro culture container, named RITA, based on this system was developed by a French company and patented in France 1995. This container was mainly developed for somatic embryogenesis and is not suitable for large scale in vitro plant propagation.

We have developed a commercially attractive bioreactor for large scale in vitro plant production. The bioreactor has been designed in a way that is easy to handle, it has low weight, it is transparent, it is autoclavable, the gas exchange including oxygen and carbon dioxide can be controlled using air pumps and a timer. The units can be places above each other saving place in the climate chamber. Fig 1 shows the construction and the details of the bioreactor.

FIG: 1.1

Bioreactor and its different parts numbered from A-F. 1:2 Outer container with 3 inlets/outlets for gas exchange, E shows the middle filter connected to a plastic tube on the inner chamber, 1:3 filters, plastic tubes, nuts, clamps and silicon rings to be connected to the 3 inlets/outlets on the outer container, 1:4A inner chamber with 3 grooves on the long side and connection to the middle filter, 1:5B basket with 3 rows of small holes, 1:6C frame with 4 legs, 1:7 lid with 4 flaps and an inner silicon ring.

Description

The bioreactor consists of a body made of polycarbonate which is transparent and can stand heating 120 C. The size is 180x150x150 cm. Gas exchange is controlled through three inlets/outlets anchored to the side by nuts and clamps through holes in the body and sealed with heat resistant silicon o-rings. The gases are passed through sterile filters connected by silicon tubes to the inlets. The middle filter is connected to a plastic tube on the inner chamber and goes through the basket (fig 2 grey)
An inner chamber with 3 grooves on the long side and 2 grooves on short side is placed at the bottom of the bioreactor. It is designed in a way to allow nutrients to rise efficiently when a pressure is applied (fig 2 purple).
A basket containing the plant material is placed above the inner chamber. The small holes in the basket is placed in three rows which allow the nutrients to flow efficiently through the basket (fig 2 grey)
A frame with four legs is placed at the bottom of the basket to press down the basket when a pressure is applied. (fig 2 red)
The body is closed by a lid which is air tight using silicon seal inside a groove of the lid. The lid is a snap-on lid easy to put on and remove.
Number 3, 4 and 5 are made of white polypropylene which can be autoclaved at 120 C.

Description

After sterilizing all parts the bioreactor is ready to use. The body is filled with generally 500ml of nutrient solution under the inner chamber. Then sterile plant material is placed on the basket. The frame with the four legs are placed on the basket and the lid is secured at the top of the body. The filters are connected to a dual timer and a compressor with solenoid valves to regulate gas exchange or air pumps. When a pressure is applied to the middle filter the nutrient solution is forced upwards and cover the plant material. When pressure is relieved the nutrients will drain back through the holes in the basket. No liquid will be left on the basket which is important to avoid hyperhydricity. The immersion time and frequency is set by a timer. The other filters are used for ventilation and CO2 supply. The duration and frequency of ventilation is set by a timer. Since the body has feet at the bottom several units can be placed above each other and connected to a single control unit.

The Advantage of Plantform Bioreactor

The bioreactor will reduce production costs and improve plant quality.
Less rigid temperature control.
Labour required will be reduced as handling and cutting will be reduced.
The plant are produced larger fully functional leaves and roots which reduce the need for weaning.
Plants produced are larger in size which accelerate their growth in the substrate.

Figure 4 shows several bioreactors connected to the control unit and simple air pumps.

KEY POINTS

HANDLING OF PLANTFORM BIOREACTOR

Do not use strong detergents during washing of the bioreactors (The plastic can get white spots).
We recommend 115-120° C and 20 min. for autoclaving the bioreactors (temperature above 120o C can damage the lids). We recommend sterilizing the filters separately because steam can block the membrane. The filters can be wrapped in foil, paper or put in a closed glass jar during autoclaving. The tubes on the bioreactor should be wrapped with aluminium foil during autoclaving.
It is more efficient to autoclave the medium in the bioreactor. However if you choose to autoclave the bioreactors empty then you can leave the filters on the bioreactor during autoclaving but wrap them with aluminium foil.
Close only two sides of the lid during autoclaving.
If you have clean cultures and are very good in handling you do not need the autoclave the bioreactor every time you change medium.
It is possible to use a microwave for sterilizing the bioreactor and the medium. The microwave settings depend on type and age of the microwave but as a rule of thumb you can use time for boiling plus half of that time. The tubes on the bioreactors have to be wrapped in kitchen cling film. Filters must be autoclaved.
The lid has a limited length of live. If the lid does not close properly then you can get infections. The lid must then be replaced or the bioreactor wrapped with cling film around the lid. Press lid firmly in the groove and then use the flaps to close the lid.
It is also good to use the cling film if you have problems with the culture room.

Plantform bioreactor developed for in vitro plant culture and large scale micropropagation.

ajsayegh@tcpropagation.com +353873113811

Plantform BIOREACTOR

TECHNICAL DESCRIPTION

FIG: 1.1

Description

Description

The Advantage of Plantform Bioreactor

Figure 4 shows several bioreactors connected to the control unit and simple air pumps.

KEY POINTS

HANDLING OF PLANTFORM BIOREACTOR

TECHNICAL

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