Medium plant suspension

A considerable amount of extracellular polysaccharides is produced in the process of cultivation of certain plant suspension cultures and the spent culture medium has proved to be an accessible source for their production (1-3). The interest in investigating these extracellular polysaccharides has been quite strong over the past 10-15 years, motivated by their biological activity (4,5). Plants of the Asteraceae family, as well as their cell cultures, have been established to contain polysaccharides with immunostimulating activity (1-6). The object of our research was Helianthus annuus 1805 cell culture (Asteraceae), which according to the preliminary investigation produces a considerable amount of exopolysaccharides. 

Increasing concentrations of PVP 360,000 up to 1.0 g L 1 improved antibody accumulation in hairy root culture medium however, above this concentration there was no further increase in antibody levels [19]. Addition of PVP after extracellular foreign protein levels had decreased during plant suspension culture did not result in a recovery of the protein [66]. 

A well-established plant suspension cell line. The following procedures are based on the cultivation of tobacco cells. If another cell line is chosen, the medium can be modified. 

GANTET, P IMBAULT, N THIERSAULT, M., DOIREAU, P Necessity of a functional octadecanoic pathway for indole alkaloid synthesis by Catharanthus roseus cell suspensions cultured in an auxin-starved medium. Plant Cell Physiol., 1998,39,220-225. 

Pope, D. G. Relationships between Hydroxyproline-containing Proteins Secreted into the Cell Wall and Medium by Suspension-cultured Acer pseudoplatanus Cells. Plant Physiol. 59, 894 (1977). 

The sharpness of separation of the mineral from the gangue is dependent on (1) the stability of the suspension, which is influenced by the size of the medium (2) the specific gravity of the medium (3) the cleanliness of the medium (4) the cone angle (5) the size and ratios of the internal openings in the cyclone (inlet, apex, and vortex) and (6) the pressure at which the pulp is introduced into the cyclone. A 20° cone angle is the most common. Cyclone diameter will be determined by the separation to be made as well as by the capacity required. The 0.5- and 0.6-m (20- and 24-in) cyclones are most common in coal plants, whereas multiple cones of 0.25- or 0.3-m (10- or 12-in) diameter are used in higher-gravity separations. 

Growth of the Helianthus annuus 1805 cell suspension and biosynthesis of extracellular polysaccharides. A particular characteristic of plant cell suspensions is the requirement for a high inoculation density in order to initiate growth. This is due to one of their special features in order that their growth be initiated when transferred into the new medium, they need certmn growth factors which are released and secreted into the medium by the cells themselves. Consequently, to ensure the growth of plant cell suspensions, a certain volume (in which plant cells have to be present at above certain densities) has to be used to import the necessary quantity of these substances (17). 

Besides, it is known that the culture medium acts as a common external sink like a lamella (15) or a vacuole (19), in which polysaccharides, enzymes and other metabolites are secreted during growth. Consequently, the growth of plant cell suspensions is a complex process, connected with structural and metabolite changes both in the cell wall and in the culture medium, involving a complex of hydrolytic enzymes. 

The presence of foreign protein in the medium of plant cultures does not necessarily mean that all or even most of the product can be recovered from the medium. In many expression systems where an appropriate signal sequence has been used, considerable amounts of foreign protein remain within the plant cells and/or tissues. For example, in a comparison of IgG antibody production in tobacco cell suspension and hairy root cultures, a maximum of 72% of the total antibody was found in the medium of the suspension cultures whereas only 26% was found in the medium of the hairy root cultures [17]. This result could indicate that secretion and/or transport across the cell wall was slower in the hairy roots alternatively, it could indicate poorer stability of the secreted protein in the hairy root medium. If foreign proteins are to be purified from the medium, improved secretion and extracellular product stability are desirable. 

Synonyms Natural latex—milky fluid that consists of extremely small particles of rubber obtained from plants, principally from the Hevea brasiliensis (rubber) tree, dispersed in an aqueous medium. It contains a variety of naturally occurring substances in a colloidal suspension, including about 1% proteins (e.g., a-globulins, hevein), which are the allergenic fraction. ... 

Rhizosecretion is easy to scale up and very cost effective with respect to isolation and purification. However, the bioreactor systems used for hairy root cultures differ from those used for plant cell suspensions. Traditional bioreactor systems have recently been adapted for root culture, and this technology is now being taken to commercial scales. The most traditional system is the airlift bioreactor used for microorganisms or plant cells. This system is adapted for the culturing of roots in liquid medium. Mist culture systems have also been developed. For this technology, the volume of the culture medium is reduced and the concentration of the secreted therapeutic protein is increased. If the protein to be produced is known to be quite stable, then a less expensive hydroponic culture can be designed in a manner suitable for scale-up. 

Fujita, Y., Y. Kara, C. Suga, and T. Morimoto, "Production of Shikonin Derivatives by Cell Suspension Cultures of Lithospermum erythrorhizon II. A New Medium for the Production of Shikonin Derivatives," Plant Cell Reports 1(1981 ) 61-63. 

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