Articles
THE CULTURE MEDIUM
Article number
393_2
Pages
15 – 24
Language
Abstract
The selection or development of a culture medium is an essential step in any tissue culture project.
It is difficult and time consuming to optimise even a small proportion of the ingredients because of interactions in the response to different ingredients.
The selection of a medium can be simplified if the main groups of ingredients are investigated in separate experiments.
A simple protocol to determine macronutrient requirements is described, starting with a basal medium, CCB, which is then supplemented with the major nutrients.
Most cultures tolerate a wide range of micronutrient concentrations.
Whereas most organ cultures will grow on LS vitamins, protoplasts and cells, particularly at low density, may have complex requirements.
Sucrose is suitable as the sole carbon source in most media.
Optimisation of growth regulator types and concentrations is needed for each stage, but is often best handled after other components have been selected.
The choice of gelling agent often affects hyperhydricity in cultures.
It is difficult and time consuming to optimise even a small proportion of the ingredients because of interactions in the response to different ingredients.
The selection of a medium can be simplified if the main groups of ingredients are investigated in separate experiments.
A simple protocol to determine macronutrient requirements is described, starting with a basal medium, CCB, which is then supplemented with the major nutrients.
Most cultures tolerate a wide range of micronutrient concentrations.
Whereas most organ cultures will grow on LS vitamins, protoplasts and cells, particularly at low density, may have complex requirements.
Sucrose is suitable as the sole carbon source in most media.
Optimisation of growth regulator types and concentrations is needed for each stage, but is often best handled after other components have been selected.
The choice of gelling agent often affects hyperhydricity in cultures.
Authors
D. Cohen
Keywords
plant tissue culture media
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