This medium was originally developed for the culture of Vicia hajastana protoplasts. The culture of low cell densities (1-2 cells/ml) is enhanced when the medium is supplemented with organic acids, sugar alcohols, sugars, plant growth regulators and amino acids. Cell growth and division could also be enhanced by raising the concentrations of calcium chloride from 1 to 5 mM.
References
Binding, H. and Nehls, R. (1978) Regeneration of Isolated Protoplasts of Vicia faba L. Z. Pflanzenpbysiol. 88, 327-332. Grzebelus E, M Szklarczyk & R Baranski (2012) An improved protocol for plant regeneration from leafand hypocotyl-derived protoplasts of carrot.
Plant Cell Tiss Organ Cult 109:101-109. Kao, KN and MR Michayluk (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126:105-110.