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The cell culture medium — a functional extracellular compartment of suspension-cultured cells

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Abstract

The spent medium of suspension-cultured cells of Lupinus polyphyllus was analyzed by capillary GC and GC-MS and shown to contain ethanol (up to 160 mmol l-1), organic acids (lactate, benzoate, succinate, fumarate, malate), amino acids (main components: alanine, glycine, serine, aspartate, ornithine, glutamate), and quinolizidine alkaloids (lupanine and an uncharacterized malonylderivative). In addition, cells obviously secrete polysaccharides and enzymes (acid phosphatase, phosphodiesterase, DNAse, esterase, α-mannosidase, α-galactosidase, β-glucosidase, lipase, protease and peroxidase) into the medium. Typically these enzymes are localized in the vacuole of intact cells. Cytosolic enzymes, such as glutamate dehydrogenase and malate dehydrogenase were retained by the cells. Peroxidase is overexpressed in suspension-cultured lupin cells but only one basic isoenzyme is secreted, whereas the others are retained in the vacuole. In lupin leaves this isoenzyme is sequestered in the vacuole, implying that secretion is selective and needs a change in the sorting signals of the peroxidase protein. The cell culture medium shares many features of the vacuole. We assume therefore that the medium functions as a lytic compartment. In addition it provides a sink-source system for nutrients and metabolites.

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Abbreviations

ADH:

alcohol dehydrogenase

FW:

fresh weight

GC:

gas chromatography

GC-MS:

gas chromatography — mass spectrometry

POD:

peroxidase

QA:

quinolizidine alkaloids

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  1. Institut für Pharmazeutische Biologie, Universität Heidelberg, Im Neuenheimer Feld 364, D-69120, Heidelberg, Germany

    Michael Wink

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Wink, M. The cell culture medium — a functional extracellular compartment of suspension-cultured cells. Plant Cell Tiss Organ Cult 38, 307–319 (1994). https://doi.org/10.1007/BF00033891

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