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A freeze-preservation of synchronously dividing cultured cells of Acer pseudoplatanus L
Authors:L A Withers
Institution:Botanical Laboratories, University of Leicester, Leicester, LEI 7RH, England
Abstract:A suspension culture of sycamore (Acer pseudoplatanus L.) was synchronised in division by release from nitrogen starvation. Cell samples, taken during the lag phase and synchronous growth, were examined cytologically and subjected to a freeze-preservation protocol. A high positive correlation was found between mitotic index (percentage of cells showing mitotic figures) and cell survival, as measured by fluorescein diacetate staining and reduction of 2,3,5-triphenyl tetrazolium chloride. Specific staining of lethally damaged cells and subsequent examination of the surviving cells demonstrated that the latter had a lower mitotic index and more consistent, low value of nuclear DNA than the total population. This indicated that it is cells, newly entered into G1 phase of the cell cycle, which are particularly resistant to the stresses imposed by the freeze preservation protocol.
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