Selective detachment of mitotic cells by hypotonic salt solution |
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Authors: | Edwin V Gaffney Edward G McElwain |
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Institution: | (1) Department of Biology, The Pennsylvania State University, 16802 University Park, Pennsylvania, 16802 |
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Abstract: | Summary A method has been developed to obtain synchronous populations from a human cell line which previously resisted the use of
the selective harvest technique. A concentration of Colcemid was determined which reversibly enriched the mitotic population
but avoided delays in cell cycle progression. Mitotic cells were then detached from monolayer cultures by brief treatment
with hypotonic salt solutions. The resulting populations of line A244 were shown to be viable and syntchronous by following
attachment efficiency and cycle time and by monitoring mitotic index and deoxyribonucleic acid synthesis. Hypotonic solutions
offer no advantage in the selection of mitotic L-929 cells, a line commonly synchronized by selective harves. However, their
use with both CV-1 and A244 cells provided large populations highly synchronized with respect to mitosis. This technique might
be applied successfully to cell types which do not demonstrate a selective advantage at division. |
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