Culture of Isolated Single Cells from <Emphasis Type="Italic">Taxus</Emphasis> Suspensions for the Propagation of Superior Cell Populations |
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Authors: | Michael?C?Naill Email author" target="_blank">Susan?C?RobertsEmail author |
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Institution: | (1) Department of Chemical Engineering, 159 Goessmann Laboratory, University of Massachusetts-Amherst, 686 North Pleasant Street, Amherst, MA 01003, USA |
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Abstract: | Single cells isolated from aggregated Taxus cuspidata cultures via enzymatic digestion were grown in suspension culture. High seeding density (4×105 cells/ml) and the addition of cell-free conditioned medium were essential for growth. Doubling the concentration of the nutrients
ascorbic acid (150 g/l), glutamine (6.25 mm), and citric acid (150 g/l)] had no effect on single cell growth or viability. A specific growth rate of 0.11 days−1 was achieved, which is similar to the observed growth rate of aggregated Taxus suspensions. The biocide, Plant Preservative Mixture, added at 0.2% (v/v) to all single cell cultures to prevent microbial
contamination, had no significant effect on growth or viability. Following cell sorting, single cell cultures can be used
to establish new cell lines for biotechnology applications or provide cells for further study. |
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Keywords: | cell culture flow cytometry paclitaxel protoplast Taxol Taxus |
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