Image analysis assessments of perfluorocarbon- and surfactant-enhanced protoplast division |
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Authors: | P Anthony M R Davey J B Power C Washington K C Lowe |
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Institution: | (1) Department of Life Science, University of Nottingham, University Park, NG7 2RD Nottingham, UK;(2) Department of Pharmaceutical Sciences, University of Nottingham, University Park, NG7 2RD Nottingham, UK |
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Abstract: | Image analysis has been used to assess the growth of cell suspension-derived protoplasts of Petunia hybrida cv. Comanche at an interface between aqueous culture medium (KM8P), supplemented with 0.01% (w/v) Pluronic F-68, and oxygenated (10 mbar; 10 min) perfluorodecalin. Protoplasts synthesised a new cell wall and entered normal mitotic division which was sustainable to the cell colony/callus stage. This process was accentuated by the collective and additive effects of oxygen, perfluorodecalin and surfactant media supplements. The mean area (mm3) of protoplast-derived cell colonies after 68 days of growth was increased 35 fold over control (media alone) in the presence of these combined treatments. The new cultural regime, leading to improved cell throughput from protoplasts, is discussed primarily in relation to the role of perfluorodecalin as a gas carrier and possible effects of Pluronic F-68 in stimulating cellular uptake of nutrients and/or growth regulators. Image analysis provides a novel and accurate approach to quantifying cell growth responses.Abbreviations dpi
dots per inch
- FPE
final plating efficiency
- IPE
initial plating efficiency
- KM
Kao & Michayluk (1975)
- PFC
Perfluorocarbon
- UM
Uchimiya & Murashige (1974) |
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Keywords: | cell culture image analysis Petunia hybrida protoplast perfluorocarbon surfactant |
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