Biotechnology Letters

, Volume 27, Issue 21, pp 1725–1730

Culture of Isolated Single Cells from Taxus Suspensions for the Propagation of Superior Cell Populations

Article

DOI: 10.1007/s10529-005-2738-1

Cite this article as:
Naill, M.C. & Roberts, S.C. Biotechnol Lett (2005) 27: 1725. doi:10.1007/s10529-005-2738-1

Abstract

Single cells isolated from aggregated Taxuscuspidata cultures via enzymatic digestion were grown in suspension culture. High seeding density (4×10cells/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.

Key words

cell cultureflow cytometrypaclitaxelprotoplastTaxolTaxus

Copyright information

© Springer 2005

Authors and Affiliations

  1. 1.Department of Chemical Engineering, 159 Goessmann LaboratoryUniversity of Massachusetts-AmherstAmherstUSA