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Cancer Cell death Cell cycle Cytoskeleton Exo/endocytosis Differentiation Division Organelles Signalling Stem cells Trafficking
Cell Biology International (2006) 30, 262–269 (Printed in Great Britain)
Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata
A. Yari Khosroushahia, M. Valizadehb, A. Ghasempourc, M. Khosrowshahlid, H. Naghdibadie, M.R. Dadpourb and Y. Omidia*
aResearch Centre for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran
bFaculty of Agriculture, Tabriz University, Tabriz, Iran
cResearch Centre for Medicinal Plants, Shaheed Beheshti University, Tehran, Iran
dFaculty of Natural Sciences, Tabriz University, Tabriz, Iran
eInstitute of Medicinal Plants, Tarbiat Modarres University, Tehran, Iran


Abstract

To date enormous attempts have been devoted to improve Taxol production exploiting various methodologies from bioprocess engineering to biotechnological and synthetic approaches. We have developed a 2-stage suspension cell culture of Taxus baccata L. using modified B5 medium in order to improve cell growth as well as productivity. After callus induction and cell line selection, B5 medium was supplemented with vanadyl sulfate (0.1mg/l), silver nitrate (0.3mg/l) and cobalt chloride (0.25mg/l) at the first day of stage I culture to maximize cell growth. This medium was further supplemented with sucrose (1%) and ammonium citrate (50mg/l) on day 10 and sucrose (1%) and phenylalanine (0.1mM) on day 20 (i.e., biomass growth medium). At stage II (day 25), two different concentrations of several elicitors such as methyl jasmonate (10 or 20mg/l), salicylic acid (50 or 100mg/l) and fungal elicitor (25 or 50mg/l) were added to the biomass growth medium with the aim of improving cellular productivity.

For morphological analysis, microscopic inspection was carried out during cultivation. Cell-associated and extracellular amount of Taxol were detected and measured using HPLC methodology. At stage I, overall Taxol amount of biomass growth medium was 13.75mg/l (i.e., 5.6-fold higher than that of untreated B5 control). At stage II, treated cells with methyl jasmonate (10mg/l), salicylic acid (100mg/l) and fungal elicitor (25mg/l) produced the highest amount of Taxol (39.5mg/l), which is 16-fold higher than that of untreated B5 control (2.45mg/l). Microscopic analyses of Taxus cells in suspension cultures showed various positional auto-fluorescence showing direct correlation with Taxol production. Our studies revealed that intervallic supplementation of B5 medium with combination of biomass growth factors at stage I and mixture of elicitors at stage II could significantly increase Taxol production. Thus, we suggest that the exploitation of this methodology may improve the production of Taxol since demands for Taxol pharmaceuticals are increasingly growing and resource paucities have limited its direct harvesting from Taxus trees.


Key words: Bioprocess engineering, Plant cell, Suspension cell culture, Taxol, Taxus baccata L..

*Corresponding author.


Received 11 June 2005/30 October 2005; accepted 5 November 2005

doi:10.1016/j.cellbi.2005.11.004


ISSN Print: 1065-6995
ISSN Electronic: 1095-8355
Published by Portland Press Limited on behalf of the International Federation for Cell Biology (IFCB)