Abstract
Five separate cell lines, three of Taxus canadensis Marsh. and two of Taxus cuspidata Sieb. et Zucc., were used to test the effect of carbohydrates and plant growth regulators on the growth of cells and production of paclitaxel in culture. There was no significant correlation between growth of cells and paclitaxel production. While no single medium was developed that was optimal for all cell lines, it was possible to develop a medium for each species that represented a superior combination of growth and paclitaxel production. A combination of NAA and thidiazuron produced the best combination of growth and paclitaxel production in cell lines of T. canadensis, while IAA and BA produced the best results in cell lines of T. cuspidata. A mixture of sucrose and fructose gave the best combination of growth and paclitaxel production. The addition of carbohydrates midway through the growth cycle increased the rate at which paclitaxel accumulated in the culture medium. The highest paclitaxel concentration obtained was 14.78±0.86 mg 1−1 (n=3).
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Abbreviations
- 2,4-D :
-
2,4-dichlorophenoxyacetic acid
- 2ip:
-
6-(γ,γ-dimethylamino)-purine
- BA:
-
6-benzyladenine
- IAA:
-
indole-3-acetic acid
- IBA:
-
indole-3-butyric acid
- kinetin:
-
6-furfurylaminopurine
- NAA:
-
α-napthaleneacetic acid
- picloram:
-
4-amino-3,5,6-trichloropicolinic acid
- thidiazuron:
-
1-phenyl-3 (1,2,3-thiadiazol-5-yl)urea
References
Christen AA, Bland J & Gibson DM (1989) Cell culture as a means to produce taxol. Proc. Am. Assoc. Cancer Res. 30: 566
Christen AA, Gibson DM & Bland J (1991) Production of taxol or taxol-like compounds in cell culture. US Patent 5,019, 504
Cragg GM & Snader KM (1991) Taxol: the supply issue. Cancer Cells 3: 233
Denis J-N, Greene AI, Guenard D, Gueritte-Voegelein F, Mangatal L & Potier P (1988) A highly efficient, practical approach to natural taxol. J. Amer. Chem. Soc. 110: 5917
Fett-Neto AG, DiCosmo F, Reynolds WF & Sakata K (1992) Cell culture of Taxus as a source of the neoplastic drug taxol and related taxanes. Biotechnology, 10: 1572–1575
Fett-Neto AG, Melanson SJ, Sakata K & DiCosmo F (1993) Improved growth and taxol yield in developing calli of Taxus cuspidata by medium composition. Biotechnology 11: 731–734
Fett-Neto AG, Melanson SJ, Nicholson SA, Pennington JJ & Dicosmo F (1994) Improved taxol yield by aromatic carboxylic acid and amino acid feeding to cell cultures of Taxus cuspidata. Biotechnology and Bioengineering 44 (8): 966–971
Gamborg OL, Miller A & Ojima K (1968) Nutrient requirements of suspension culture of soybean root cells. Exp. Cell Res. 50: 148–151
Gibson DM, Ketchum REB, Vance NC & Christen AA (1993) Initiation and growth of cell lines of Taxus brevifolia (Pacific Yew). Plant Cell Rep. 12: 479–482
Holton RA, Liu JH, Gentile LN, & Biediger RJ (1992) Semisynthesis of taxol Proceedings of the Second National Cancer Institute Workshop on Taxol and Taxus.
Holton RA, Somoza C, Kim HB, Liang F, Biediger RJ, Boatman PD, Shindo M, Smith CC, Kim S et al. (1994) First total synthesis of taxol. 1. Functionalization of the B ring. JACS 116: 1597–1598
Kao KN & Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts plated at a very low population density in liquid medium. Planta 126: 105–110
Ketchum REB & Gibson DM (1993) Rapid isocratic reversed phase HPLC of taxanes on new columns developed specifically for taxol analysis. J. Liq. Chromatog. 16: 2519–2530
Ketchum REB, Gibson DM & Gallo LG (1995) Media optimization for maximum biomass production in cell cultures of Pacific Yew (Taxus brevifolia Nutt.). Plant. Cell. Tis. Org. Cult. 42: 185–193
Ketchum REB & Gibson DM (1995) A novel method of isolating taxanes from cell suspension cultures of yew (Taxus). J. Liq. Chromatogr. 18 (6): 1093–1111
Kim JH, Yun JH, Hwang YS, Byun SY & Kim DI (1995) Production of taxol and related taxanes in Taxus brevifolia cell cultures: Effect of sugar. Biotechnology Letters. 17(1): 101–106
Nicolaou KC, Yang Z, Liu JJ, Ueno H, Nantermet PG, Guy RK, Claiborne CF, Renaud J, Couladouros EA, Paulvannan K & Sorensen EJ (1994) Total synthesis of taxol. Nature 367: 630–634
Nitsch JP & Nitsch C (1969) Haploid plants from pollen grains. Science 163: 85–87
Pepin MF, Chavarie C & Archambault T (1991) Growth and immobilization of Tripterygium wilfordii cultured cells. Biotechnol. Bioeng. 38: 1285–1291
Sufness M (1995) Taxol: Science and Applications, CRC Press, Boca Raton, Fla.
US Forest Service (1993) Pacific Yew Draft Environmental Statement. US Department of Agriculture
Warren RS & Gould AR (1982) Salt tolerance expressed as a cellular trait in suspension cultures developed from the halophytic grass Distichlis spicata. Z. Pflanzenphysiol. 107: 347–356
Westgate PJ, Emery AH, Hasegawa PM & Heinstein PF (1991) Growth of Cephalotaxus harringtonia plant cell cultures. Appl. Microbiol. Biotechnol. 34: 798–803
Wickremesinhe ER & Arteca RN (1993) Taxus callus cultures: Initiation, growth optiminization, characterization, and taxol production. Plant Cell Tiss. Org. Cult. 35: 181–193
Wickremesinhe ERM & Arteca RN (1994) Taxus cell suspension cultures: Optimizing growth and production of taxol. Journal of Plant Physiology. 144(2): 183–188
Woo DDL, Miao SYP, Pelayo JC & Woolf AS (1994) Taxol inhibits progression of congenital polycystic kidney disease. Nature 368(6473): 750–753
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Ketchum, R.E.B., Gibson, D.M. Paclitaxel production in suspension cell cultures of Taxus . Plant Cell Tiss Organ Cult 46, 9–16 (1996). https://doi.org/10.1007/BF00039691
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DOI: https://doi.org/10.1007/BF00039691