GROWTH AND BIOARTIFICIAL CHARACTERISTICS OF TAXUS BACCATA SUSPENSION CULTURE (CULTIVATION IN FLASKS AND BIOREACTOR)

Любовь Васильевна ОРЛОВА; Елена Борисовна ГЛОБА; Елена Викторовна ДЕМИДОВА; Мария Владимировна ТИТОВА; Алексей Евгеньевич СОЛОВЧЕНКО; Роман Владимирович СЕРГЕЕВ; Александр Михайлович НОСОВ; Наталья Даниловна ЧЕРНЯК

GROWTH AND BIOARTIFICIAL CHARACTERISTICS OF TAXUS BACCATA SUSPENSION CULTURE (CULTIVATION IN FLASKS AND BIOREACTOR)

Любовь Васильевна ОРЛОВА, Елена Борисовна ГЛОБА, Елена Викторовна ДЕМИДОВА, Мария Владимировна ТИТОВА, Алексей Евгеньевич СОЛОВЧЕНКО, Роман Владимирович СЕРГЕЕВ, Александр Михайлович НОСОВ, Наталья Даниловна ЧЕРНЯК

Abstract

One of the most promising trends of modern phytobiotechnology is the usage of cultures of tissues and cells of higher plants for production of biologically active substances. The plants of Taxus kind are of considerable interest for the researchers as they are a source of anticancer agents. The goal of the research was to carry out a comparative analysis of growth and biosynthetic characteristics of Taxus baccata suspension culture. A strain of suspension cell culture of Taxus baccata was used as the object of research. Initial T. baccata callus culture was obtained in the Institute of Plant Physiology of the Russian Academy of Sciences out from the segments of the female plant shoots of T. baccata. A line of suspension culture cells was obtained in the Institute of Plant Physiology of the Russian Academy of Sciences out from callus cultures of cells of T. baccata. The cycle time of subculturing was 3-4 weeks. A bubble conical bioreactor (2.0 total volume) was used for hardware cultivation. Growth index, specific growth rate, productivity of cells biomass were calculated for the studied cultures. Results. It was revealed that the viability of cells was maintained at 93-95 % at the beginning of cultivation cycle and decreased slightly by the end of cultivation (up to 82-83 %). The maximum accumulation of biomass in dry cell weight was observed in 24-27 days (8.3 g / l). The studied line is characterized by a "step" on the 9 – 11th days, which is associated with the presence of two subpopulations of cells in culture or with peculiarities of utilization. No lag phase was found in all the three investigated parameters. Phase degradation did not occur even after the 36th day of cultivation. To assess the content of toxoids, a HPLC analysis of alcohol extracts of biomass cell culture of T. baccata, grown in flasks and bioreactor, which showed a peak corresponding to the retention time of paclitaxel standard, was conducted. The results of the chromatographic identification of the detected compounds were confirmed by the data of UV spectra of the corresponding to them chromatographic peaks. Conclusion. Phytochemical analysis results indicate that suspension culture of cells of yew contains the taxoids (paclitaxel and baccatin III). Synthesis of taxoids is remained in terms of passage to the hardware growing of this cell culture using a bubble bioreactor.

Keywords

plant cell culture in vitro; HPLC; taxol; baccatin III; Taxus baccata

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