Biotechnology of Cultivation of Rhaponticum Carthamoides (willd.) Suspension Cells: A Prospective Source of Antitumor Substances

• Published in: Russian agricultural sciences 2022, Vol.48 (3), p.197-202
• Main Authors: Prosekov, A. Yu, Kozlova, O. V., Vesnina, A. D.
• Format: Article
• Language: English
• Subjects: Agriculture; Anticancer properties; Antitumor activity; Biological activity; Biomedical and Life Sciences; Biotechnology; Cell culture; Coffee; Composition; Crop Production; Cultivation; Cytokinins; Cytotoxicity; Growth regulators; High performance liquid chromatography; Indoleacetic acid; Kinetin; Life Sciences; Liquid chromatography; Metabolites; Methanol; Mineral composition; Nutrients; Parameters; Plant Biochemistry; Plant extracts; Plant Protection; Plant roots; Species diversity; Subculture; Tumor cells; Viability
• ISSN: 1068-3674; 1934-8037
• DOI: 10.3103/S1068367422030119

To preserve the species diversity of Rhaponticum carthamoides , a Red Book plant that is a promising source of antitumor metabolites, it is important to grow cell cultures in vitro . The aim of the work is to select the optimal nutrient medium for growing suspension cells of Rhaponticum carthamoides , varying the mineral composition of the nutrient medium and growth regulators. To obtain suspension cells in vitro , nutrient media of various mineral compositions (Murashige–Skoog, Gamborg, and Schenk–Hildebrandt) and growth regulators (auxin: 3-indoleacetic acid (IAA); cytokinin: kinetin) were used. Cultivation parameters were as follows: 26°C, 60–70% humidity, 16-h daylight period and 21-day subculture cycle. For the obtained cell cultures, the viability, growth index, and specific growth rate were assessed. The results showed that the highest values of the studied parameters were observed when growing suspension cells on a nutrient medium with a Schenck–Hildebrandt mineral composition (SH) (77–82% viability of suspension cells, 6.11‒7.14 growth index, 0.17–0.21 day –1 specific growth rate). Methanol extracts were prepared from selected and prelyophilized cell samples and analyzed by high performance liquid chromatography. The results showed that suspension cultures (the best indicators were observed in sample no. 12 grown on SH medium supplemented with IAA and kinetin), as well as the control sample (methanol extract of crushed plant root), contained sitosterol (0.26 ± 0.01 mg/g in control; 1.54 ± 0.02 mg/g in no. 12), isofraxidine (1.15 ± 0.02 mg/g in control; 3.10 ± 0.04 mg/g in no. 12), chlorogenic (44.54 ± 0.14 mg/g in control; 66.87 ± 0.02 mg/g in no. 12), and coffee (50.15 ± 0.03 mg/g in control; 54.73 ± 0.06 mg/g in no. 12) acid. The extracts of suspension cultures (100 mg/mL) exhibited antitumor properties: a cytotoxic effect on tumor cells (PANC-1, LBR2, MDAMB-231, NT-29). Cell cultures grown on SH medium supplemented with IAA and kinetin exhibited the most pronounced antitumor properties since they contained a greater amount of biologically active substances in their composition. When the extract of sample no. 12 was added, the decrease in the viability of tumor cells was achieved by 40.57–53.23%. In the course of the work, the composition of the nutrient medium was selected, which allows one to grow the biomass of suspension cultures of Rhaponticum carthamoides cells, which exhibits antitumor properties.