Study of growth characteristics of callus, suspension and root crops Scutellaria baicalensis

Scutellaria baicalensis is a popular traditional plant in Chinese medicine. Widely distributed biologically active substances of Scutellaria baicalensis are flavonoids (baicalin, baicalein, vogonin and vogonoside), which are responsible for the antitumor activity. Their antitumor effect is due to the absorption of oxidative radicals, the weakening of the activity of NF-kB (nuclear factor-kB), the suppression of the expression of the COX-2 gene and the regulation of the cell cycle. In addition, baicalein, baicalin, and vogonin showed strong antioxidant activity. The root culture in vitro of the medicinal plant Scutellaria baicalensis is characterized by intensive growth. The growth index at the end of the cultivation cycle was 40. The growth curve has a standard S-shape, with pronounced growth phases. The stationary phase was observed from 5-7 weeks of cultivation. The root culture growth index was 22.

It is known that baicalein is used in Asian medicine (in China and Japan) for the treatment of ischemic diseases [6]. It has also been proven that this compound has antioxidant activity. In the work of J. Y. Jeong, a study was conducted on the presence of the protective effect of baicalein on DNA damage and apoptosis, as a result of which, scientists proved that baicalein effectively inhibited H2O2-induced cytotoxicity and DNA damage by inhibiting the accumulation of reactive oxygen species (ROS) [2].
Vogonin-5,7-dihydroxy-8-methoxyflavone, a flavonoid-like chemical compound, is a flavone that has an antitumor effect, which consists in inhibiting the growth of cancer cells, by stimulating autophagic and apoptotic cell death [7].
Another valuable flavonoid is oroxylin A, which induces apoptosis in human colon cancer cells via the mitochondrial pathway [5], which has an anti-hepatic effect. In the study of H. Jin, it was proved that oroxylin A relieved alcoholic liver damage by inhibiting the aging of hepatocytes, in addition, it was proved that this compound also has anti-inflammatory, anti-cancer, antibacterial and antiviral effects [4]. At the first stage of studying the growth index of callus cultures, the callus cell cultures were characterized by studying the weight of raw biomass.

Objects and methods of research
To determine the initial weight of the culture (the weight of the transplant), it was weighed before transplanting into a culture vessel (Petri dish) with a medium. After placing the graft on the medium, a second weighing was performed and the weight of the graft was determined as the difference between the second and first weighing. The weight of the grafts was equalized with an accuracy of ± 10 %. For callus cell cultures the weight of the graft was 0.1 g.
To determine the weight of the raw biomass of the culture during cultivation, the culture was removed from the culture vessel and weighed on an analytical balance. For the most stably growing suspension lines, the growth characteristics were comprehensively studied for all the parameters studied (dry weight, cell viability).
To determine the content of raw and dry biomass in a liter of medium, a fixed volume of suspension (not less than 15 ml, in three replications) was filtered through a paper filter using a Buchner funnel under vacuum. The biomass was dried to a constant weight in a current of air at a temperature of 30 ° C.
The viability of cell cultures was determined using the in vivo dye phenosafranin (0.1% solution), or 0.025 % Evans blue, by counting live (unpainted) and dead (colored) cultured units under a microscope.
Based on the results obtained, the growth index (I), the specific growth rate in the exponential phase (µ), the economic coefficient (Y), and the doubling time (τ) were calculated using the following formulas 2.3.2 and 2.3.3: Where: Xmax -the maximum value of one of the growth criteria (in this work -the content of dry biomass in a liter of medium); X0 -the initial value of one of the growth criteria.
where X2 is the content of dry biomass in a liter of medium at time t2; X1 -the content of dry biomass in a liter of medium at time t1.
At the final stage, examinations were made to study the growth characteristics of root crops in vitro. To characterize the growth of root crops in vitro, the growth index was used.
Statistical data processing was performed using the Microsoft ® Excel computer program. The text shows the arithmetic averages of the parameters. The bars in the diagram correspond to the maximum values of confidence intervals at a 95 % probability level according to the Student's tcriterion. All experiments were performed at least three times.

Results and discussion
For the obtained selected steadily growing callus cell cultures, a preliminary analysis of the growth of the cultures was performed and the growth index was calculated. The results of the study of the growth index are presented in Table 1. The main growth characteristics of suspension cultures of medicinal plants are calculated, they are presented in Table 2.  At the final stage of the task of studying the growth characteristics, root cultures of Scutellaria baicalensis were studied in vitro.
To characterize the growth of root crops in vitro, the growth index was used. The results of the research are shown in Figure 2. Biomass growth, г Duration of cultivation, day