Growth Culture Conditions Research Articles

Optimization of Culture Conditions for Mycelial Growth and Sporulation of Myrothecium roridum

Culture and nutrition conditions of Myrothecium roridum Tode were optimized by conducting a series of interlined experiments on a growth medium, temperature, pH, and photoperiod. In contrast, relation of culture age with virulence was measured by fungal development on young leaves of bitter gourd. The physiological response was measured on colony radial growth and spore production. Among the six test growth media, i.e., nutrient agar (NA), potato dextrose agar (PDA), Czapek-Dox agar (CDA), glucose agar (GA), malt extract agar (MEA), and bitter gourd agar (BGA), the highest radial growth (77 mm) and the highest number of spores (239 × 106 spores/ml) were observed on PDA. Incubation temperature was evaluated between a range of 15-40 °C, and the highest colony growth (87 mm) was observed at 30 °C, whereas the highest spore production (315 × 106 spores/ml) was at 35 °C. Different pH levels, i.e., 5, 5.5, 6, 6.5, 7, and 7.5, were optimized, and the highest colony growth (87 mm) and spore production (504 × 106 spores/ml) was recorded at pH 5.0. Impact of photoperiod was studied, and the highest mycelial growth (88 mm) and maximum spore production (524 × 106 spores/ml) was observed at 16/8 h alternate light and dark period. It was concluded that the optimum conditions for mycelia growth and spore production was pH 5.0-6.0 and at 30 ± 2 °C in PDA with 16/8 h alternate light and dark photoperiod.

Antibacterial activity of aquatic actinomycete,Actinoplanes digitatisDnj-2 (MN567713)

Introduction and Aim: There are centers for antibiotics in the nature to treat disease by looking for different sources, for example, actinomycetes, are approximately two-thirds of naturally occurring antibiotics, having much of medical importance, have been isolated from actinomycetes.The aim of the present study was to isolate and to identify the actinomycetes having antagonistic activity.
 
 Materials and Methods: An actinomycetes strain isolated from dam soil sample collected from Alnajl dam of Hadhramout Yemen, showed antibacterial activity against selected microbial pathogens. The nutritional requirements and cultural conditions for maximal growth to produce secondary metabolites have been optimized under shake flask conditions. The growth and production of secondary metabolites were maximal with the use of SCA medium supplemented at pH 7.0, and incubation temperature and time of 37?and 20 days respectively.
 
 Results:Based on morphological, biochemical, physiological and phylogenetic characterization, the strain was identified as Actinoplanes digitatisDnj-2 (MN567713). The ethyl acetate extract (1g/mL) obtained from the isolate showed significant antibacterial activity against Gram-negative bacteria- Escherichiacoli (21cm) and Pseudomonas aeruginosa (29cm) and Gram-positive bacteria- Bacillus subtitles(31cm) and Staphylococcus aureus (22cm) when compared with penicillin G 10 units.
 
 Conclusion:In conclusion, the isolated strain has broad spectrum of antagonistic activity against Gram-positive and Gram-negativebacteria.

Microglia Development and Maturation and Its Implications for Induction of Microglia-Like Cells from Human iPSCs.

Microglia are resident immune cells of the central nervous system and play critical roles during the development, homeostasis, and pathologies of the brain. Originated from yolk sac erythromyeloid progenitors, microglia immigrate into the embryonic brain parenchyma to undergo final postnatal differentiation and maturation driven by distinct chemokines, cytokines, and growth factors. Among them, TGFβ1 is an important regulator of microglial functions, mediating homeostasis, anti-inflammation, and triggering the expression of microglial homeostatic signature genes. Since microglia studies are mainly based on rodent cells and the isolation of homeostatic microglia from human tissue is challenging, human-induced pluripotent stem cells have been successfully differentiated into microglia-like cells recently. However, employed differentiation protocols strongly vary regarding used cytokines and growth factors, culture conditions, time span, and cell yield. Moreover, the incomplete differentiation of human microglia can hamper the similarity to primary human microglia and dramatically influence the outcome of follow-up studies with these differentiated cells. This review summarizes the current knowledge of the molecular mechanisms driving rodent microglia differentiation in vivo, further compares published differentiation protocols, and highlights the potential of TGFβ as an essential maturation factor.

Recombinant expression, purification and characterization of human soluble tumor necrosis factor receptor 2

TNFR2 is aberrantly expressed on various cancer cells and highly immunosuppressive regulatory T cells (Tregs) accumulated in tumor microenvironment. As an oncoprotein and a stimulator of the immune checkpoint Tregs that promote cancer cell survival and tumor growth, TNFR2 is considered to be a prospective target for cancer immunotherapy with the blockers developed to simultaneously inhibit abundant TNFR2+ tumor-associated Tregs and directly kill TNFR2-expressing tumors. The soluble ectodomain of TNFR2 has also been successfully applied in clinical treatment for TNF-related autoimmune diseases. Research practices on these therapeutic strategies need recombinant protein of human soluble TNFR2 (hsTNFR2); however, mass production of such biologics using eukaryotic cells is generally high-cost in culture materials and growth conditions. This study aimed to establish an efficient methodology to prepare bioactive hsTNFR2 through a prokaryotic expression system. Recombinant vector pMCSG7-hsTNFR2 was constructed and the His-tagged fusion protein expressed in E. coli was enriched in inclusion bodies. Recombinant hsTNFR2 was denatured, refolded, and then purified by affinity chromatography, tag removal, ion-exchange chromatography and gel filtration chromatography. A protein yield of 8.4 mg per liter of bacterial culture liquid with a purity of over 97% was obtained. Purified hsTNFR2 exhibited strong affinity to human TNF-α with a KD of 10.5 nM, and inhibited TNF-α-induced cytotoxicity in L929 cells with an EC50 of 0.57 μg/ml. The biological activity assessed in vitro indicated that this soluble protein can be promisingly used in drug discovery for immunotherapy of TNFR2+ cancers and treatment of autoimmune diseases featured by TNF-α overload.

A Novel Thermostable and Alkaline Protease Produced from Bacillus stearothermophilus Isolated from Olive Oil Mill Sols Suitable to Industrial Biotechnology.

This study was conducted to identify a new alkaline and thermophilic protease (Ba.St.Pr) produced from Bacillus stearothermophilus isolated from olive oil mill sols and to evaluate its culture conditions, including temperature, pH, carbon and nitrogen sources, and incubation time. The optimum culture conditions for cell growth (10 g/L) and protease production (5050 U/mL) were as follows: temperature 55 °C, pH 10, inoculation density 8 × 108 CFU/mL, and incubation time 24 h. The use of 3% yeast extract as the nitrogen sources and galactose (7.5 g/L) as the carbon sources enhanced both cell growth and protease production. Using reversed-phase analytical HPLC on C-8 column, the new protease was purified with a molecular mass of approximately 28 kDa. The N-terminal sequence of Ba.St.Pr exhibited a high level of identity of approximately 95% with those of Bacillus strains. Characterization under extreme conditions revealed a novel thermostable and alkaline protease with a half-life time of 187 min when incubated with combined Ca2+/mannitol. Ba.St.Pr demonstrated a higher stability in the presence of surfactant, solvent, and Ca2+ ions. Consequently, all the evaluated activity parameters highlighted the promising properties of this bacterium for industrial and biotechnological applications.

Morphology of filamentous thalli of Analipus japonicus (Ralfsiales), temperature and irradiance effects on their formation and growth, and erect thallus formation in the field

Free-living filamentous thalli derived from an Analipus japonicus erect thallus were established in laboratory culture. The filamentous thalli were composed of slender filaments and densely pigmented oval or sub-oval cell masses. Temperature (5–20 °C) and irradiance (25–200 μmol photons m−2 s−1) effects on formation and growth of the thalli from three explant groups (isolated filaments, small cell masses, and large cell masses) were investigated for 15 days. These explants survived over the examined range. The isolated filaments had a high ability to form the thalli, and the isolated and new filaments formed filaments and cell masses. Meanwhile, the isolated and new cell masses formed only filaments, and their ability to grow and form the thalli decreased as they grew in size. Optimum culture conditions for formation and growth of the thalli from the isolated filaments, and the two cell mass growths were 15–20 °C and 100–200 μmol photons m−2 s−1. To produce erect thalli, the filamentous thalli were cultured on seedling threads for 6 weeks at 15 °C, 60 μmol photons m−2 s−1 and 20 cm s−1 water flow velocity. Erect axis density increased with increasing coverage of the filamentous thalli, and their cell masses (primordia of erect thallus holdfast) with mean area of about 140 × 103 μm2 began to produce erect axes. After 4 months of field cultivation, they grew into full-grown erect thalli of about 20 cm long. These findings show that the free-living filamentous thalli were applicable for stable seedling production of A. japonicus to be used in aquaculture.

Molecular Profiling and Optimization Studies for Growth and PHB Production Conditions in Rhodobacter sphaeroides

In the recent climate change regime, industrial demand for renewable materials to replace petroleum-derived polymers continues to rise. Of particular interest is polyhydroxybutyrate (PHB) as a substitute for polypropylene. Accumulating evidence indicates that PHB is highly produced as a carbon storage material in various microorganisms. The effects of growth conditions on PHB production have been widely studied in chemolithotrophs, particularly in Rhodobacter. However, the results on PHB production in Rhodobacter have been somewhat inconsistent due to different strains and experimental conditions, and it is currently unclear how diverse environmental factors are linked with PHB production. Here, we report optimized growth conditions for PHB production and show that the growth conditions are closely related to reactive oxygen species (ROS) regulation. PHB accumulates in cells up to approximately 50% at the highest level under dark-aerobic conditions as opposed to light aerobic/anaerobic conditions. According to the time-course, PHB contents increased at 48 h and then gradually decreased. When observing the effect of temperature and medium composition on PHB production, 30 °C and a carbon/nitrogen ratio of 9:1 or more were found to be most effective. Among PHB biosynthetic genes, PhaA and PhaB are highly correlated with PHB production, whereas PhaC and PhaZ showed little change in overall expression levels. We found that, while the amount of hydrogen peroxide in cells under dark conditions was relatively low compared to the light conditions, peroxidase activities and expression levels of antioxidant-related genes were high. These observations suggest optimal culture conditions for growth and PHB production and the importance of ROS-scavenging signaling with regard to PHB production.

Farm or lab? Chamazulene content of Artemisia arborescens (Vill.) L. essential oil and callus volatile metabolites isolate

Plant micropropagation has been proposed as a promising solution for the improvement of medicinal plants and the commercial production of phytochemicals. Tree Wormwood (Artemisia arborescens, Asteraceae) has been identified as a promising source for chamazulene production, a phytochemical compound with anti-inflammatory activity. However, the concurrent presence of β-thujone, an acute toxic agent is a significant limitation of Tree Wormwood utilization in essential oil (EO) production. Present study is aiming to develop in vitro cultivation protocols of tree wormwood and study the effects of plant growth regulators (PGRs) and culture conditions on the callus production of volatile metabolites (VM) and its equilibrium of chamazulene and β-thujone. Three treatments all containing the auxin 1-Naphthaleneacetic acid (NAA), and each one respectively the cytokines Kinetin (K), 16-Benzylaminopurine (BN), and Thidiazuron (TD) in Murashige and Skoog (MS) basal media, were applied under constant dark (D), while K and BN were also grown under light (L). Both EO and VMs, retrieved from the naturally and laboratory grown biomass were analyzed by a Gas Chromatographer-Mass Spectrometer (GC–MS) coupled with Gas Chromatographer-Flame Ionization Detector (GC-FID). The biotechnologically produced EOs presented a significantly increased yield per area and time, while callogenesis PGRs combinations and culture conditions exhibited promising results on the induction of chamazulene biosynthesis.

Effects of culture media and physical factors on the mycelial growth of the three wild strains of Volvariella volvacea from Ecuador

Optimization of physical parameters for the growth and lipid production in Nannochloropsis gaditana (Lubian, 1982)Shyni MarKose, Ajan Chellappan, Praba Thangamani, Subilal George, Selvaraj Thangaswamy, Citarasu Thavasimuthu, Michaelbabu Mariavincent

Long-term enrichment of anaerobic propionate-oxidizing consortia: Syntrophic culture development and growth optimization

Propionate is a quantitatively important methanogenic intermediate in anaerobic digesters and only limited number of microbes can utilize it under syntrophic association with methanogens. The syntrophic propionate oxidizing bacterias are known to be slow growers due to the low energy yield. Thus, propionate get accumulated frequently in anaerobic digesters and it negatively affect digester performance. In this study, propionate degrading consortia from four different seeding sources were enriched in sequential bath mode in two phases; first adaption phase with 1 g/L of propionate concentration and later, high-strength phase with 3 g/L. From 16s rRNA gene based metagenomics analysis of the former phase, four syntrophic microbial groups, Syntrophaceae, Syntrophomonadaceae, Methanobacterium and Methanosaeta were found to be dominant with complete degradation of propionate. The substrate accelerated microbial shifts were observed at high-strength phase with significant decrease of Syntrophaceae up to 26.9 %. Using Response Surface Methodology, pH 6.8–6.9 and temperature 34.5–34.9 °C were found to be optimum growth conditions for the propionate degradation culture. Observed results could be useful to improve degradation efficiencies and obtained enriched culture can be used to recover propionate-accumulated digesters by bio-augmentation.

Enzyme free cell detachment using pH-responsive poly(amino ester) for tissue regeneration

Despite the advantages of stem cell injection to the skin wound site, this technique is limited in terms of the delivery of therapeutically functional stem cells to the wound site. Considering the abnormal microenvironment of a wound site, even a small damage or loss of cell function induced by severe trypsin treatment might lead to decrement of the therapeutic efficacy of stem cells. In this study, we have synthesized pH-responsive polymer mixed poly(amino ester) (mPAE), which can provide the appropriate cell culture condition for cell growth without cytotoxicity along with the maintenance of extracellular matrix after detachment using low pH (6.0) PBS. Human adipose derive stem cells (hADSCs) cultured on and detached from mPAE-coated tissue culture plates (TCPs) showed similar cell adhesion and angiogenic paracrine factor secretion, compared to hADSCs cultured on and detached from TCPs with trypsin. As a result, hADSCs detached with pH 6.0 PBS from mPAE-coated TCPs showed similar therapeutic angiogenesis and wound regeneration, compared to the hADSCs detached with trypsin from TCPs. In conclusion, our pH-responsive polymer, mPAE, might be utilized as an enzyme-free cell detachment system for future tissue engineering.

Cholera Toxin Production in Vibrio cholerae O1 El Tor Biotype Strains in Single-Phase Culture.

Vibrio cholerae O1 serogroup strains have been classified into classical and El Tor biotypes. Cholera, a life-threatening diarrheal disease, can be caused by either biotype through the cholera toxin (CT) that they produce. To increase our knowledge of the pathogenicity of bacteria, we must understand the toxigenicity of bacteria. CT production by classical biotype strains in simple single-phase cell cultures has been established; however, special culture media and growth conditions that are not appropriate for mass production of CT are required to facilitate CT production in El Tor biotype strains. In this report, we produced CT in El Tor biotype strains using simple media and single-phase culture conditions. A single point mutation in ToxT, a transcriptional activator of toxin co-regulated pilus (TCP) and CT, enabled the El Tor biotype strains to produce CT in similar quantities as classical biotype strains in single-phase laboratory culture conditions. CT production capacity varied between El Tor biotype strains. Wave 2 and 3 atypical El Tor strains tended to produce more CT than prototype Wave 1 strains. Wave 2 and 3 strains lack neutral fermentation; however, the capacity for neutral fermentation was not associated with significant differences in CT production by El Tor biotype strains. The Wave 3 strain that caused the 2010 cholera outbreak in Haiti produced CT only when neutral fermentation was abolished. The disparity in CT production between the seventh cholera pandemic strains highlight the differences in virulence between strains and the cause of population changes in V. cholerae.

DETERMINATION OF OPTIMUM CULTURE CONDITIONS FOR MYCELIAL GROWTH OF Macrolepiota procera MUSHROOM

Macrolepiota procera, commonly called the Parasol Mushroom, is a delicious mushroom collected from the nature and commonly consumed by the public in many regions of Turkey. This study was conducted to determine the optimum culture conditions (pH, temperature, carbon and nitrogen sources) for mycelial growth of M. procera. Three pH values (pH 5.0, 5.5 and 6.0), four incubation temperatures (15, 20, 25 and 30°C), seven carbon (C) sources (dextrose, glucose, lactose, maltose, mannitol, sucrose and xylose) and six nitrogen (N) sources ((NH4)2HPO4, NH4NO3 and Ca(NO3)2, malt extract, peptone and yeast extract) were investigated. In the second step of the study, the effect of seven pH values (4.0, 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0) on the mycelial colony diameter was examined at 20 and 25°C since these temperatures gave the best mycelial growth in the previously conducted temperature experiment. The best mycelial growth was determined at pH 6.0. The optimum temperature for mycelial growth of M. procera was found as 25°C. The use of glucose as carbon source and yeast extract and peptone as nitrogen source in the culture medium gave the best results for mycelial growth. Determining of optimum culture conditions for mycelial growth of M. procera will provide important contributions to the fortcoming studies on it’s commercially cultivation in Turkey.

Optimization of Culture Conditions for Growth of Marine Phytoplankton

The present study investigated the optimum levels of ambient temperature, salinity and light intensity for the growth of marine diatoms and microalgae. The marine diatoms Thalassiosira subtilis, Entomoneis paludosa and microalgae Isochrysis galbana were optimized for the growth. Subcultures were prepared in filtered natural seawater enriched with f/2 media for diatoms and Conway media for I.galbana . Cultures were grown under three different levels of three parameters consists of 27 combinations in 96 well plates. Ten replicates of cultures were maintained for each combination of ambient levels. The algal density was determined by spectrometric absorbance of culture at 680 nm. Likewise, the growth was estimated from the rate of increase in the absorbance values over a period of time. The durations of growth differed between the species. The I.galbana was grown for prolonged culture duration up to 15 days followed by E.paludosa with 12 days and T.subtilis with 8 days. Levels of ambient temperature and light intensity are the driving parameters for optimum growth of the species studied while the optimum salinity of 30 psu was observed for all species.Ambient levels of 28 °C, 30 psu and 2500 lux were found optimum for the growth of T. subtilis and E.paludosa attained its optimum growth at 24 °C, 30 psu and 2500 lux. The higher light intensity (4500 lux) hasenhanced the growth of I.galbana along with 24 °C of temperature and 30 psu of salinity.

Thickness-wise growth technique for human articular chondrocytes to fabricate three-dimensional cartilage grafts.

IntroductionCutting the cost of manufacturing is important for extending the use of tissue-engineered therapeutic products. The present study aimed to develop a simple method for fabrication of cartilaginous tissues for regenerative therapy, utilizing the phenomenon where human articular chondrocytes grow thickness-wise and spontaneously form three-dimensionally thick tissues.MethodsNormal human articular chondrocytes (NHACs) were cultured with varying concentrations of transforming growth factor beta 1 (TGF-β1) and/or fibroblast growth factor-2 (FGF-2) to optimize the culture condition for thickness-wise growth of chondrocytes. Next, the tissues grown in the optimal condition were subjected to re-differentiation culture in attached and detached states to assess differentiation capacity by evaluating secreted factors, histological analysis, and a gene expression assay.ResultsNHACs grew thickness-wise efficiently in the presence of 1 ng/mL TGF-β1 and 10 ng/mL FGF-2. After two weeks of culture, NHACs grew with 11-fold higher thickness and 16-fold higher cell number compared to cells which were neither treated with TGF-β1 nor with FGF-2. These thickness-wise-grown chondrocytes could be re-differentiated by a differentiation medium according to the increase in melanoma inhibitory activity (MIA) and positive safranin-O staining. Interestingly, the cartilaginous gene expression was considerably different between the attached and detached conditions even in the same culture medium, indicating the necessity of detachment and shrinkage to achieve further differentiation.ConclusionsSpontaneous thickness-wise growth might provide a simple tissue-engineering method for manufacturing cartilaginous 3D tissues.

English

English

High Kanamycin Concentration as Another Stress Factor Additional to Temperature to Increase pDNA Production in E. coli DH5α Batch and Fed-Batch Cultures.

Plasmid DNA (pDNA) vaccines require high supercoiled-pDNA doses (milligrams) to achieve an adequate immune response. Therefore, processes development to obtain high pDNA yields and productivity is crucial. pDNA production is affected by several factors including culture type, medium composition, and growth conditions. We evaluated the effect of kanamycin concentration and temperature on pDNA production, overflow metabolism (organic acids) and metabolic burden (neomycin phosphotransferase II) in batch and fed-batch cultures of Escherichia coli DH5α-pVAX1-NH36. Results indicated that high kanamycin concentration increases the volumetric productivity, volumetric and specific yields of pDNA when batch cultures were carried out at 42 °C, and overflow metabolism reduced but metabolic burden increased. Micrographs taken with a scanning electron microscope (SEM) were analyzed, showing important morphological changes. The high kanamycin concentration (300 mg/L) was evaluated in high cell density culture (50 gDCW/L), which was reached using a fed-batch culture with temperature increase by controlling heating and growth rates. The pDNA volumetric yield and productivity were 759 mg/L and 31.19 mg/L/h, respectively, two-fold greater than the control with a kanamycin concentration of 50 mg/L. A stress-based process simultaneously caused by temperature and high kanamycin concentration can be successfully applied to increase pDNA production.

Optimization of biomass and fatty acid productivity of Desmodesmus intermedius as a promising microalga for biodiesel production

ABSTRACT The present study aimed to optimize the culture conditions for algal growth and lipid production of Desmodesmus intermedius. Different environmental factors including nitrogen content, salinity, kinetin hormone, vitamin B12, and light intensity were applied individually or in combination using Plackett–Burman statistical experimental design. In addition, the impact of optimized conditions on fatty acids profile and biodiesel quality was studied. Results revealed that the maximum lipid and biomass production for individual study optimization was recorded using 125 mg/L sodium nitrate, 12.5 mg/L sodium chloride, 10−7 mol/L kinetin, and 10−2 mg/L vitamin B12, at 4500 lux. In addition, Plackett-Burman model revealed that combination of sodium nitrate (125 mg/L), sodium chloride (25 mg/L), Kinetin hormone (0 mol/L), vitamin B12 (0 mg/L), and Light intensity (4500 Lux) resulted in maximum lipid productivity. The overall enhancements in lipid productivities using Plaktt–Berman model were 20% and 61.6% over individual optimization and control, respectively. Fatty acids profile for D. intermedius recorded the presence of 14 fatty acids. These fatty acids are suitable to produce high-quality biodiesel.

Optimization of culture condition for growth of the Aurantiochytrium sp. shy, isolated from Persian Gulf

In this study, a native microalgae strain with a high capability of DHA production has been isolated and purified from Mangrove shores in the Persian Gulf. Morphological and molecular identification based on 18S rRNA gene showed that the isolated strain belongs to Aurantiochytrium genus. In order to optimize the growth of this strain, various environmental factors were examined, including temperature(15, 20, 25, 30 and 35°C), salinity (10, 30 and 50 g/L) and different concentrations of nitrogen sources of Yeast extract, Peptone, Meat Extract and Monosodium glutamate, each of which being tested with 10 g/L concentrations. The findings show that the optimal growth conditions occur in 25-30°, pH 6 and presence of Meat Extract as a nitrogen source. In these conditions, the cellular biomass weight, fatty acids and DHA amounts were obtained as 7.34, 2.25 and 0.4 g/L, respectively. These results imply that the new isolated strain of Aurantiochytrium sp.shy is suitable for lipid production.

Phenolic acid and flavonoid production in agar, agitated and bioreactor-grown microshoot cultures of Schisandra chinensis cv. Sadova No. 1 – a valuable medicinal plant

In vitro cultures and raw materials (fruits and leaves) of the valuable medicinal plant species – Schisandra chinensis cultivar Sadova No. 1 (SchS) – were evaluated for the production of two groups of phenolic compounds, phenolic acids and flavonoids, and their antioxidant potential. A series of experiments was conducted, aimed at optimizing culture conditions for maximum growth and phenolic production in SchS microshoots. Different concentrations of plant growth regulators (6-benzyladenine – BA and 1-naphthaleneacetic acid – NAA, from 0 to 3 mg/l) in Murashige-Skoog (MS) medium were tested in several cultivation systems (agar, agitated, bioreactor) over various growth periods (10, 20, 30, 40, 50 and 60 days). Furthermore, an elicitation experiment was conducted in which the bioreactor-grown microshoots were exposed to yeast extract. HPLC-DAD analyses confirmed the presence of eight phenolic acids – chlorogenic, cryptochlorogenic, gallic, neochlorogenic, protocatechuic, salicylic, syringic and vanillic, and two flavonoids: kaempferol and quercitrin, in the in vitro biomasses. The highest total phenolic acid (357.93 mg/100 g DW) and flavonoid (105.07 mg/100 g DW) contents were obtained in agar culture extracts cultivated for 30 days on MS medium containing 2 mg/l BA and 0.5 mg/l NAA and for 50 days on MS medium containing 0.1 mg/l BA and 2 mg/l NAA, respectively. These amounts were 1.59- and 5.95-fold lower than in parent plant leaf extracts (569.66 mg/100 g DW), and 4.30- and 1.25-fold higher than in fruit extracts (83.17 mg/100 g DW), respectively. Microshoots grown in a Plantform bioreactor also proved to be a good source of phenolic compounds, however, the elicitor treatment had no noticeable effect on their accumulation. Antioxidant capacity assessed by the Folin-Ciocalteu, FRAP, DPPH and CUPRAC assays revealed significantly higher potential in extracts from in vitro biomass and leaves of the parent plant, as compared to the parent plant fruit extracts.