Optimal Culture Conditions Research Articles

Characterization of Commercially Available Human Primary Alveolar Epithelial Cells.

In vitro lung research requires appropriate cell culture models that adequately mimic invivo structure and function. Previously, researchers extensively used commercially available and easily expandable A549 and NCI-H441 cells, which replicate some but not all features of alveolar epithelial cells. Specifically, these cells are often restricted by terminally altered expression while lacking important alveolar epithelial characteristics. Of late, human primary alveolar epithelial cells (hPAEpCs) have become commercially available but are so far poorly specified. Here, we applied a comprehensive set of technologies to characterize their morphology, surface marker expression, transcriptomic profile, and functional properties. At optimized seeding numbers of 7,500 cells per square centimeter and growth at a gas-liquid interface, hPAEpCs formed regular monolayers with tight junctions and amiloride-sensitive transepithelial ion transport. Electron microscopy revealed lamellar body and microvilli formation characteristic for alveolar type II cells. Protein and single-cell transcriptomic analyses revealed expression of alveolar type I and type II cell markers; yet, transcriptomic data failed to detect NKX2-1, an important transcriptional regulator of alveolar cell differentiation. With increasing passage number, hPAEpCs transdifferentiated toward alveolar-basal intermediates characterized as SFTPC-, KRT8high, and KRT5- cells. In spite of marked changes in the transcriptome as a function of passaging, Uniform Manifold Approximation and Projection plots did not reveal major shifts in cell clusters, and epithelial permeability was unaffected. The present work delineates optimized culture conditions, cellular characteristics, and functional properties of commercially available hPAEpCs. hPAEpCs may provide a useful model system for studies on drug delivery, barrier function, and transepithelial ion transport invitro.

Optimization of physicochemical parameters for lipopeptides surfactin production by Bacillus halotolerans under oat straw medium using Box-Behnken design and artificial neural networks

The primary objective of this study was to produce lipopeptide surfactin using Bacillus halotolerans (GenBank accession: ON204239), which was isolated from the rhizosphere of pine trees in the El Baaraouia forest in Eastern Algeria. B. halotolerans demonstrated the capacity to biosynthesize surfactants when cultured on a blood agar medium, resulting in the formation of clear zones around the colonies. Antagonism tests were conducted using the culture supernatant on two liquid media: LB (Luria-Bertani) and a natural medium (OSM) with oats as the sole carbon source. The results showed significant inhibition zones (15 mm) on LB medium compared to larger zones (25 mm) on OSM after 24 h. In vivo, antagonistic activity under field conditions after 12 weeks revealed that B. halotolerans inoculated in OSM effectively enhanced and protected tomato plants against F. napiforme compared to the LB medium and the control group. The results exhibited notable differences in plant size, number of leaves, number of discolored leaves, and root mass, all favoring the OSM-inoculated plants. The study utilized a combination of Box-Behnken design within the response surface methodology (BBD-RSM), artificial neural networks (ANNs), and the genetic algorithm (GA) to optimize culture conditions and achieve maximum surfactin production. The optimal conditions for surfactin production included a substrate concentration of 7 g, incubation time of 30 h, pH of 2.31, and a temperature of 37.4 °C. Under these conditions, the antagonism test against phytopathogenic F. napiforme (GenBank accession: ON204349) using the culture supernatant revealed a significant inhibition zone of 18.5 mm after 24 h. UPLC-MS analysis of the lipopeptides produced by B. halotolerans cultured on an oat straw medium confirmed the production of various surfactin analogs. Seven analogs (C12, C13, C14, C15, C16, C17, and C18) were identified. Notably, the production of surfactin analogs by B. halotolerans in the Luria-Bertani medium yielded fewer analogs (C12, C13, C14, C15, and C16) in comparison. In summary, this study successfully optimized surfactin production conditions using BBD-RSM, ANNs, and GA, leading to enhanced antagonistic activity against F. napiforme and the identification of multiple surfactin analogs.

Decellularized fetal collagen exhibits chondroinductive potential for bone marrow-derived mesenchymal stem cells by enhancing glycosaminoglycan production

Articular cartilage repair is challenging due to limited access to reparative cells and a lack of self-healing mechanisms. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are a promising therapeutic option, but their tendency to form fibrocartilage during repair necessitates the optimization of culture conditions. To overcome this limitation, optimizing in-vitro culture conditions with biological coating using extracellular matrix-derived proteins has been efficient in mimicking in-vivo cellular behavior. Fetal cartilage, with abundant collagen, proteoglycans and glycosaminoglycans has emerged as a potential source for cartilage repair. No studies have so far evaluated the effect of fetal cartilage-derived collagen on BM-MSCs. This study aimed to evaluate the chondro-inductive potential of decellularized collagen derived from fetal cartilage, which was used as a coating material for expansion of BM-MSCs. The extraction of fetal collagen was performed from the tibiofemoral joint of a 36+4-week gestational age fetus. The freeze-dried collagen type II was reconstituted at a concentration of 10μg/ml and used to coat the culture flasks. Passage 3 BM-MSCs were divided into two groups: a) standard expansion medium (BM-MSCs) and b) collagen-coated plasticware (collagen-coated BM-MSCs). Growth kinetics, surface markers, gene expression, and differentiation potential were assessed. The decellularized collagen coating did not influence the growth kinetics, surface marker and gene expression of BM-MSCs. However, it positively influenced GAG accumulation and collagen type II deposition. Further studies utilizing in-vivo models are warranted to evaluate the potential of collagen-coated BM-MSCs and exploit their adjuvant effect on chondrogenesis.

Kinetic analysis and mechanism of nitrate, calcium, and cadmium removal using the newly isolated Pseudomonas sp. LYF26

The co-existence of heavy metals and nitrate (NO3−-N) pollutants in wastewater has been a persistent global concern for a long time. A strain LYF26, which can remove NO3−-N, calcium (Ca(II)), and cadmium (Cd(II)) simultaneously, was isolated to explore the properties and mechanisms of synergistic contaminants removal. Different conditions (Cd(II) and Ca(II) concentrations and pH) were optimized by Zero-, Half-, and First-order kinetic analyses to explore the environmental parameters for the optimal effect of strain LYF26. Results of the kinetic analyses revealed that the optimal culture conditions for strain LYF26 were pH of 6.5, Cd(II) and Ca(II) concentrations of 3.00 and 180.00 mg L−1, accompanied by Ca(II), Cd(II), and NO3−-N efficiencies of 53.10%, 90.03%, and 91.45%, respectively. The removal mechanisms of Cd(II) using strain LYF26 as a nucleation template were identified as biomineralization, lattice substitution, and co-precipitation. The differences and changes of dissolved organic matter during metabolism were analyzed and the results demonstrated that besides the involvement of extracellular polymeric substances in the precipitation of Cd(II) and Ca(II), the high content of humic acid-like species revealed a remarkable contribution to the denitrification process. This study is hopeful to contribute a theory for further developing microbially induced calcium precipitation used to treat complex polluted wastewater.

Enhancing polyethylene degradation: a novel bioprocess approach using Acinetobacter nosocomialis pseudo-resting cells.

Despite the discovery of several bacteria capable of interacting with polymers, the activity of the natural bacterial isolates is limited. Furthermore, there is a lack of knowledge regarding the development of bioprocesses for polyethylene (PE) degradation. Here, we report a bioprocess using pseudo-resting cells for efficient degradation of PE. The bacterial strain Acinetobacter nosocomialis was isolated from PE-containing landfills and characterized using low-density PE (LDPE) surface oxidation when incubated with LDPE. We optimized culture conditions to generate catalytic pseudo-resting cells of A. nosocomialis that are capable of degrading LDPE films in a bioreactor. After 28days of bioreactor operation using pseudo-resting cells of A. nosocomialis, we observed the formation of holes on the PE film (39 holes per 217 cm2, a maximum diameter of 1440μm). This study highlights the potential of bacteria as biocatalysts for the development of PE degradation processes. KEY POINTS: • New bioprocess has been proposed to degrade polyethylene (PE). • Process with pseudo-resting cells results in the formation of holes in PE film. • We demonstrated PE degradation using A. nosocomialis as a biocatalyst.

IoAT: Internet of Aquaculture Things for Monitoring Water Temperature in Tiger Shrimp Ponds with DS18B20 Sensors and WeMos D1 R2

Cultivation of tiger prawns stands as a crucial sector in Indonesia's fisheries industry, significantly contributing to the country's foreign exchange. However, challenges persist in the cultivation process, particularly concerning suboptimal harvest outcomes. A critical factor in tiger prawn cultivation is the water temperature within shrimp ponds, a parameter directly influencing shrimp growth. The recommended normal temperature range is 28-31°C. Deviations from this range can adversely impact the shrimp's metabolic system and appetite, resulting in stress and potential mortality. Temperature fluctuations can lead to severe issues such as hindered growth, reduced productivity, and increased shrimp mortality. Real-time monitoring of air temperature emerges as a pivotal element in ensuring the success of shrimp farming. This research aims to provide a practical solution for shrimp cultivation by presenting a system that enables farmers to adjust air temperature in ponds in real-time through a user-friendly website application. The ability to promptly respond to abnormal temperature fluctuations empowers farmers to optimize cultivation conditions, thereby reducing shrimp mortality rates. The research focuses on creating a water temperature monitoring system for tiger prawn ponds using cloud storage through the Firebase platform. By implementing real-time temperature monitoring, financial risks for shrimp farmers can be mitigated, preventing losses attributed to temperature-induced shrimp mortality. The research utilizes the DS18B20 temperature sensor and WeMos D1 R2 as the control center. The website displays air temperature measurements, showcasing a high accuracy of 99% with a minimal error of 1.2%. This underscores the system's effectiveness in measuring air temperature both above and below the pond. The incorporation of IoT technology for monitoring water quality in ponds offers a practical and innovative approach to tiger prawn cultivation, with the potential to enhance production outcomes in each harvest.

Optimization of bacterioruberin production from Halorubrum ruber and assessment of its antioxidant potential

Haloarchaea produce bacterioruberin, a major C50 carotenoid with antioxidant properties that allow for its potential application in the food, cosmetic, and pharmaceutical industries. This study aimed to optimize culture conditions for total carotenoid, predominantly comprising bacterioruberin, production using Halorubrum ruber MBLA0099. A one-factor-at-a-time and statistically-based experimental design were applied to optimize the culture conditions. Culture in the optimized medium caused an increase in total carotenoid production from 0.496 to 1.966 mg L− 1 Maximal carotenoid productivity was achieved in a 7-L laboratory-scale fermentation and represented a 6.05-fold increase (0.492 mg L–1 d–1). The carotenoid extracts from strain MBLA0099 exhibited a 1.8–10.3-fold higher antioxidant activity in vitro, and allowed for a higher survival rate of Caenorhabditis elegans under oxidative stress conditions. These results demonstrated that Hrr. ruber MBLA0099 has significant potential as a haloarchaon for the commercial production of bacterioruberin.

Screening, characterization, and production of Bacillus cereus (S55) bioflocculant isolated from soil for application in wastewater treatment

This research evaluated a bacterial strain, Bacillus cereus (S55), isolated from the soil for its bioflocculant production capabilities. The strain was identified through morphological and 16s rRNA gene sequencing. The optimization of culture conditions, using One-factor-at-a-time method, significantly enhanced bioflocculant production with glucose and urea. FTIR analysis showed the presence of hydroxyl, amine and carboxylate functional groups, with polysaccharides as predominant components. Scanning electron microscopy and X-ray diffraction confirmed the crystalline nature of the bioflocculant. The strain studied showed potential in treating household wastewater and was effective at removing dyes, suggesting alternatives for wastewater management.

Insights into the synthesis, engineering, and functions of microbial pigments in Deinococcus bacteria.

The ability of Deinococcus bacteria to survive in harsh environments, such as high radiation, extreme temperature, and dryness, is mainly attributed to the generation of unique pigments, especially carotenoids. Although the limited number of natural pigments produced by these bacteria restricts their industrial potential, metabolic engineering and synthetic biology can significantly increase pigment yield and expand their application prospects. In this study, we review the properties, biosynthetic pathways, and functions of key enzymes and genes related to these pigments and explore strategies for improving pigment production through gene editing and optimization of culture conditions. Additionally, studies have highlighted the unique role of these pigments in antioxidant activity and radiation resistance, particularly emphasizing the critical functions of deinoxanthin in D. radiodurans. In the future, Deinococcus bacterial pigments will have broad application prospects in the food industry, drug production, and space exploration, where they can serve as radiation indicators and natural antioxidants to protect astronauts' health during long-term space flights.

Potential of Microalgae in Bioethanol Production and Optimization of Cultivation Conditions

Potential of Microalgae in Bioethanol Production and Optimization of Cultivation Conditions

Increasing sustainability and reproducibility of in vitro toxicology applications: serum-free cultivation of HepG2 cells.

Fetal Bovine Serum (FBS) is an important ingredient in cell culture media and the current standard for most cells in vitro. However, the use of FBS is controversial for several reasons, including ethical concerns, political, and societal pressure, as well as scientific problems due to the undefined and variable nature of FBS. Nevertheless, scientists hesitate to change the paradigm without solid data de-risking the switch of their assays to alternatives. In this study, HepG2 cells, a human hepatoblastoma cell line commonly used to study drug hepatotoxicity, were adapted to serum-free conditions by using different commercially available media and FBS replacements. After transition to these new culture conditions, the success of adaptation was determined based on cell morphology and growth characteristics. Long-term culturing capacity for each medium was defined as the number of passages HepG2 cells could be cultured without any alterations in morphology or growth behavior. Two media (Advanced DMEM/F12 from ThermoFisher and TCM® Serum Replacement from MP Biomedicals) showed a long-term cultivation capacity comparable to media containing FBS and were selected for further analysis. Both media can be characterized as serum-free, however still contain animal-derived components: bovine serum albumin (both media) and bovine transferrin (only TCM® serum replacement). To assess the functionality of the cells cultivated in either of the two media, HepG2 cells were treated with reference compounds, specifically selected for their known hepatotoxicity characteristics in man. Different toxicological assays focusing on viability, mitochondrial toxicity, oxidative stress, and intracellular drug response were performed. Throughout the different assays, response to reference compounds was comparable, with a slightly higher sensitivity of serum-free cultivated HepG2 cells when assessing viability/cell death and a lower sensitivity towards oxidative stress. Taken together, the two selected media were shown to support growth, morphology, and function of serum-free cultivated HepG2 cells in the early preclinical safety space. Therefore, these results can serve as a starting point to further optimize culture conditions with the goal to remove any remaining animal-derived components.

A Metabolic Perspective on Nitrogen Source and Temperature in Microalgae

The roles of nitrogen (N) and temperature in primary producer metabolism are becoming increasingly relevant for optimising algal culturing and for understanding how global warming and changing agricultural practices will influence crops. I argue that these factors could be combined into a single mechanistic framework based on the 'metabolic theory of ecology (MTE)', which has been successful in other fields. The hallmark of this approach is the lower temperature sensitivity of photosynthesis (P) compared with respiration (R), which reduces carbon allocation to biomass (carbon-use efficiency [CUE]) with warming. Notably, it has been reported that photorespiration associated with N assimilation could explain reduced thermal sensitivity of P in C3 organisms. If true, this means that the N source should change the thermal sensitivity of P and therefore shape the thermal responses of CUE and growth, providing a simple mechanistic framework for predicting the impacts of N and temperature on C3 organisms. To test this metabolic perspective, I cultured the tractable green alga, <i>Chlorella vulgaris </i>Beijer., and measured the rates of P and R alongside the rates of CUE and growth when provided with inorganic (nitrate ions or ammonium) or organic (glutamine) N sources across a temperature gradient. As expected, organic N increased the thermal sensitivity of P but not R, with an associated increase in CUE with warming. This work suggests that we could give this MTE-based perspective the 'green light' to inform optimal algal culturing conditions in industrial applications, and further work should investigate the possible impacts of agricultural organic N use in a warmer world.

Optimization of culture condition for Spodoptera frugiperda by design of experiment approach and evaluation of its effect on the expression of hemagglutinin protein of influenza virus.

The baculovirus expression vector system (BEVS) is a powerful tool in pharmaceutical biotechnology to infect insect cells and produce the recombinant proteins of interest. It has been well documented that optimizing the culture condition and its supplementation through designed experiments is critical for maximum protein production. In this study, besides physicochemical parameters including incubation temperature, cell count of infection, multiplicity of infection, and feeding percentage, potential supplementary factors such as cholesterol, polyamine, galactose, pluronic-F68, glucose, L-glutamine, and ZnSO4 were screened for Spodoptera frugiperda (Sf9) cell culture and expression of hemagglutinin (HA) protein of Influenza virus via Placket-Burman design and then optimized through Box-Behnken approach. The optimized conditions were then applied for scale-up culture and the expressed r-HA protein was characterized. Optimization of selected parameters via the Box-Behnken approach indicated that feed percentage, cell count, and multiplicity of infection are the main parameters affecting r-HA expression level and potency compared to the previously established culture condition. This study demonstrated the effectiveness of designing experiments to select and optimize important parameters that potentially affect Sf9 cell culture, r-HA expression, and its potency in the BEVS system.

Optimization of culture conditions for endophytic bacteria in mangrove plants and isolation and identification of bacteriocin.

Introduction: The antibacterial protein PAG14 was extracted from a metabolite of Bacillus G14 isolated from mangrove plants. Methods: In this study, Pseudomonas aeruginosa, Pasteurell multocide, Enterobacter aerogenes, and Enterococcus faecalis were used as indicator bacteria to screen endophytes that exhibited antibacterial activity. The endophyte culture conditions were optimized to enhance productivity. Subsequently, the culture supernatant was salted using ammonium sulfate, followed by purification using dextran gel chromatography and ion exchange column techniques. Finally, the structures of antibacterial proteins were identified using mass spectrometry. Results and Discussion: The optimal culture conditions for Bacillus G14 were 2% mannitol, 0.5% fish peptone, 0.05% KH2PO4 + 0.05% K2HPO4 + 0.025% MnSO4·H2O. The antibacterial substances exhibited stability within the temperature range of 30-40°C and pH range of 5.0-7.0, while displaying sensitivity toward enzymes. The antibacterial activity decreased as the duration of UV irradiation increased. The antibacterial protein PAG14, isolated from the culture broth of Bacillus G14 through purification using dextran gel and ion-exchange columns, was identified as a class III bacteriocin using LC-MS/MS, similar to Lysozyme C. These findings serve as a theoretical foundation for the investigation and application of bacteriocins in food products.

Optimization of culture conditions of Scenedesmus sp. algae and catalytic performance of a NiFe2O4@SiO2/MgO magnetic nano-catalyst for biodiesel production.

This study aims to optimize the lipid content of Scenedesmus sp. for high-yield biodiesel production. Three factors affecting the culture conditions, namely salinity, nitrogen concentration, and light intensity, were selected and their effects on the maximum lipid content were investigated using the Box Behnken design. The results showed that the maximum lipid content (32.7% of algal dry weight) was obtained in the algal samples cultured under the optimized conditions. A core-shell magnetic nano-catalyst, NiFe2O4@SiO2/MgO, was synthesized and used to produce biodiesel via the transesterification reaction. The nano-catalyst was characterized by field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray powder diffraction analysis (XRD), vibrating sample magnetometry (VSM), elemental mapping techniques, transmission electron microscopy (TEM), and Fourier transform infrared spectroscopy (FT-IR). Using the Box Behnken design and keeping the temperature constant, the molar ratio of methanol to oil, the amount of catalyst, and the time were optimized to achieve the maximum yield of biodiesel. The maximum yield of biodiesel was 95.3% under the optimal conditions.

Obtaining a primary suspension cell culture of Dracocephalum palmatum Stephan ex Willd

Dracocephalum palmatum Steph. grows on the southern slopes of the Oymyakon plateau in Yakutia (Northeast of Russian Federation) in conditions of harsh continental climate with continuous permafrost. The aboveground phytomass of the plant contains various complexes of secondary metabolites including polyphenolic compounds. It is a potential source of secondary metabolites needed for practical use in the pharmaceutical industry. The aim of the study is to obtain a primary suspension cell culture of Dracocephalum palmatum, growing in the conditions of the Cold Pole — Oymyakon. The work includes optimization of the nutrient medium for introducing calluses into a suspension culture, analysis of the dynamics of biomass growth of the obtained suspension culture, and morphological characteristics of the cells of the suspension culture. The callus cell cultures of Dracocephalum palmatum, cultivated on Murashige-Skoog (MS) medium with the addition of 0.5 mg/L α- naphthylacetic acid (NAA) and 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), were most successfully transitioned into suspension culture. The maximum biomass growth of cell suspension culture was observed when cultivated in liquid MS medium with the addition of 2,4-D (0.5 mg/L), 6- benzylaminopurine (0.5 mg/L), and NAA (0.5 mg/L). The primary cell suspension culture of Dracocephalum palmatum, cultivated for 22 days, had an increase in wet weight of 9,2084 g, dry weight — 0,34135 g, and contained dedifferentiated aggregates of parenchyma-like cells and single round-shaped cells. Samples of the obtained cell suspension culture of Dracocephalum palmatum will be used for the analysis of secondary metabolites and for the development of optimal cultivation conditions in a bioreactor.

The Reaction of Some Maize Hybrids to Several Plant Densities, in the Cultivation Conditions of West Part of Romania

The density of plants on a certain area is a method used on a large scale to increase yield, but in certain environmental conditions and cultivation areas, it can be a stress for plants. At the Agricultural Research and Development Station Turda (ARDS Turda), seven maize hybrids, the most recent creations, were tested at four plant densities, in order to monitor their reaction in non-irrigated conditions. The plant densities used were 60,000, 70,000, 80,000 and 90,000 plant/ha and the experiment was carried out in three years: 2020, 2021 and 2022. The behaviour of the hybrids was different, but most of them had the best yield at 60,000 plants/ha. However, two hybrids stood out, Turda 335 and Turda 380, which had very good yield at 70,000 plants/ha, as well as a better tolerance to the stress of higher densities, especially due to characteristics such as superior production capacity, erect port or semi-erect leaves, prolific, good tolerance to water stress. Especially in non-irrigated conditions, maize yield is highly influenced by climatic conditions, regardless of density, so the use of high densities must be associated with ensuring optimal cultivation conditions.

Optimizing cultivation conditions for enhanced productivity Limnothrix planctonica through pH variation and light quality

The survival and growth of cyanobacteria in diverse environments are influenced by several critical factors when cultivating them for research or industrial purposes. Ensuring their optimal growth necessitates consideration of these factors that potentially impede pigment and biomass production. Therefore, optimizing light conditions and pH levels enhances biomass and pigment yield productivity in cyanobacterial cultivation. This study investigates the cultivation conditions of Limnothrix planctonica (KU.B3) in a BG-11 liquid medium, focusing on pigment production, growth response to light and pH variations, and photosynthetic efficiency assessment. Results demonstrated efficient growth of L. planctonica (KU.B3) under white, orange, and red light within the pH range of 7.4–9.0. Cultivation at pH 7.4, yielded the highest biomass (3.59 ± 0.35 μg ml−1) and phycocyanin content (0.016 ± 0.01 mg g−1 dry weight). Red light-induced significant phycocyanin yield (5.49 ± 1.21 mg g−1 dry weight), while white light promoted substantial biomass production (10.03 ± 1.28 μg ml−1). Conversely, exposure to blue light reduced biomass but exhibited the highest photosynthetic efficiency (Fv / Fm = 0.48 ± 0.06). The study underscores the tradeoff between productivity and photosynthetic efficiency, emphasizing the need for further research to optimize conditions for both parameters, especially for industrial-scale applications.

Bovine embryo production in vitro: evolution of culture media and commercial perspectives.

In vitro produced embryos exhibit lower viability compared to their in vivo counterparts. Mammalian preimplantation embryos have the ability to reach the blastocyst stage in diverse culture media, showcasing considerable metabolic adaptability, which complicates the identification of optimal developmental conditions. Despite embryos successfully progressing to the blastocyst stage, adaptation to suboptimal culture environments may jeopardize blastocyst viability, cryotolerance, and implantation potential. Enhancing our capacity to support preimplantation embryonic development in vitro requires a deeper understanding of fundamental embryo physiology, including preferred metabolic substrates and pathways utilized by high-quality embryos. Armed with this knowledge, it becomes achievable to optimize culture conditions to support normal, in vivo-like embryo physiology, mitigate adaptive stress, and enhance viability. The objective of this review is to summarize the evolution of culture media for bovine embryos, highlighting significant milestones and remaining challenges.

Технология культивирования Limnospira fusiformis из озера Соленого (г. Омск)

Global food shortages make it necessary to look for alternative renewable bioresources. In the south of Western Siberia, the filamentous cyanoprokaryote Limnospira fusiformis triggers seasonal algae bloom in hypergaline alkaline Lake Solenoye. The species has valuable nutritional properties, and its phytomass is a potential source of proteins and biologically active substances. The O9.13F strain of L. fusiformis has a good potential as a bioadditive in animal feeding. The article offers a technology for cultivating O9.13F of L. fusiformis, isolated from Lake Solenoye. The research objectives were to establish the optimal cultivation conditions, medium, and periodicity.
 The study featured strain O9.13F of filamentous cyanoprokaryote L. fusiformis. The micropipette method made it possible to isolate pure culture from water samples taken from Lake Solenoye, Omsk, Russia, at the end of algae bloom. The cultivation involved a UT-6070 climatic chamber under uniform illumination with light intensity 10–30 μmol photons/m2s and 12-h light-dark circle at 20 ± 2°C on various media: natural habitat – water from the Solenoye Lake; mineral medium – liquid Zarrouk’s medium; agarized Zarrouk’s medium; composite variants, where the ratio of mineral medium vs. water varied from 1:9 to 9:1.
 Lake water inhibited the culture growth: the trichomes died and sank on day 10–15. Zarrouk’s agarized medium stopped the culture growth as early as on day 2. The most intensive growth and development of the culture was observed in the samples with Zarrouk’s mineral liquid medium and a composite mix of Zarrouk’s medium and sterilized water at a ratio of 5:5. Without stirring, full-fledged trichomes had no time to develop, and the increase in phytomass volume slowed down. O9.13F showed the highest rate of phytomass growth at a cultivation temperature of 20 ± 2°C and a light intensity of 10–30 μmol photons/m2s. The recommended light-dark circle was12:12 h. Zarrouk’s mineral liquid medium and a composite medium of Zarrouk’s medium and sterilized water proved to be optimal in a UT-6070 environmental chamber. Europolitext KV-06 or mechanical mixing could prevent sedimentation of phosphates after 20 days of cultivation. The optimal recultivation frequency was once every 5–7 days. 
 The new cultivation technology made it possible to obtain a significant volume of L. fusiformis phytomass in a short time and with low financial expenses.