• MeSH
• Bacteria enzymologie   růst a vývoj   metabolismus   MeSH
• bibliografie jako téma * MeSH
• Eukaryota enzymologie   růst a vývoj   metabolismus   MeSH
• houby enzymologie   růst a vývoj   metabolismus   MeSH
• kultivace virů MeSH
• kultivační média MeSH
• mikrobiální genetika MeSH
• teoretické modely MeSH
• viry enzymologie   růst a vývoj   metabolismus   MeSH
• zdravotnické prostředky MeSH
• Publikační typ
• bibliografie MeSH
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• kultivační média MeSH

In vitro culturability of Murman strain of Tuleniy flavivirus isolated recently in the northern regions of the USSR was studied. Stable PS pig kidney line was found suitable as a primary sensitive cell substrate for the isolation, proliferation and serial propagation of the virus. The pronounced pathogenicity of the virus to PS cells permits the testing of its infective activity comparable with i.c. titrations on mice, VNT in vitro and the plaquing technique. PS line is suitable for the demonstration and identification of the virus antigen and/or for the study of reproduction of the virus on cellular level using the technique of immunofluorescence.

• MeSH
• antigeny virové analýza   MeSH
• arboviry růst a vývoj   imunologie   MeSH
• druhová specificita MeSH
• fluorescenční protilátková technika MeSH
• kultivace virů * MeSH
• lidé MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny virové MeSH

In terms of the number and diversity of living units, the prokaryotic empire is the most represented form of life on Earth, and yet it is still to a significant degree shrouded in darkness. This microbial "dark matter" hides a great deal of potential in terms of phylogenetically or metabolically diverse microorganisms, and thus it is important to acquire them in pure culture. However, do we know what microorganisms really need for their growth, and what the obstacles are to the cultivation of previously unidentified taxa? Here we review common and sometimes unexpected requirements of environmental microorganisms, especially soil-harbored bacteria, needed for their replication and cultivation. These requirements include resuscitation stimuli, physical and chemical factors aiding cultivation, growth factors, and co-cultivation in a laboratory and natural microbial neighborhood.

• Klíčová slova
• VBNC, cultivation techniques, difficult-to-culture microorganisms, dormancy, environmental microbiome, growth factors, improved cultivation, microbial ecology,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Monoxenous (one host) trypanosomatids from insects and other invertebrates can be introduced into axenic culture relatively easily and efficiently, allowing for their transfer from the field into the laboratory. Here we describe simple methods and alternative cultivation protocols, the wider application of which will allow substantial expansion of trypanosomatids available for research.

• Klíčová slova
• Axenization, Cultivation, Field, Insects, Isolation, Trypanosoma, Trypanosomatids,
• MeSH
• axenická kultura metody   MeSH
• hmyz parazitologie   MeSH
• parazitologie přístrojové vybavení   metody   MeSH
• Trypanosomatina izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Circulating tumor cells (CTCs) are important targets for treatment and critical surrogate markers when evaluating cancer prognosis and therapeutic response. A sensitive methodology for detecting CTCs in gastric cancer (GC) patients is needed. In this study we demonstrate a device for enrichment and cultivation of CTCs. In total, 22 patients with GC, all candidates for surgery, were enrolled in the study. Peripheral blood samples were collected before surgery, and patients were re-evaluated within operation and divided into two groups: resectable and non-resectable GC. A new size-based separation test for enrichment and cultivation of CTCs was used (MetaCell(®)). In addition to cytomorphological analysis, gene expression of tumor associated genes (Cytokeratin-18, Cytokeratin-19, Cytokeratin-20, Cytokeratin-7, EPCAM, MUC1, HER2, EGFR) and of leukocyte markers (e.g. CD45, CD68) was tested in enriched CTC fractions. CTCs were detected in 59 % of the patients studied (n = 13/22). CTCs were detected in seven patients of the resection group (7/10, 70 %) and six of the non-resectable group (6/12, 50 %). Enrichment of the viable CTCs allowed subsequent successful cultivation in vitro. The cytomorphological characterization of the CTCs was a prerequisite of random gene expression testing in CTC-positive samples. In CTC-positive samples gene expression of cytokeratin 18 and 19 was elevated in comparison to the whole blood gene expression analysis. CTCs were found to be present in both resectable and non-resectable gastric cancer patients. The size-based separation platform for CTCs may be used for in vitro cultivation, as well as in subsequent molecular analysis if desired. The sensitivity of CTC-detection could be enhanced by the combination of cytomorphological and molecular analysis.

• Klíčová slova
• CTC, Circulating tumor cells, Cultivation, Gastric cancer, Metacell,
• Publikační typ
• časopisecké články MeSH

• MeSH
• adsorpce MeSH
• cytopatogenní efekt virový MeSH
• hemaglutinační testy MeSH
• Hepatovirus * MeSH
• kultivace virů MeSH
• lidé MeSH
• neutralizační testy MeSH
• Orthomyxoviridae MeSH
• techniky in vitro MeSH
• tvorba protilátek MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The cultivation and investigation of strictly anaerobic microorganisms belong to the fields of anaerobic microbial physiology, microbiology, and biotechnology. Anaerobic cultivation methods differ from classic microbiological techniques in several aspects. The requirement for special instruments, which are designed to prevent the contact of the specimen with air/molecular oxygen by different means of manipulation, makes this field more challenging for general research compared to working with aerobic microorganisms. Anaerobic microbiological methods are required for many purposes, such as for the isolation and characterization of new species and their physiological examination, as well as for anaerobic biotechnological applications or medical indications. This review presents the historical development of methods for the cultivation of strictly anaerobic microorganisms focusing on methanogenic archaea, anaerobic cultivation methods that are still widely used today, novel methods for anaerobic cultivation, and almost forgotten, but still relevant, techniques.

• Klíčová slova
• anaerobes, biogas, cultivation methods, methane, methanogens,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

BACKGROUND: The extreme conditions of thermal springs constitute a unique aquatic habitat characterized by low nutrient contents and the absence of human impacts on the microbial community composition. Thus, these springs may host phylogenetically novel microorganisms with potential use in biotechnology. With this hypothesis in mind, we examined the microbial composition of four thermal springs of the world-renowned spa town of Karlovy Vary (Carlsbad), Czechia, which differ in their temperature and chemical composition. RESULTS: Microbial profiling using 16S rRNA gene sequencing revealed the presence of phylogenetically novel taxa at various taxonomic levels, spanning from genera to phyla. Many sequences belonged to novel classes within the phyla Hydrothermae, Altiarchaeota, Verrucomicrobia, and TA06. Cultivation-based methods employing oligotrophic media resulted in the isolation of 44 unique bacterial isolates. These include strains that withstand concentrations of up to 12% NaClw/v in cultivation media or survive a temperature of 100 °C, as well as hitherto uncultured bacterial species belonging to the genera Thermomonas, Paenibacillus, and Cellulomonas. These isolates harbored stress response genes that allow them to thrive in the extreme environment of thermal springs. CONCLUSIONS: Our study is the first to analyze the overall microbial community composition of the renowned Karlovy Vary thermal springs. We provide insight into yet another level of uniqueness of these springs. In addition to their unique health benefits and cultural significance, we demonstrate that these springs harbor phylogenetically distinct microorganisms with unusual life strategies. Our findings open up avenues for future research with the promise of a deeper understanding of the metabolic potential of these microorganisms.

• Klíčová slova
• Amplicon sequencing analysis, Cultivation analysis, Phylogenetic novelty, Thermal water springs,
• Publikační typ
• časopisecké články MeSH

UNLABELLED: Malignant pleural mesothelioma (MPM) is an aggressive disease with very poor prognosis which tends to affect older patients. Progress in the management of this group of patients has been limited by the rarity of the disease and hence, difficulty in conducting randomized trials. The vast majority of cancer deaths occur due to metastasis of the primary tumor to distant sites via circulating tumor cells (CTCs) in the circulation. CTCs are extremely rare and limits in technology used to capture these cells hamper our complete understanding over the metastatic process. In the present study we present a new method for detection and cultivation of CTCs isolated from peripheral blood of MPM patients. PATIENTS AND METHODS: Patients with diagnosed MPM were enrolled into this study. RESULTS: A size-based separation method for viable CTC enrichment from unclothed peripheral blood has been introduced; MetaCell. The size-based enrichment process was based on filtration of peripheral blood (PB) through porous polycarbonate membrane. The separated CTCs are cultured on the membrane in vitro under standard cancer cell culture conditions and observed by an inverted microscope. CONCLUSION: The reported methodology allows for quick and easy enrichment of CTCs and their cultivation. The cultivated cells can be used for next specification of gene expression and histological/biological specificity of concrete mesothelioma.

• Klíčová slova
• CTC, Mesothelioma, MetaCell, circulating tumor cells, cultivation,
• MeSH
• lidé středního věku MeSH
• lidé MeSH
• maligní mezoteliom MeSH
• mezoteliom krev   patologie   MeSH
• nádorové cirkulující buňky patologie   MeSH
• nádory pleury krev   patologie   MeSH
• nádory plic krev   patologie   MeSH
• senioři MeSH
• separace buněk metody   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Leukemia is driven by complex interactions within the inherently hypoxic bone marrow microenvironment, impacting both disease progression and therapeutic resistance. Co-cultivation of leukemic cells with feeder cells has emerged as a valuable tool to mimic the bone marrow niche. This study explores the interplay between human commercial SD-1 and patient-derived UPF26K leukemic cell lines with feeders - human fibroblasts (NHDF) and mesenchymal stem cells (hMSCs) under normoxic and hypoxic conditions. RESULTS: Co-cultivation with feeders significantly enhances proliferation and glycolytic activity in the SD-1 cells, improving their viability, while this interaction inhibits the growth and glucose metabolism of the feeders, particularly NHDF. In contrast, UPF26K cells show reduced proliferation when co-cultivated with the feeders while this interaction stimulates NHDF and hMSCs proliferation and glycolysis but reduce their mitochondrial metabolism with hypoxia amplifying these effects. CONCLUSIONS: Cells that switch to glycolysis during co-cultivation, particularly under hypoxia, benefit most from these low oxygen conditions. Due to this leukemic cells' response heterogeneity, targeting microenvironmental interactions and oxygen levels is crucial for personalized leukemia therapy. Advancing co-cultivation models, particularly through innovations like spheroids, can further enhance in vitro studies of primary leukemic cells and support the testing of novel therapies.

• Klíčová slova
• Co-cultivation, Feeders, Hypoxia, Leukemic cells, Tumor microenvironment,
• MeSH
• fibroblasty * metabolismus   MeSH
• glykolýza MeSH
• hypoxie buňky MeSH
• kokultivační techniky metody   MeSH
• leukemie * patologie   metabolismus   MeSH
• lidé MeSH
• mezenchymální kmenové buňky * metabolismus   MeSH
• mitochondrie metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• proliferace buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH