Epithelial-mesenchymal transition (EMT) is a crucial process with significance in the metastasis of malignant tumors. It is through the acquisition of plasticity that cancer cells become more mobile and gain the ability to metastasize to other tissues. The mesenchymal-epithelial transition (MET) is the return to an epithelial state, which allows for the formation of secondary tumors. Both processes, EMT and MET, are regulated by different pathways and different mediators, which affects the sophistication of the overall tumorigenesis process. Not insignificant are also cancer stem cells and their participation in the angiogenesis, which occur very intensively within tumors. Difficulties in effectively treating cancer are primarily dependent on the potential of cancer cells to rapidly expand and occupy secondarily vital organs. Due to the ability of these cells to spread, the concept of the circulating tumor cell (CTC) has emerged. Interestingly, CTCs exhibit molecular diversity and stem-like and mesenchymal features, even when derived from primary tumor tissue from a single patient. While EMT is necessary for metastasis, MET is required for CTCs to establish a secondary site. A thorough understanding of the processes that govern the balance between EMT and MET in malignancy is crucial.

• Klíčová slova
• Cancer stem Cells, Circulating Tumor cell (CTC), Epithelial-mesenchymal Transition (EMT), Mesenchymal-epithelial Transition (MET),
• MeSH
• epitelo-mezenchymální tranzice * MeSH
• fenotyp MeSH
• kmenové buňky metabolismus   cytologie   patologie   MeSH
• lidé MeSH
• nádorové cirkulující buňky * patologie   metabolismus   MeSH
• nádorové kmenové buňky * patologie   metabolismus   MeSH
• nádory patologie   metabolismus   MeSH
• patologická angiogeneze * patologie   MeSH
• proliferace buněk genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

In vitro meat production presents a potential viable alternative for meat consumption, which could provide the consumer with a product indistinguishable from the original, with very similar nutritional and culinary values. Indeed, the alternative products currently accessible often lack comparable nutritional value or culinary attributes to their animal-derived counterparts. This creates challenges for their global acceptance, particularly in countries where meat consumption holds cultural significance. However, while cultured meat research has been progressing rapidly in recent years, some significant obstacles still need to be overcome before its possible commercialization. Hence, this review summarizes the most current knowledge regarding the history of cultured meat, the currently used cell sources and methods used for the purpose of in vitro meat production, with particular focus on the role of bioreactors, scaffolds and microcarriers in overcoming the current obstacles. The authors put the potential microcarrier and scaffold-based solutions in a context, discussing the ways in which they can impact the way forward for the technology, including the use of considering the potential practical and societal barriers to implementing it as a viable food source worldwide.

• Klíčová slova
• bioreactor, cultured meat, in vitro culture, microcarriers, scaffold,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Genes influencing oocyte maturation may be valuable for predicting their developmental potential, as well as discerning the mechanistic pathways regulating oocyte development. In the presented research microarray gene expression analysis of immature and in vitro matured porcine oocytes was performed. Two groups of oocytes were compared in the study: before (3 × n = 50) and after in vitro maturation (3 × n = 50). The selection of viable oocytes was performed using the brilliant cresyl blue (BCB) test. Furthermore, microarrays and RT-qPCR was used to analyze the transcriptome of the oocytes before and after IVM. The study focused on the genes undergoing differential expression in two gene-ontology groups: "Cellular response to hormone stimulus" and "Cellular response to unfolded protein", which contain genes that may directly or indirectly be involved in signal transduction during oocyte maturation. Examination of all the genes of interest showed a lower level of their expression after IVM. From the total number of genes in these gene ontologies ten of the highest change in expression were identified: FOS, ID2, BTG2, CYR61, ESR1, AR, TACR3, CCND2, EGR2 and TGFBR3. The successful maturation of the oocytes was additionally confirmed with the use of lipid droplet assay. The genes were briefly described and related to the literature sources, to investigate their potential roles in the process of oocyte maturation. The results of the study may serve as a basic molecular reference for further research aimed at improving the methods of oocyte in vitro maturation, which plays an important role in the procedures of assisted reproduction.

• Klíčová slova
• Microarray, Mitochondrial activity, Oocyte maturation, Pig,
• MeSH
• eosin chemie   MeSH
• hematoxylin chemie   MeSH
• hormony genetika   metabolismus   MeSH
• IVM techniky * MeSH
• kultivované buňky MeSH
• lipidy analýza   MeSH
• oocyty růst a vývoj   metabolismus   MeSH
• oxaziny chemie   MeSH
• prasata MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Brilliant Cresyl Blue MeSH Prohlížeč
• eosin MeSH
• hematoxylin MeSH
• hormony MeSH
• lipidy MeSH
• oxaziny MeSH

The primary function of ovarian granulosa cells (GCs) is the support of oocytes during maturation and development. Molecular analyses of granulosa cell-associated processes, leading to improvement of understanding of the cell cycle events during the formation of ovarian follicles (folliculogenesis), may be key to improve the in vitro fertilization procedures. Primary in vitro culture of porcine GCs was employed to examine the changes in the transcriptomic profile of genes belonging to "cell cycle", "cell division", "cell cycle process", "cell cycle phase transition", "cell cycle G1/S phase transition", "cell cycle G2/M phase transition" and "cell cycle checkpoint" ontology groups. During the analysis, microarrays were employed to study the transcriptome of GCs, analyzing the total RNA of cells from specific periods of in vitro cultures. This research was based on material obtained from 40 landrace gilts of similar weight, age and the same living conditions. RNA was isolated at specific timeframes: before the culture was established (0 h) and after 48 h, 96 h and 144 h in vitro. Out of 133 differentially expressed genes, we chose the 10 most up-regulated (SFRP2, PDPN, PDE3A, FGFR2, PLK2, THBS1, ETS1, LIF, ANXA1, TGFB1) and the 10 most downregulated (IGF1, NCAPD2, CABLES1, H1FOO, NEK2, PPAT, TXNIP, NUP210, RGS2 and CCNE2). Some of these genes known to play key roles in the regulation of correct cell cycle passage (up-regulated SFRP2, PDE3A, PLK2, LIF and down-regulated CCNE2, TXNIP, NEK2). The data obtained provide a potential reference for studies on the process of mammalian folliculogenesis, as well as suggests possible new genetic markers for cell cycle progress in in vitro cultured porcine granulosa cells.

• Klíčová slova
• Granulosa cells, Microarray, Ovarian follicle, Pig, Primary culture,
• MeSH
• buněčný cyklus genetika   MeSH
• folikulární buňky cytologie   MeSH
• kultivované buňky MeSH
• ovariální folikul cytologie   MeSH
• prasata MeSH
• stanovení celkové genové exprese MeSH
• transkriptom * MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx's behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.

• MeSH
• akvaporiny genetika   metabolismus   MeSH
• biologické modely * MeSH
• buněčné kultury * MeSH
• endometrium cytologie   MeSH
• epitelové buňky metabolismus   MeSH
• konexiny genetika   metabolismus   MeSH
• kultivované buňky MeSH
• prasata MeSH
• proliferace buněk genetika   MeSH
• regulace genové exprese * MeSH
• tvar buňky MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akvaporiny MeSH
• konexiny MeSH

Proper course of folliculogenesis and oogenesis have an enormous impact on female fertility. Both processes take place in the ovary and involve not only the maturing germ cell, but also few types of somatic cells that assist the ovarian processes and mediate the dialog with the oocyte. These cells, granulosa and theca, are heavily involved in essential reproductive processes, such as ovulation, fertilization, and embryo implantation. In this study, we have used the expressive microarray approach to analyze the transcriptome of porcine granulosa cells, during short-term in vitro culture. We have further selected differentially expressed gene ontologies, involved in cell proliferation, migration, adhesion, and tissue development, namely, "cell-cell adhesion," "cell motility," "cell proliferation," "tissue development," and "tissue migration" to screen them for the possibility of discovery of new markers of those processes. A total of 303 genes, expression of which varied significantly in different culture periods and belonged to the analyzed ontology groups, were detected, of which 15 that varied the most (between 0 and 48 h of culture) were selected for validation. As the validation confirmed the transcriptomic patterns, 10 genes of biggest changes in expression (CAV1, IGFBP5, ITGB3, FN1, ITGA2, LAMB1, POSTN, FAM83D, KIF14, and CDK1) were analyzed, described, and referred to the context of the study, with the most promising new markers and further proof for the viability of the currently recognized ones detailed. Overall, the study provided valuable insight into the molecular functioning of in vitro granulosa cell cultures.

• Klíčová slova
• Maturation (IVM), adhesion, cellular migration, porcine granulosa cells, proliferation,
• MeSH
• biologické markery metabolismus   MeSH
• buněčná adheze genetika   MeSH
• folikulární buňky cytologie   metabolismus   fyziologie   MeSH
• kultivované buňky MeSH
• pohyb buněk genetika   MeSH
• prasata MeSH
• proliferace buněk genetika   MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• stanovení celkové genové exprese MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH

The human ovarian granulosa cells (GCs) surround the oocyte and form the proper architecture of the ovarian follicle. The ability of GCs to proliferate and differentiate in the conditions of in vitro culture has been proven. However, there is still a large field for extensive investigation of molecular basics, as well as marker genes, responsible for these processes. This study aimed to find the new marker genes, encoding proteins that regulate human GCs in vitro capability for proliferation and differentiation during long-term primary culture. The human follicular GCs were collected from hyper-stimulated ovarian follicles during IVF procedures and transferred to a long-term in vitro culture. The culture lasted for 30 days, with RNA samples isolated at days 1, 7, 15, 30. Transcriptomic analysis was then performed with the use of Affymetrix microarray. Obtained results were then subjected to bioinformatical evaluation and sorting. After subjecting the datasets to KEGG analysis, three differentially expressed ontology groups "cell differentiation" (GO:0030154), "cell proliferation" (GO:0008283) and "cell-cell junction organization" (GO:0045216) were chosen for further investigation. All three of those ontology groups are involved in human GCs' in vitro lifespan, proliferation potential, and survival capability. Changes in expression of genes of interest belonging to the chosen GOs were validated with the use of RT-qPCR. In this manuscript, we suggest that VCL, PARVA, FZD2, NCS1, and COL5A1 may be recognized as new markers of GC in vitro differentiation, while KAT2B may be a new marker of their proliferation. Additionally, SKI, GLI2, FERMT2, and CDH2 could also be involved in GC in vitro proliferation and differentiation processes. We demonstrated that, in long-term in vitro culture, GCs exhibit markers that suggest their ability to differentiate into different cells types. Therefore, the higher expression profile of these genes may also be associated with the induction of cellular differentiation processes that take place beyond the long-term primary in vitro culture.

• Klíčová slova
• Differentiation, Granulosa cells, Microarrays, Proliferation, Stem cells,
• MeSH
• adhezní spoje metabolismus   MeSH
• buněčná adheze genetika   MeSH
• buněčná diferenciace genetika   MeSH
• dospělí MeSH
• folikulární buňky cytologie   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• ovarium cytologie   MeSH
• proliferace buněk genetika   MeSH
• upregulace * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The physiological processes that drive the development of ovarian follicle, as well as the process of oogenesis, are quite well known. Granulosa cells are major players in this occurrence, being the somatic element of the female gamete development. They participate directly in the processes of oogenesis, building the cumulus-oocyte complex surrounding the ovum. In addition to that, they have a further impact on the reproductive processes, being a place of steroid sex hormone synthesis and secretion. It is known that the follicle development creates a major need for angiogenesis and blood vessel development in the ovary. In this study, we use novel molecular approaches to analyze markers of these processes in porcine granulosa cultured primarily in vitro. The cells were recovered from mature sus scrofa specimen after slaughter. They were then subjected to enzymatic digestion and culture primarily for a short term. The RNA was extracted from cultures in specific time periods (0h, 24h, 48h, 96h, and 144h) and analyzed using expression microarrays. The genes that exhibited fold change bigger than |2|, and adjusted p-value lower than 0.05, were considered differentially expressed. From these, we have chosen the members of "angiogenesis," "blood vessel development," "blood vessel morphogenesis," "cardiovascular system development," and "vasculature development" for further selection. CCL2, FGFR2, SFRP2, PDPN, DCN, CAV1, CHI3L1, ITGB3, FN1, and LOX which are upregulated, as well as CXCL10, NEBL, IHH, TGFBR3, SCUBE1, IGF1, EDNRA, RHOB, PPARD, and SLITRK5 genes whose expression is downregulated through the time of culture, were chosen as the potential markers, as their expression varied the most during the time of culture. The fold changes were further validated with RT-qPCR. The genes were described, with special attention to their possible function in GCs during culture. The results broaden the general knowledge about GC's in vitro molecular processes and might serve as a point of reference for further in vivo and clinical studies.

• MeSH
• cévy růst a vývoj   metabolismus   MeSH
• folikulární buňky cytologie   metabolismus   MeSH
• fyziologická neovaskularizace genetika   MeSH
• lidé MeSH
• morfogeneze genetika   MeSH
• oocyty růst a vývoj   MeSH
• oogeneze genetika   MeSH
• ovariální folikul růst a vývoj   MeSH
• ovarium růst a vývoj   metabolismus   MeSH
• prasata MeSH
• primární buněčná kultura MeSH
• proteosyntéza genetika   MeSH
• vývojová regulace genové exprese genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The similarity between humans and pigs, when it comes to tissue morphology, makes Sus scrofa not only a good research model, but also a potential source of cells for tissue engineering. Cell samples obtained from the pig donor, could be influenced in vitro, in order to become a source of tissue material for xenotransplantation, reconstructive and regenerative medicine. Significant amounts of data point to especially major similarities in pig and human reproductive systems. Because of that, particular scientific focus is centered on research concerning porcine COCs, theca and granulosa cells in primary cultures. One of the aspects of the reproductive process, that is still largely undiscovered, is the interaction between preimplantation blastocyst and maternal uterine tissues. In this study, we used molecular analysis techniques, such as RT-qPCR and immunocytochemistry, to analyze the expression and distribution of cytokeratin 18 and panCytokeratins 8, 18 and 19 and vimentin in porcine luminal endometrial epithelial cells, coupled with analysis of their behavior in RTCA. The results have confirmed the presence of epithelial, as well as stromal cell markers in the cells, varying in levels at different stages of culture. They have also given insight into the modes of proliferation and differentiation of studied cells in in vitro culture, as well as providing additional proof for the possible mesenchymal transdifferentiation of epithelial cells.

• MeSH
• biologické markery metabolismus   MeSH
• biologické modely MeSH
• buněčná diferenciace MeSH
• buňky stromatu cytologie   metabolismus   MeSH
• časové faktory MeSH
• endometrium cytologie   MeSH
• epitelové buňky cytologie   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• modely u zvířat MeSH
• prasata MeSH
• proliferace buněk * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH

Extraordinary abilities for continuous proliferation and differentiation, associated with constant renewal triggered by stimulation from the mastication process, together with the relative lack of aesthetic complications associated with post-surgery healing, have highlighted buccal pouch mucosa as a potential source of explants that could be used in transplantation and tissue engineering. Additionally, this tissue plays a major role in the oral drug delivery process, which brings special interest to its molecular properties in the context of new drug development. There is therefore a need to analyse the exact mechanisms of oral mucosa functioning, especially when it comes to the processes that are associated with the potential clinical applications. In this study we analysed a complete transcriptome of long-term in vitro cultures of porcine buccal pouch oral mucosa cells. Using a microarray approach, we focused on genes associated with cellular metabolic processes, signalling and adhesion, from 4 gene ontology groups: "Positive regulation of cellular component movement", "Positive regulation of cellular process", "Positive regulation of intracellular signal transduction" and "Single organism cell adhesion". Nineteen genes (CCL8, CXCL2, PLK2, DUSP5, PTGS2, LIF, CCL2, ATP1B1, REL, ITGB3, SCARB1, UGCG, PDPN, LYN, ETS1, FCER1G, TGFB1, RFC4, LMO2) with fold changes higher than |2| and p value Extraordinary abilities for continuous proliferation and differentiation, associated with constant renewal triggered by stimulation from the mastication process, together with the relative lack of aesthetic complications associated with post-surgery healing, have highlighted buccal pouch mucosa as a potential source of explants that could be used in transplantation and tissue engineering. Additionally, this tissue plays a major role in the oral drug delivery process, which brings special interest to its molecular properties in the context of new drug development. There is therefore a need to analyse the exact mechanisms of oral mucosa functioning, especially when it comes to the processes that are associated with the potential clinical applications. In this study we analysed a complete transcriptome of long-term in vitro cultures of porcine buccal pouch oral mucosa cells. Using a microarray approach, we focused on genes associated with cellular metabolic processes, signalling and adhesion, from 4 gene ontology groups: "Positive regulation of cellular component movement", "Positive regulation of cellular process", "Positive regulation of intracellular signal transduction" and "Single organism cell adhesion". Nineteen genes (CCL8, CXCL2, PLK2, DUSP5, PTGS2, LIF, CCL2, ATP1B1, REL, ITGB3, SCARB1, UGCG, PDPN, LYN, ETS1, FCER1G, TGFB1, RFC4, LMO2) with fold changes higher than |2| and p value less than 0.05 were identified, described in context and analysed. While the study needs much further validation to become applicable in a clinical environment, it yields valuable information about the transcriptomic basis of oral mucosal cell functioning in vitro, that might serve as a reference for further research, aiming to apply this knowledge in clinical situations.0.05 were identified, described in context and analysed. While the study needs much further validation to become applicable in a clinical environment, it yields valuable information about the transcriptomic basis of oral mucosal cell functioning in vitro, that might serve as a reference for further research, aiming to apply this knowledge in clinical situations.

• MeSH
• buněčná adheze genetika   MeSH
• buněčné kultury MeSH
• genetické markery genetika   MeSH
• kultivované buňky MeSH
• prasata * MeSH
• signální transdukce genetika   MeSH
• stanovení celkové genové exprese * MeSH
• tvář MeSH
• ústní sliznice cytologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• genetické markery MeSH