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Therapeutic potential of Sertoli cells for the treatment of neurodegenerative diseases
Djekić, Danijela ; Krylov, Vladimír (advisor) ; Fikarová, Natálie (referee)
Neurodegenerative diseases are widespread and their incidence is increasing worldwide. They become the main cause of incapacity for work and affect not only their carriers, but also their immediate surroundings. One of the causes of the development of these diseases is the inflammatory process. Sertoli cells are capable of locally and systemically inducing an anti- inflammatory response of the immune system, which has recently been used for allotransplantation of grafts of various origins. It is the anti-inflammatory nature of these cells that can be interesting for the treatment of neurodegenerative diseases.
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Comparison of immunomodulatory properties of mesenchymal stem cells and Sertoli cells
Porubská, Bianka ; Krulová, Magdaléna (advisor) ; Komrsková, Kateřina (referee) ; Filipp, Dominik (referee)
Cell therapies are increasingly considered in preclinical studies and in the future of medicine. The main cell type investigated in this manner is mesenchymal stem cells (MSCs), because of their strong immunomodulatory properties. The efficacy of the therapy depends on various aspects, such as the viability and source of MSCs, the purity of the cell suspension and many more. There is a need for more tailored therapy and the use of cell type better fitting for the specific pathology. Sertoli cells (SCs) are deemed by some authors to be a kind of MSCs, namely because of their similar immunomodulatory properties. Because they reside in the seminiferous tubules in the testes, they are a promising candidate for the treatment of inflammatory pathologies of testicular tissue, such as bacterial infection-induced infertility. In vitro comparison of the ability of MSCs and SCs to differentiate into mesenchymal cell lineages such as osteocyte, chondrocyte, and adipocyte showed success in the case of SCs, providing evidence for their mesenchymal origin. The effect of MSCs or SCs on activated immune cells in vitro showed immunosuppression in both cases with distinct features. MSCs suppressed Th17 cell activation and IL-17 production by CD4+ T cells and SCs down-regulated TNFα and IL-2 production by these cells,...
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Therapeutic influence of Sertoli cells on the effects of oxidative stress in germ cells
Hynková, Marie ; Tlapáková, Tereza (advisor) ; Frolíková, Michaela (referee)
Sertoli cells are somatic cells found in the testes and have a function in supporting spermatogenesis and germ cell nutrition, but also in maintaining an anti-inflammatory environment and forming the blood-testicular barrier in the testes. They are intensively researched for their immunomodulating properties which can be used in transplant medicine or as a therapeutic tool in regenerative medicine. Following previous research confirming that mouse Sertoli cells share some characteristics of mesenchymal stem cells and have multipotent differentiation potential in vitro, experiments were conducted to determine their therapeutic capabilities. Sertoli cells have demonstrated the ability to promote the maintenance of sperm motility during testicular inflammation and reduce the amount of immotile sperm in vivo. Based on preliminary results, they can also reduce the amount of ROS produced by testicular cells affected by oxidative stress in vitro. Keywords: Sertoli cells, germ cells, infertility, oxidative stress, ROS
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Effect of estrogens on in vitro models of testicular tissue and spermatogenesis
Jursová, Pavlína ; Děd, Lukáš (advisor) ; Tlapáková, Tereza (referee)
Although estrogens are primarily known for their functions in female reproductive system, their effect on male reproductive functions has also been well established. Physiological estrogen concentration is essential for a proper spermatogenesis. Estrogens regulate many functions in testicular tissue, including proliferation and apoptosis of all testicular cell types, dynamic restructuring of cell-cell junctions in the testis, and post-translation modifications of histones. Hence, the aim of this thesis was to study effect of estrogens on in vitro models of testicular tissue and spermatogenesis and thus to address their functions in testicular tissue more deeply. This project includes testicular organoid cultivation for further usage as in vitro model of spermatogenesis. To addresss the effect of various avaliable estrogen forms, experiments on MCF-7 cell line were done. Finally experiments with in vitro model of testicullar tissue - TM4 Sertoli cell line were done. In order to fulfill the aims and verify the hypotheses, some advanced methods such as CLARITY volume confocal imaging and holographic microscopy were used. It was found that estrogens can affect Sertoli cell morphology and the expression of some genes involved in cell-cell junction dynamics. Furthermore the process of spermatogenesis was...
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Mitochondrial transfer-mediated modulatory action of stem cells on immune cells
Somova, Veronika ; Krulová, Magdaléna (advisor) ; Balounová, Jana (referee)
Stem cells use different mechanisms of intercellular communication to modulate an immune response. Mitochondrial transfer is one of the mechanisms which induce metabolic changes, support cell survival, and change the phenotype of immune cells. Nevertheless, little is known about the mechanism used for transfer of mitochondria between different cell populations and the faith of mitochondria inside the acceptor cell. This thesis aims to describe the mechanism of transfer and the provided modulation. Factors that could affect mitochondrial transfer including reactive oxygen species production, apoptosis and mitochondria function were analyzed. And the impact of mitochondrial transfer on cell survival and mitophagy was described. The next aim was to compare the ability of mesenchymal stem cells (MSC) and Sertoli cells (SC) to transfer mitochondria, with MSC being more productive in the transfer of mitochondria than SC. Significant differences in the presence of mitochondria from donor MSC or SC in individual populations of immune cells were also detected. To explain these findings, the impact of reactive oxygen species on the transfer of mitochondria was analyzed in detail, although it wasn't confirmed. However, it needs to be highlighted that mitophagy plays an important role before and after...
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The role of Fbxo38 ubiquitin ligase in mouse spermatogenesis
Zobalová, Eliška ; Čermák, Lukáš (advisor) ; Stopka, Pavel (referee)
Cullin-dependent ubiquitin ligases are responsible for the regulation of most cellular processes. Despite their mutated forms being the cause of many human diseases, their physiological roles are not sufficiently described. In the presented results, we focused on the physiological role of ubiquitin ligase SCFFBXO38 (SKP1-CULLIN1-FBXO38), whose mutated forms are responsible for the progression of distal neuropathy. Preparation of mouse model deficient in FBXO38 revealed that homozygous pups were born in a lower than expected ratio. Animals were growth-retarded, both at the level of the whole organism and individual organs, especially the liver and testes. Males with a deletion in the Fbxo38 gene had significantly lower reproductive capacity, which was associated with lower production of mature sperm and pathological changes in the structure of seminiferous tubules. We found that the FBXO38 protein is functionally expressed in Sertoli cells responsible for regulating spermatogenesis and seminiferous tubules integrity. Detailed analysis of spermatogenic populations revealed a defect at the level of spermatocyte differentiation. The dynamics of this differentiation depend on the hematotesticular barrier functional integrity formed by the intercellular junctions of Sertoli cells. We confirmed that the...
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Differentiation of Sertoli cell progenitors and preparation of testicular 3D cultures of Xenopus tropicalis.
Slováková, Lucie ; Tlapáková, Tereza (advisor) ; Hovořáková, Mária (referee)
Sertoli cells represent the only somatic cell type within the seminiferous tubules with direct contact to germ cells. Sertoli cells significantly contribute to the development of the testicular niche in a male embryo. Their role during postnatal life is in the regulation and nutrition of germ cells and the formation of the blood-testis barrier to protect these cells. In our laboratory, we have been successful in establishing a cell line of X. tropicalis immature Sertoli cells (XtiSCs) derived from juvenile testes of X. tropicalis. The objective of this thesis was to induce the differentiation process of XtiSCs into mature Sertoli cells. In vitro experiments using several factors or primary culture from adult male X. tropicalis did not show any mature markers in differentiated XtiSCs. Another experiment using cell culture derived from pubertal mice was partially successful in the induction of the differentiation process. These results indicate that XtiSCs do have some differentiation potential into mature Sertoli cells. Part of this work was to test the ability of testicular cells isolated from juvenile males of X. tropicalis to form de novo organoids. In vitro experiments were successful when these cells were cultured in a three-layer matrigel.
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Study of regenerative potential of Sertoli cell progenitors in Xenopus tropicalis tadpoles after tail amputation.
Wróblová, Aneta ; Krylov, Vladimír (advisor) ; Procházka, Jan (referee)
African clawed frogs (Xenopus) represent an ideal model organism for study of regeneration mechanisms. In frogs, complete regeneration occurs in the tadpole stage. In later stages the regeneration capacity is lost. The Laboratory of Developmental biology was successful in establishment of cell culture called Xenopus tropicalis immature Sertoli cells (XtiSCs) derived from X. tropicalis testes. These cells are common progenitors of Sertoli cells and peritubular myoid cells. XtiSCs show similar characteristics as mesenchymal stem cells. MSCs hold interest of scientists for their immunomodulatory properties and multipotent differential and regeneration potential. In this thesis, we studied regeneration and migration potential of XtiSCs after X. tropicalis tadpole's tail amputation in developmental stage 47 - 50. Transgenic XtiSCs culture expressing RFP was prepared to facilitate transplantation experiments. Transplantation experiments showed preferential migration of XtiSCs into the site of injury. XtiSCs transplantations in X. laevis tadpoles with downregulated NO synthases eNOS and nNOS revealed their migratory dependence on nitric oxide signalization. Imunocytochemical staining of XtiSCs in vitro showed positive iNOS, nNOS and Pax7 expression. Imunohistochemical staining of tadpole's tail vibratome...
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Regenerative potential of Sertoli cell progenitors regarding heart injury in Xenopus tropicalis
Onhajzer, Jakub ; Krylov, Vladimír (advisor) ; Procházka, Jan (referee)
Cardiac failure is one of the leading cause of deaths worldwide. Potential therapeutic approach, which overcome invasive organ transplantation and delivery of immunosuppressive drugs, is lacking nowadays. However, research of mesenchymal stem cells (MSCs) therapy displays immunomodulation potential, which can further promote variety of organ regeneration without need of drug treatment. Xenopus tropicalis immature Sertoli cells (XtiSCs) culture was established in our laboratory from juvenile Xenopus tropicalis male. XtiSCs possess immunomodulatory capacity and differentiation to cardiomyocytes after the treatment with the inhibitor of glycogen synthase kinase-3 (GSK-3) CHIR99021. To test the survival rate of transplanted XtiSCs we firstly microinjected treated cells directly inside tadpole's heart. XtiSCs proliferated there for the whole tested time period (30 days). However, after direct heart XtiSCs injection and subsequent cardiac injury in adult frog, no cells were localized in wound area. Thus, we focused on remote control of cardiac regeneration using XtiSCs without CHIR99021 treatment. We injected cells inside skeletal muscle bed and confirmed their survival and proliferation. Moreover, if cells were transplanted 3 days before heart injury, it resulted in significant reduction of fibronectin...
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Study of mechanisms of Sertoli cell survival in xenogeneic organism
Porubská, Bianka ; Krulová, Magdaléna (advisor) ; Anděra, Ladislav (referee)
Sertoli cells (SCs) are somatic cells located in the testes. They are the only cells in direct contact with germ cells and play a key role in process of spermatogenesis. New insights in the biology of SCs are highlighting the immunological function of these cells: germ cells protection by maintaining the immunoprotective niche, creating the blood- testis barrier and local modulation of the immune response to spermatic cells. Immunomodulatory activity of SCs is preserved after their allo- and xenogeneic transplantation, and thus SCs prolongs survival not only of themselves but also of cells transplanted with them. The aim of this thesis was to study the survival and migration of SCs precursors (TSC) in mice recipients. The project is employing the neonatal tolerance phenomenon and evolutionary distinct donor organism, Xenopus tropicalis, to monitor conserved mechanisms of immune system (IS) modulation using SCs. SCs were detectable in the lungs and thymus 7 days after transplantation. The phenotype of immune cells was not altered 30 days after transplantation, however we detected changes in cytokine environment, namely increased levels of cytokines typical for Th2 and Treg immune responses. In vitro experiments further confirmed IS modulation by SCs - changing the phenotype of macrophages to alternatively...
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