|
|
The effect of environmental factors on mesenchymal stem cells
Hlávka, Jakub ; Krulová, Magdaléna (advisor) ; Vištejnová, Lucie (referee)
Mesenchymal stem cells (MSCs) have the potential to be a valuable therapeutic tool due to their capacity for differentiation and immunomodulation. However, in order to ensure their efficacy in clinical applications, it is necessary to gain a comprehensive understanding of the factors influecing their properties. This bachelor thesis investigates the influence of environmental factors, including physical activity, temperature and light exposure, on MSCs functionality. Additionally it desribes the underlying mechanisms involved in the action of these factors. Finally, it evaluates the theoretical implications of exploiting environmental cues to enhance MSCs properites. An understanding of how environmental conditions shape MSCs behavior is essential for optimising their therapeutic potential and advancing regenerative medicine strategies. Key words: mesenchymal stem cells, environmental factors, physical activity, temperature, light
|
|
|
Impact of nanomaterials on mesenchymal stem cells and tissue regeneration
Echalar, Barbora ; Holáň, Vladimír (advisor) ; Hubálek Kalbáčová, Marie (referee) ; Buchtová, Marcela (referee)
Nanomaterials (NMs) are widely used in medicine for their antimicrobial properties. They are part of antibacterial coatings, creams, pharmaceutical vehicles or additives in drugs and other medical products. However, the impact of NMs on human organism is still not completely established. Nanoparticles (NPs) penetrate the cell membrane and enter to intracellular compartments including the nuclei. Different types of NPs could have various side effects on cell functions. These side effects include the damage of stem cells (SCs) or immune cells lead to slower regeneration and impaired wound healing. Therefore, the simultaneous application of NPs during SC therapy could decrease the therapeutic abilities of SCs. One type of SCs tested in clinical therapies nowadays are mesenchymal stem cells (MSCs). Therefore, we studied the impact of metal NPs (i.e. silver, copper oxide, zinc oxide and titanium dioxide) on characteristics and functional properties of mouse MSCs. Additionally, the effect of NPs on the expression of phenotypic markers, metabolic activity, differentiation potential, expression of genes for immunoregulatory molecules and on production of cytokines and growth factors was analyzed. We found that all types of tested NPs had a negative impact on the activity of MSCs and thus could alter tissue...
|
|
|
Modulation of mitochondrial transfer by influencing mesenchymal stem cells
Fráňová, Markéta ; Krulová, Magdaléna (advisor) ; Rohlenová, Kateřina (referee)
Mesenchymal stem cells (MSCs) have the ability to modulate the immune response. They use several mechanisms to affect the function of immune cells, and mitochondrial transfer is one of them. Recieving mitochondria from MSCs induces metabolic changes in immune cells, thereby promoting their shift to an anti-inflammatory phenotype. Due to their properties, MSCs have a potencial to be used in therapies, for example in a treatment of autoimmune diseases. The problem of MSCs-based therapies is their low efficacy, mainly due to the high mortality of stem cells after transplantation. In order to achieve at least some effect, the large number of cells is needed for application. The required number of cells can be obtained only by in vitro expansion. However, a long-term culture has a negative impact on MSCs and their immunomodulatory properties. Enhancing MSCs function could increase the efficacy of MSCs-based therapies. The aim of this thesis was to determine whether mitochondrial transfer can be modulated by stimulation of MSCs with selected factors. MSCs were treated with rapamycin, insulin-like growth factor 1 (IGF-1), interferon gamma, or oligomycin. Then the effect of these factors on mitochondria and their transfer to immune cells, metabolism, and immunomodulatory properties of MSCs was analyzed. We...
|
|
|
Fluorescent Methods in Research of Eukaryotic Cells
Chmelíková, Larisa ; Babula, Petr (referee) ; Pešl,, Martin (referee) ; Provazník, Ivo (advisor)
Tato práce zkoumá aplikaci fluorescenčních metod používaných v in vitro studiích v oblasti regenerace srdeční tkáně. Konfokální fluorescenční mikroskopie je vhodnou mikroskopickou technikou pro výzkum v této oblasti, protože umožňuje vizualizaci 3D struktur a distribuce buněk ve 3D modelech. Používané fluorescenční markery by měly být dlouhodobě stabilní, biokompatibilní a netoxické pro živé buňky. V současné době je použití nanočástic jako superparamagnetické nanočástice oxidu železa (SPION) velmi populární; velké množství studií ukazuje, že jsou vhodné pro dlouhodobé experimenty. Tento výzkum využívá superparamagnetické maghemitové nanočástice svázaným rhodaminem na jejich povrchu (SAMN-R) a popisuje jejích excitační a emisní spektrum, velikost a lokalizaci vbuňkách. Stanovení toxicity bylo provedeno měřením reaktivních forem kyslíku (ROS) a nekvantitativním měřením pomocí fluorescenční mikroskopie bylo zjištěno, že hodnota dávky 20 µg·cm-2 je optimální pro aplikaci na živé buňky. Dále byl zkoumán vliv aplikace SAMN-R na buněčnou proliferaci a motilitu, kdy ve studii buněčné proliferace a scratch assay byla použita buněčná linie fibroblastů 3T3. Poté byla studována migrace jednotlivých buněk s použitím mezenchymálních kmenových buněk (MSCs), izolovaných zlidské tukové tkáně. Následná statistická analýza nepotvrdila, že by aplikace SAMN-R měla významný vliv na buněčnou proliferaci, kolektivní migraci nebo na migraci jednotlivých buněk. Lze tedy předpokládat, že SAMN-R jsou vhodným fluorescenčním markerem pro výzkum živých buněk, včetně experimentů voblasti regenerace tkáně. MSC buňky izolované z tukové tkáně mají velký potenciál v regeneraci srdeční tkáně. Jejich interakce s buněčnou linií srdečních svalových buněk HL-1 byly studovány pomocí scratch assay, kdy se tento model jeví jako nadějný a vhodný pro studium buněčných kontaktů a jejich roli přiregeneraci buněk.
|
|
| |
|
|
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,...
|
|
|
Human Dental Pulp Stem Cells Cultured in Xenogeneic-Free Supplemented Media
Suchánková Kleplová, Tereza ; Jouklová, Nela (advisor) ; Merglová, Vlasta (referee) ; Bartoňová, Marie (referee)
Human dental pulp stem cells cultured in xenogeneic-free supplemented media Summary Introduction: The topic of the study is the cultivation of dental pulp stem cells (hDPSCs) in a xenogeneic-free culture medium. It is not permissible to use cells upon growing under the influence of xenogeneic (extraneous) substances in human clinical practice. The most frequently used in cultivation of hDPSCs is fetal calf serum (FCS/FBS). Unfortunately, these supplements are widespread in hMSCs cultivation, and all gold standard hMSCs properties were postulated in cells cultivated using these supplements. This raises the basic question if and how xenogeneic blood derivatives affect the properties of cells and their growth characteristics. There are two options for replacing these xenogeneic substances in the culture medium: the so-called serum-free media, or human blood supplements, ideally autologous ones. The conducted research was aimed at identifying the effects of xenogeneic and human blood supplements on basic hDPSCs characteristics that are fundamental to introduce the cell therapy into regular medical practice. Method: By culturing 12 hDPSC lines obtained from adult, deciduous, and natal teeth in 12 different culture media, we investigated the effect of FCS, human blood derivatives, i.e., blood plasma (HP), and...
|
|
|
Mitochondrial transfer and its role in regenerative properties of mesenchymal stem cells
Jaborová, Natálie ; Krulová, Magdaléna (advisor) ; Nahácka, Zuzana (referee)
Mitochondrial transfer represents a form of intercellular communication. In this process, mitochondria are delivered from the donor cell to the recipient cell through several structures. The transfer of mitochondria is observed under pathological and physiological conditions and is accompanied by specific signaling. Mitochondria uptake by injured cells promotes tissue regeneration. This bachelor thesis discusses general knowledge of mitochondrial transfer focusing on mesenchymal stem cells (MSCs) as a donor cell type. Furthermore, the regenerative effect of MSC-derived mitochondria transfer to cells damaged by various pathologies is summarized. In this regard, MSCs have significant therapeutic potential that could be used in future clinical strategies for a number of diseases. Keywords: mitochondria, transfer, mesenchymal stem cells, regeneration
|
|
|
Immunomodulatory and differentiation properties of MSCs in a mouse model of the injured cornea and retina
Kössl, Jan ; Holáň, Vladimír (advisor) ; Vištejnová, Lucie (referee) ; Heissigerová, Jarmila (referee)
Stem cells, in general, represent the potential for treating many diseases and disorders that are currently difficult to treat or the therapy has many side effects. One of the stem cells widely investigated these days are mesenchymal stem cells (MSCs). MSCs have the considerable immunomodulatory and regenerative potential for treating degenerative disorders and severe damage to various parts of the eye or other organs. Likewise, their application could serve as supportive therapy in corneal transplantation and other eye inflammatory conditions. In this study of immunomodulatory properties of MSCs, we have focused mainly on their ability to differentiate into cells of different tissue types (in our case, corneal epithelium and retina), their production of immunomodulatory molecules in the inflammatory environment, their ability to migrate to the site of the injury, and their local anti-inflammatory, regenerative, and anti-apoptotic effects. In addition, we tested the therapeutic effects of MSCs in a mouse model of ocular surface injury and a model of retinal degeneration. Finally, we investigated the mechanism of this effect in in vitro models with explants of these tissues. Limbal stem cells (LSCs) are already used to treat severe corneal damage as limbal stem cell deficiency. However, this...
|
|
|
Effect of oxidative stress on the differentiation of mesenchymal stem cells.
Bura, Radek ; Tlapáková, Tereza (advisor) ; Pacherník, Jiří (referee)
Mesenchymal stem cells are capable of forming different types of tissue such as muscle, bone, fat or cartilage tissue, thanks to the ability to divide and transform into another cell type. Mesenchymal stem cells obtained from various tissues are used for cell therapy and regenerative medicine. Knowledge of the influence of various factors on the differentiation of these multipotent cells is important. Currently, little is known about the effect of reactive oxygen species (ROS) on the differentiation potential of mesenchymal stem cells. The aim of this bachelor's thesis is to summarize the knowledge of the effect of oxidative stress on the differentiation of mesenchymal stem cells and the affected signaling pathways.
|
guest ::
