|
|
Local production of cytokines after treatment with stem cells of damaged ocular surface
Kössl, Jan ; Holáň, Vladimír (advisor) ; Kovář, Marek (referee)
The damage of ocular surface represents one of the most common causes of decreased quality of vision or even blindness. If the injury is extensive and includes the region of limbus, niche of limbal stem cells (LSC), LSC deficiency occurs and the natural corneal regeneration is stopped. Conjunctival epithelium migrates into the injured area. Neovascularization, local inflammation and corneal opacity occur. Corneal transplantation is an insufficient treatment in such case. If the injury is bilateral, the allogenic limbal graft or LSC transplantation is required. In such cases systemic immunosuppressive drugs with many negative side-effects must be administered. The search for an adequate autologous substitution is important for avoid immunosuppressive medication. Mesenchymal stem cells (MSC) represent a perspective substitution for the reason of their immunomodulatory properties and the capability to differentiate in many cell types. There is possibility to isolate sufficient number of these cells from adipose tissue or bone marrow which are relatively easily accessible. Our goal was to observe local production of cytokines and other molecules which are present in inflammatory reaction after the chemical burn of the murine cornea and after the treatment with stem cells growing on nanofiber scaffold....
|
|
|
Immunomodulatory properties of mesenchymal stem cells from patients with amyotrophic lateral sclerosis and healthy donors
Matějčková, Nicole ; Javorková, Eliška (advisor) ; Kanderová, Veronika (referee)
Mesenchymal stem cells (MSC) possess a multilineage differentiation potential and have the ability to regulate reactivity of the immune system. They are usually isolated and expanded from the bone marrow, adipose tissue or umbilical cord. MSC represent promising cell population for the treatment of some severe diseases, such as amyotrofic lateral sclerosis (ALS), due to the combination of regenerative and immunomodulatory properties. The aim of this study is to compare MSC from ALS patients and healthy donors in their phenotype, proliferative activity and mainly their immunomodulatory properties. The assessment of impact of the disease on the properties of MSC is important for their autologous use in clinical trials. In this study we used MSC isolated from bone marrow of 14 ALS patients and 15 patients undergoing mostly orthopedic surgery as control group. We also used MSC stimulated for 24 hours by poinflammatory cytokines. Cells were compared in terms of immunophenotype, differentiation in adipocytes and osteoblasts, metabolic activity, expression of selected genes for immunomodulatory molecules and for inhibition of lymphocyte proliferation. Further experiments were focused on evaluation of immunomodulatory properties of MSC. The effect of MSC on peripheral blood mononuclear cells stimulated...
|
|
|
Effect of growth factors and bioactive substances on mesenchymal stem cell differentiation.
Blahnová, Veronika ; Daňková, Jana (advisor) ; Zajícová, Alena (referee)
Mesenchymal stem cells are a population of multipotent cells, which have the ability to restore damaged tissues derived from mesoderm. Under the influence of wide range of growth factors, hormones and other bioactive molecules they can differentiate for example into chondrocytes, osteocytes, adipocytes, myocytes or insulin producing cells. The differentiation is induced even by contacts with neighboring cells or with extracellular matrix. Tissue engineering often uses especially growth factors. Growth factors act through specific surface receptors, which mediate cell-to-cell and cell-to-extracelular matrix communication and influence many cellular processes such as growth, proliferation, differentiation and others. Due to their specific impact growth factors are used individually or in combinations in tissue engineering applications. Substances stimulating cells to proliferate and differentiate can be added right to the culture medium, when cultivated in vitro, or can be loaded into a scaffold onto which cells are later seeded. The determination of exact growth factors combinations and concentrations influence on MSCs differentiation would enable more efficient use of MSCs in regenerative medicine. The aim of this thesis is to summarise present knowledge about the effect of growth factors and other...
|
|
|
Study of effect of immunosuppressive drugs on interaction of mesenchymal stem cells with immune cells
Heřmánková, Barbora ; Krulová, Magdaléna (advisor) ; Indrová, Marie (referee)
Mesenchymal stem cells (MSC) represent a heterogenous population of nonhematopoietic stem cells with multipotent differential potential. MSC can be isolated from various tissues of organism, the most common tissue are bone marrow or adipose tissue. MSC are good candidates for treatment of autoimmune diseases and possess the ability to prevent graft rejection or graft versus host disease. The immunosuppressive drugs are currently used for inhibition of unwanted immune reaction but they exhibit serious side effects. The use of MSC in therapy can reduce doses of immunosuppressive drugs and eliminate side effects. The study of MSC and immunosuppressant interactions should be detected before MSC can be used for clinical application. We aimed to analyze the interaction between MSC and immunosuppressive drugs and their effect on immune cells. Cyclosporine A and mycophenolate mofetil were used in our research. We demonstrated changes in the expression of surface molecules, production of interleukin 6 and in metabolic activity of MSC after treatment with immunosuppressive drugs. MSC are in organism, in cooperation with the number of other cells. Therefore we studied MSC cocultured with splenocytes in the presence of immunosuppressive drugs. Our results show the effect of MSC and immunosuppressive drugs on different...
|
|
|
The use of immunoregulatory properties of mesenchymal stem cells/ and their therapeutic potential
Javorková, Eliška ; Krulová, Magdaléna (advisor) ; Stříž, Ilja (referee) ; Tučková, Ludmila (referee)
Mesenchymal stem cells (MSCs) have the potential to differentiate into various cell types, possess potent immunomodulatory properties and can influence various functions of immune cells. Since the immunomodulatory properties of MSCs can be modified by cytokines, we compered the effect of unstimulated MSCs and MSCs pretreated with interleukin (IL)-1, interferon (IFN)- , transforming growth factor (TGF)- and IL-10 on the development of regulatory T cells (Treg) and T helper 17 (Th17) cells in vitro and on the inflammatory environment in the eye. MSCs can produce significant levels of TGF- and IL-6. These cytokines represent the key factors that reciprocally regulate the development of naive T cells into Treg and Th17 cells. Unstimulated MSCs produce TGF- , but not IL-6, and the production of TGF- can be further enhanced by IL-10 or TGF- . In the presence of IL-1, MSCs secrete significant levels of IL-6, in addition to spontaneous production of TGF- . MSC producing TGF- induced preferentially expression of Foxp3 and activation of Treg lymphocytes, whereas MSCs supernatants containing TGF- together with IL-6 supported ROR t expression and development of Th17 cells. We demonstrated that MSCs and their products effectively control the development of Tregs and Th17 cells in a population of...
|
|
|
Effect of the peptides on osteogenic differentiation of mesenchymal steam cells
Lukášová, Věra ; Amler, Evžen (advisor) ; Filová, Elena (referee)
Osteogenic differentiation of mesenchymal stem cells (MSCs) would be possible to induce by creating of a cell bioactive scaffold that mimic the properties of bone extracellular matrix (ECM). This induction will be not only due to the addition of osteogenic supplements, but also due to the addition of differentiation peptides. These peptides activate signaling pathways leading to cell differentiation. The aim of this study was to evaluate the effect of selected peptides on adhesion, metabolic activity, proliferation and osteogenic differentiation of porcine MSCs. Four peptides with amino acid sequences of DGEA, IAGVGGEKSGGF, GQGFSYPYKAVFSTQ and KIPKASSVPTELSAISTLYL were selected. These peptides were derived from receptor binding sequences of collagen I, collagen III, BMP-7 and BMP-2 respectively. Scaffolds were prepared from a biocompatible and biodegradable poly-ε-caprolactone (PCL) polymer, suitable for cell cultivation. Cells were cultured on scaffolds for three weeks. Various concentration of differentiation peptides were added to the culture medium. As observed in the experiment of cells cultured in basal medium supplemented with differentiation peptides no effect on adhesion, proliferation or metabolic activity of porcine MSCs was observed. In groups treated with peptides derived from BMP-2...
|
|
|
Treatment of spinal cord injury by transplantation different types of stem cells
Dubišová, Jana ; Kubinová, Šárka (advisor) ; Cizkova, Dasa (referee)
Spinal cord injury (SCI) is complicated injury with serious socioeconomic consequences for the patient and his whole family. Big difficulty cause also extremely high living expenses for the patient with this type of injury. That's why there is a need for therapeutic methods which would help patients after SCI to recover the lost functions and be able at least partially to return to their normal life. Different therapeutic methods are being used for SCI treatment. In this study we used four various types of stem cells: human bone marrow stem cells (hBM-MSCs), human umbilical cord mesenchymal stem cells (hUC-MSCs), neural precursors derived from induced pluripotent stem cells (iPS-NPs) and neural stem cell line derived from human fetal spinal cord tissue (SPC-01). These cells have been transplanted intrathecally or intraspinally 7 days after induction of the experimental model of SCI in the rat. We studied expressions of genes related to neurogenesis, growth factors and inflammation 10 and 28 days after SCI. Our analysis showed significant changes in gene expression 10 days after SCI. Significant up-regulation in expression of vascular endothelial growth factor (Vegf), ciliary neurotrophic factor (Cntf) and interferon regulatory factor 5 (Irf5) were found after transplantation of hBM-MSCs and hUC-...
|
|
|
Mesenchymal stem cells for treatment of spinal cord injury
Dubišová, Jana ; Kubinová, Šárka (advisor) ; Hock, Miroslav (referee)
Poranenie miechy a jeho terapia pomocou mezenchymálnych kmeňových buniek Abstract Patients with serious spinal cord injury (SCI) have a little chance of a full return to their original life. This kind of injuries leaves lifelong consequences as on the patients themselves, as well as their surroundings and family. Considerable financial resources are, worldwide, granted for reintegratation of these patients into society. This work focuses on the description of spinal cord injury, its complexity and the possibility of treatment by new therapeutic methods of mesenchymal cells. The first section discusses the severity of spinal cord injury and its pathophysiology with the performance of subsequent clinical phases. The second part is dedicated to new treatment methods and approaches, with an emphasis on cell therapy. The third part presents treatment method using mesenchymal stem cells (MSCs), which immunomodulatory and anti-inflammatory properties, such as their ability to migration to the injured area and the possibility of genetic modification, made them a great candidates for the treatment of serious injuries. Keywords Spinal cord injury, stem cells, mesenchymal stem cells, cell therapy, regenerative medicine
|
|
|
Use of biological materials for tissue substitution in plastic surgery
Měšťák, Ondřej ; Sukop, Andrej (advisor) ; Brychta, Pavel (referee) ; Dražan, Luboš (referee)
Užití biologických materiálů k náhradě tkání v plastické chirurgii ! Abstrakt v angličtině Background: Biological meshes are biomaterials consisted of extracellular matrix and used in surgery particularly for hernia treatment or thoracic wall reconstruction. They are capable of vascularization, that decreases risk of infection, expecially when used in contaminated fields. This study compared the strength of incorporation and biocompatibility of two porcine-derived grafts (cross-linked and non-cross-linked) in a rat hernia model. In addition, we hypothesized that combination of extracellular matrices with autologous mesenchymal stem cells used for hernia repair would result in increased vascularization and increased strength of incorporation. Methods: Standardized 2 x 4 cm fascial defect was created in 42 Wistar rats and repaired with a cross-linked or a non-cross-linked graft either enriched or non-enriched with stem cells. The rats were sacrificed 3, 6 and 12 months later. The strength of incorporation, vascularization, cellular invasion, foreign body reaction and capsule formation were evaluated. Results: Comparison of stem cell enriched and non-enriched groups showed no significant differences in the capsule thickness, foreign body reaction, cellularization or vascularization. In the non-cross-linked...
|
|
|
Possibilities of using stem cells for treatment of ocular surface injuries
Kössl, Jan ; Holáň, Vladimír (advisor) ; Drbal, Karel (referee)
Ocular surface defects are one of the most common causes of impaired vision or even blindness. Corneal transplantation is the first choice of the treatment of these defects. If the damage is extensive and includes the limbus, niche of limbal stem cells (LSCs), LSC deficiency (LSCD) occurs and reparation with regeneration of cornea is impaired. The only way to treat LSCD is limbal transplantation or transplantation of autologous LSCs from the healthy eye. In cases of bilateral LSCD there are no autologous LSCs available. The use of allogeneic LSCs is associated with the requirement of systemic or local administration of immunosuppressive drugs which have often negative side-effects and outcomes remain uncertain. The alternative for treatment of ocular surface defects and LSCD is finding a new source of an appropriate autologous stem cell substitute which are e.g. mesenchymal stem cells (MSCs). These cells can be obtained from bone marrow or adipose tissue of the particular patient. MSCs can be easily cultivated ex vivo and can be transferred onto the damaged ocular surface using appropriate scaffold. Here they can differentiate to corneal epithelial cells, have immunomodulatory properties and produce numerous trophic and growth factors. The experiments with MSCs on animal models of mechanically or...
|
guest ::
