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The use of stem cells in the experimental model of stroke
Turnovcová, Karolína ; Jendelová, Pavla (advisor) ; Bojar, Martin (referee) ; Mazurová, Yvona (referee)
Human stem cells from diverse origin are a most promising source for innovative therapy in acute brain lesions. Here, we evaluated the potential of human pluripotent cell-derived neural precursor therapy in stroke, we studied growth properties and surface marker expression of human mesenchymal stromal cells cultivated in different media and introduced superparamagnetic iron oxide nanoparticles for intracellular labeling and noninvasive tracking of transplanted cells. Our results showed that human embryonic cells and human induced pluripotent cells are able to differentiate towards transplantable neural and neuronal precursors. Our cells can follow the neuroectodermal development described in brain and spinal cord tissue during ontogenesis, which is characterized by the expression of different surface and cytoplasmic markers appearing on distinct levels of neurogenesis. Based on this expression, we defined our pluripotent cell-derived neural precursors as neural stem cells and neural progenitors and defined the most suitable developmental level for neural transplantation. We found a double effect of these transplants in restoring neurological functions; firstly, the neural transplants have a paracrine effect on damaged tissue, which is rapid and transient and, secondly, they have an effect on the...
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Neurogeneze a gliogeneze v dospělém mozku po ischemickém poškození
Honsa, Pavel ; Anděrová, Miroslava (advisor) ; Vyklický, Ladislav (referee) ; Mazurová, Yvona (referee)
Ischemic brain injury belongs to the most common cause of death in the developed countries. High complexity of this disorder significantly slows and limits the possible treatment. Currently, there is only one treatment available - the application of the thrombolytic, tissue plasminogen activator, while thousands of other drugs failed during clinical testing. Great expectations were seen in the stroke treatment employing neural stem cells obtained from several external sources; nevertheless, low survival rate, limited favorable outcome and enormous ethical issues complicate the application of such therapy. On the other hand, in the adult mammalian brain exist two endogenous processes - neurogenesis and gliogenesis. These processes need to be fully described and understood, in order to employ them as a source of new cells after injury. Therefore, this thesis focuses on the processes of adult neurogenesis and gliogenesis predominantly after ischemia. Adult neurogenesis and gliogenesis are processes, by which neurons or glial cells are generated from stem/progenitor cells. Both these processes are strongly influenced by brain injury; nevertheless, their contribution to regeneration after ischemia in the human brain is negligible. Here, we aimed to describe the role of polydendrocytes in the...
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Membrane properties of neural stem/progenitor cells during in vitro differentiation and after transplantation into the rat brain
Prajerová, Iva ; Chvátal, Alexandr (advisor) ; Vyklický, Ladislav (referee) ; Mazurová, Yvona (referee)
AOSTRACT The aim oflhis .....orl:. was to elucidate the dilTercntiation mechanisms of neural stcmlpmgenitor cells in vitro and their potentiallO survive and ditfcn:ntiate in \'il'o, al\er trarlSplnntatioo into thc injured mt conex. IImnunohistochcmistI)' "'lIS used for cell idcntiticatioos. and lhe p'0pCl ties ofK' aod Na' voltagc-gated ion ehnnnels were studiC<! using the pat,h-(:Iamp teclmique, We Ilave demonstrnted lhal inunortalised green lluort:SCCnll)r()lein (GFP)INE-4C neural stem cells derivtXI fmm tbc neurocpithelium of p5l'<lctieicm mouse cmbryos iII cmbryonic day (E)9 are ablc to dilferentiate i"to !leUrQns in \1"0, After tmnsplantalioo illto the site of a photlxhcmical lC'iion of aduh rol5, GFP!NE4C cdls slIn'ive and give Tise to neurons. astrocytt5 and oHgodmdrocytcs. Prima!)' embryonic ncura.l Slem cells were isoLaled frum O6IGFP ntice, in which GFP is expressed un(\cr tbc IXlfItrol of06. 11 promoter of thc mDachl gene. which is in\'Olved in tbe development of Ihc concx. At E1 2. D6 is specifically exprcssed in loc neurd.! stem cclLs of the (\(nal tclcnocephalon. from whieh cortical neurons arise, We have shown lila! D61GFP neurJI stem odLs isolated fmm 1312 embfyos are able to give rlse to neuroos and glial fibrillary acidic protcin (GFAP)-posilivc cel1s ill \'i"o nnd that after...
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Morphological changes of cardiopulmonary vascular bed under experimental conditions
Novotný, Tomáš ; Vajner, Luděk (advisor) ; Mazurová, Yvona (referee) ; Tonar, Zbyněk (referee)
In rats, the environment with low content of oxygen induces the development of hypoxic pulmonary hypertension based also on remodeling of pulmonary resistance arteries. This process is particularly triggered by the mast cell degranulation products, especially rodent-like interstitial collagenase (matrix metalloproteinase 13). Sodium cromoglycate administration leads to an effective stabilization of the mast cell granules and thus to the modified remodeling process. During 4-day and 21-day hypoxia, we treated Wistar Han rats with sodium cromoglycate in case control study. We assessed pulmonary vascular remodeling morphology using conventional histological techniques and immunohistochemistry. Then we assessed counts of pulmonary mast cells, both total and MMP-13 positive ones, around pulmonary arteries. Hypoxia induced remodeling of all categories of pulmonary arteries. Sodium cromoglycate treatment in first four days of hypoxia modified and reduced these changes. Treatment in last four days of 21-days hypoxia experiment has only negligible effect. Hypoxia led to changes in collagen VI amount and distribution as well as decrease in number of toluidine blue detectable mast cell as well as MMP13 positive mast cells in the wall of pulmonary arteries. Hypoxia led to significant remodeling of all...
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Morphological changes of cardiopulmonary vascular bed under experimental conditions
Novotný, Tomáš ; Vajner, Luděk (advisor) ; Mazurová, Yvona (referee) ; Tonar, Zbyněk (referee)
In rats, the environment with low content of oxygen induces the development of hypoxic pulmonary hypertension based also on remodeling of pulmonary resistance arteries. This process is particularly triggered by the mast cell degranulation products, especially rodent-like interstitial collagenase (matrix metalloproteinase 13). Sodium cromoglycate administration leads to an effective stabilization of the mast cell granules and thus to the modified remodeling process. During 4-day and 21-day hypoxia, we treated Wistar Han rats with sodium cromoglycate in case control study. We assessed pulmonary vascular remodeling morphology using conventional histological techniques and immunohistochemistry. Then we assessed counts of pulmonary mast cells, both total and MMP-13 positive ones, around pulmonary arteries. Hypoxia induced remodeling of all categories of pulmonary arteries. Sodium cromoglycate treatment in first four days of hypoxia modified and reduced these changes. Treatment in last four days of 21-days hypoxia experiment has only negligible effect. Hypoxia led to changes in collagen VI amount and distribution as well as decrease in number of toluidine blue detectable mast cell as well as MMP13 positive mast cells in the wall of pulmonary arteries. Hypoxia led to significant remodeling of all...
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The use of stem cells in the experimental model of stroke
Turnovcová, Karolína ; Jendelová, Pavla (advisor) ; Bojar, Martin (referee) ; Mazurová, Yvona (referee)
Human stem cells from diverse origin are a most promising source for innovative therapy in acute brain lesions. Here, we evaluated the potential of human pluripotent cell-derived neural precursor therapy in stroke, we studied growth properties and surface marker expression of human mesenchymal stromal cells cultivated in different media and introduced superparamagnetic iron oxide nanoparticles for intracellular labeling and noninvasive tracking of transplanted cells. Our results showed that human embryonic cells and human induced pluripotent cells are able to differentiate towards transplantable neural and neuronal precursors. Our cells can follow the neuroectodermal development described in brain and spinal cord tissue during ontogenesis, which is characterized by the expression of different surface and cytoplasmic markers appearing on distinct levels of neurogenesis. Based on this expression, we defined our pluripotent cell-derived neural precursors as neural stem cells and neural progenitors and defined the most suitable developmental level for neural transplantation. We found a double effect of these transplants in restoring neurological functions; firstly, the neural transplants have a paracrine effect on damaged tissue, which is rapid and transient and, secondly, they have an effect on the...
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Neurogeneze a gliogeneze v dospělém mozku po ischemickém poškození
Honsa, Pavel ; Anděrová, Miroslava (advisor) ; Vyklický, Ladislav (referee) ; Mazurová, Yvona (referee)
Ischemic brain injury belongs to the most common cause of death in the developed countries. High complexity of this disorder significantly slows and limits the possible treatment. Currently, there is only one treatment available - the application of the thrombolytic, tissue plasminogen activator, while thousands of other drugs failed during clinical testing. Great expectations were seen in the stroke treatment employing neural stem cells obtained from several external sources; nevertheless, low survival rate, limited favorable outcome and enormous ethical issues complicate the application of such therapy. On the other hand, in the adult mammalian brain exist two endogenous processes - neurogenesis and gliogenesis. These processes need to be fully described and understood, in order to employ them as a source of new cells after injury. Therefore, this thesis focuses on the processes of adult neurogenesis and gliogenesis predominantly after ischemia. Adult neurogenesis and gliogenesis are processes, by which neurons or glial cells are generated from stem/progenitor cells. Both these processes are strongly influenced by brain injury; nevertheless, their contribution to regeneration after ischemia in the human brain is negligible. Here, we aimed to describe the role of polydendrocytes in the...
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Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain
Němečková, Ivana ; Mazurová, Yvona (advisor) ; Anděrová, Miroslava (referee) ; Martínek, Jindřich (referee)
("Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain") Huntington's disease (HD) is an inherited neurodegenerative disorder. Although the cause of HD, i.e. the production of the mutant form of unstable protein huntingtin (mhtt) which contains 40 and more CAG repeats is known, the effective therapy is not yet available. Therefore, the use of animal models is crucial for the study of the pathogenesis of this fatal disorder. To date, there is no suitable experimental model simulating the neurodegenerative process (NDP) developing in the striatum of the human HD brain. Most of rodent models of HD fall into two broad categories - the neurotoxic lesions and genetically engineered models. The primary aim of our study was a comprehensive morphological description of the development of NDP of HD phenotype in the striatum of the rat brain. We compared the progression of NDP in the lesion induced by intrastriatal injection of quinolinic acid (QA) and in rats transgenic for HD. The groups of male rats surviving for 3, 6-7, 14 days, 1, 3, 6, 9 and 12 months after the QA lesion were compared with 2-, 6-, 12-, 18-, 22-24-month-old tgHD rats and age-matched control (intact) counterparts in both groups. The primary morphological feature of the NDP of HD...
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Membrane properties of neural stem/progenitor cells during in vitro differentiation and after transplantation into the rat brain
Prajerová, Iva ; Chvátal, Alexandr (advisor) ; Vyklický, Ladislav (referee) ; Mazurová, Yvona (referee)
AOSTRACT The aim oflhis .....orl:. was to elucidate the dilTercntiation mechanisms of neural stcmlpmgenitor cells in vitro and their potentiallO survive and ditfcn:ntiate in \'il'o, al\er trarlSplnntatioo into thc injured mt conex. IImnunohistochcmistI)' "'lIS used for cell idcntiticatioos. and lhe p'0pCl ties ofK' aod Na' voltagc-gated ion ehnnnels were studiC<! using the pat,h-(:Iamp teclmique, We Ilave demonstrnted lhal inunortalised green lluort:SCCnll)r()lein (GFP)INE-4C neural stem cells derivtXI fmm tbc neurocpithelium of p5l'<lctieicm mouse cmbryos iII cmbryonic day (E)9 are ablc to dilferentiate i"to !leUrQns in \1"0, After tmnsplantalioo illto the site of a photlxhcmical lC'iion of aduh rol5, GFP!NE4C cdls slIn'ive and give Tise to neurons. astrocytt5 and oHgodmdrocytcs. Prima!)' embryonic ncura.l Slem cells were isoLaled frum O6IGFP ntice, in which GFP is expressed un(\cr tbc IXlfItrol of06. 11 promoter of thc mDachl gene. which is in\'Olved in tbe development of Ihc concx. At E1 2. D6 is specifically exprcssed in loc neurd.! stem cclLs of the (\(nal tclcnocephalon. from whieh cortical neurons arise, We have shown lila! D61GFP neurJI stem odLs isolated fmm 1312 embfyos are able to give rlse to neuroos and glial fibrillary acidic protcin (GFAP)-posilivc cel1s ill \'i"o nnd that after...
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