DISP3 (PTCHD2), a sterol-sensing domain-containing protein, is highly expressed in neural tissue but its role in neural differentiation is unknown. In the present study we used a multipotent cerebellar progenitor cell line, C17.2, to investigate the impact of DISP3 on the proliferation and differentiation of neural precursors. We found that ectopically expressed DISP3 promotes cell proliferation and alters expression of genes that are involved in tumorigenesis. Finally, the differentiation profile of DISP3-expressing cells was altered, as evidenced by delayed expression of neural specific markers and a reduced capacity to undergo neural differentiation.

• MeSH
• buněčná diferenciace * MeSH
• buněčné linie MeSH
• lidé MeSH
• membránové proteiny genetika   metabolismus   MeSH
• metabolismus lipidů MeSH
• mozek cytologie   MeSH
• nervové kmenové buňky cytologie   metabolismus   MeSH
• proliferace buněk MeSH
• regulace genové exprese MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Kmenové buňky pro své jedinečné vlastnosti – plasticitu, slibují všestranné využití, zejména možnostireparace většiny orgánů a tkání lidského těla – buněčná terapie. Autoři ve svémsdělení prezentují svojeexperimentální výsledky na modelu neurálních kmenových buněk (NSCs). Hlavním cílem je průkazzachování hemopoetické identity NSCs. Subletálně celotělově ozářené myši dávkou LD 8,5 Gy bylyzachráněné transplantací značených (X-Gal+) NSCs. Průkaz zvýšeného záchytu NSCs ve slezině násvedlo k myšlence hledání vztahu hemopoetického mikroprostředí a hemopoetické identity. Průkazzachování hemopoetické identity byl proveden pomocí kultivace CFU-GM z kostní dřeně – kde bylyidentifikovány buňky vykazující X-Gal pozitivitu (stanovení těchto buněk bylo provedeno pomocíhistochemického testu). Naše experimenty poukazují na různé možnosti, jak transplantace NSCs můžeovlivnit poškozenou hematopoézu.

Stem cells for their unique property - plasticity promise a universal utilization,mainly, in the reparationof most organs and tissues in the human body. In this communication, the authors presents theirexperimental results in a model of neural stem cells (NCSc) where the main goal was to preserve theNSCs hemopoietic identity. Mice given whole body sublethal irradiation with a dose of LD 8.5 Gy weresaved by transplantation of labelled (X-Gal+) NSCs. The demonstration of increased uptake of NSCs inthe spleen led us to study the relationship of hemopoietic microenvironment and hemopoietic identity.The proof of hemopoeitic identity was performed by cultivation of CFU-GM from the bone marrow -where cells exhibiting X-Gal positivity were identified by histochemistry. Our experiments showdifferent ways how NSCs transplantation may influence the damaged hematopoiesis.

• MeSH
• dávka záření MeSH
• finanční podpora výzkumu jako téma MeSH
• hematopoéza MeSH
• kmenové buňky fyziologie   MeSH
• kostní dřeň MeSH
• kultivované buňky MeSH
• modely u zvířat MeSH
• myši MeSH
• transplantace kostní dřeně MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• přehledy MeSH
• srovnávací studie MeSH

Notch and gp130 signaling are involved in the regulation of multiple cellular processes across various tissues during animal ontogenesis. In the developing nervous system, both signaling pathways intervene at many stages to determine cell fate-from the first neural lineage commitment and generation of neuronal precursors, to the terminal specification of cells as neurons and glia. In most cases, the effects of Notch and gp130 signaling in these processes are similar. The aim of the current review was to summarize the knowledge regarding the roles of Notch and gp130 signaling in the maintenance of neural stem and progenitor cells during animal ontogenesis, from early embryo to adult. Recent data show a direct crosstalk between these signaling pathways that seems to be specific for a particular type of neural progenitors.

• MeSH
• cytokinový receptor gp130 metabolismus   MeSH
• interakce mezi receptory a ligandy MeSH
• lidé MeSH
• nervové kmenové buňky metabolismus   MeSH
• neurogeneze MeSH
• receptory Notch metabolismus   MeSH
• signální transdukce * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Both gain- and loss-of-function mutations have recently implicated HCFC1 in neurodevelopmental disorders. Here, we extend our previous HCFC1 over-expression studies by employing short hairpin RNA to reduce the expression of Hcfc1 in embryonic neural cells. We show that in contrast to over-expression, loss of Hcfc1 favoured proliferation of neural progenitor cells at the expense of differentiation and promoted axonal growth of post-mitotic neurons. To further support the involvement of HCFC1 in neurological disorders, we report two novel HCFC1 missense variants found in individuals with intellectual disability (ID). One of these variants, together with three previously reported HCFC1 missense variants of unknown pathogenicity, were functionally assessed using multiple cell-based assays. We show that three out of the four variants tested result in a partial loss of HCFC1 function. While over-expression of the wild-type HCFC1 caused reduction in HEK293T cell proliferation and axonal growth of neurons, these effects were alleviated upon over-expression of three of the four HCFC1 variants tested. One of these partial loss-of-function variants disrupted a nuclear localization sequence and the resulting protein displayed reduced ability to localize to the cell nucleus. The other two variants displayed negative effects on the expression of the HCFC1 target gene MMACHC, which is responsible for the metabolism of cobalamin, suggesting that these individuals may also be susceptible to cobalamin deficiency. Together, our work identifies plausible cellular consequences of missense HCFC1 variants and identifies likely and relevant disease mechanisms that converge on embryonic stages of brain development.

• MeSH
• aktivní transport - buněčné jádro MeSH
• buněčná diferenciace genetika   MeSH
• exprese genu MeSH
• faktor C1 hostitelské buňky chemie   genetika   metabolismus   MeSH
• HEK293 buňky MeSH
• kultivované buňky MeSH
• lidé MeSH
• malá interferující RNA genetika   MeSH
• mentální retardace genetika   MeSH
• mozek cytologie   embryologie   MeSH
• mutace * MeSH
• myši MeSH
• nervové kmenové buňky cytologie   metabolismus   MeSH
• proliferace buněk MeSH
• RNA interference MeSH
• rodokmen MeSH
• sekvence aminokyselin MeSH
• substituce aminokyselin MeSH
• transdukce genetická MeSH
• transportní proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Neural crest (NC) is a transient embryonic tissue, whose cells are motile and multipotent until they reach their destination and differentiate according to microenvironmental cues into a variety of cell types. However, a subpopulation of these cells remains multipotent. They were found, among other locations, in a bulge of adult murine whisker follicle and were designated epidermal neural crest stem cells (EPI-NCSCs). The aim of this work is to ascertain whether the EPI-NCSCs could be isolated from human hair follicles as well. Due to their exceptional properties, they could represent potential candidates for stem cell therapy. The presented work focuses on the isolation and characterization of EPI-NCSCs from human skin. We obtained a population of cells that expressed markers of NC, NC progeny and general stem cell markers. After prolonged cultivation, the subpopulation of cells spontaneously differentiated into some of NC derivatives, i.e. neurons, smooth muscle cells and Schwann cell progenitors. Targeted differentiation with neuregulin 1 highly increased the number of Schwann cells in the culture. Human EPI-NCSCs could also grow under non-adherent conditions and form 3-dimensional spheres. Microarray analysis was performed and gene profile of human EPI-NCSCs was compared with the list of key genes of murine EPI-NCSCs and the list of genes up-regulated in newly induced NC cells. This revealed 94% and 88% similarity, respectively. All presented results strongly support the NCSC identity and multipotency of isolated human cells. These cells could thus be used in regenerative medicine, especially because of the easy accessibility of donor tissue.

• MeSH
• buněčná diferenciace MeSH
• crista neuralis cytologie   metabolismus   MeSH
• financování organizované MeSH
• imunohistochemie MeSH
• kmenové buňky cytologie   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• multipotentní kmenové buňky cytologie   metabolismus   MeSH
• myši MeSH
• neurony MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• Schwannovy buňky MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• stanovení celkové genové exprese MeSH
• transplantace kmenových buněk MeSH
• vibrissae MeSH
• vlasový folikul cytologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH

The isolation of neural stem cells from fetal and adult mammalian CNS and the demonstration of functional neurogenesis in adult CNS have offered perspectives for treatment of many devastating hereditary and acquired neurological diseases. Due to this enormous potential, neural stem cells are a subject of extensive molecular profiling studies with a search for new markers and regulatory pathways governing their self-renewal as opposed to differentiation. Several in-depth proteomic studies have been conducted on primary or immortalized cultures of neural stem cells and neural progenitor cells, and yet more remains to be done. Additionally, neurons and glial cells have been obtained from embryonic stem cells and mesenchymal stem cells, and proteins associated with the differentiation process have been characterized to a certain degree with a view to further investigations. This review summarizes recent findings relevant to the proteomics of neural stem cells and discusses major proteins significantly regulated during neural stem cell differentiation with a view to their future use in cell-based regenerative and reparative therapy.

• MeSH
• buněčná diferenciace MeSH
• financování organizované MeSH
• kmenové buňky cytologie   chemie   MeSH
• lidé MeSH
• neurony cytologie   chemie   MeSH
• proteiny fyziologie   genetika   MeSH
• proteomika MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Despite therapeutic advances, neurodegenerative diseases and disorders remain some of the leading causes of mortality and morbidity in the United States. Therefore, cell-based therapies to replace lost or damaged neurons and supporting cells of the central nervous system (CNS) are of great therapeutic interest. To that end, human pluripotent stem cell (hPSC) derived neural progenitor cells (hNPCs) and their neuronal derivatives could provide the cellular 'raw material' needed for regenerative medicine therapies for a variety of CNS disorders. In addition, hNPCs derived from patient-specific hPSCs could be used to elucidate the underlying mechanisms of neurodegenerative diseases and identify potential drug candidates. However, the scientific and clinical application of hNPCs requires the development of robust, defined, and scalable substrates for their long-term expansion and neuronal differentiation. In this study, we rationally designed a vitronectin-derived peptide (VDP) that served as an adhesive growth substrate for the long-term expansion of several hNPC lines. Moreover, VDP-coated surfaces allowed for the directed neuronal differentiation of hNPC at levels similar to cells differentiated on traditional extracellular matrix protein-based substrates. Overall, the ability of VDP to support the long-term expansion and directed neuronal differentiation of hNPCs will significantly advance the future translational application of these cells in treating injuries, disorders, and diseases of the CNS.

• MeSH
• biokompatibilní potahované materiály farmakologie   MeSH
• buněčná adheze účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• extracelulární matrix - proteiny metabolismus   MeSH
• lidé MeSH
• molekuly buněčné adheze metabolismus   MeSH
• myši MeSH
• nervové kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• neurony cytologie   účinky léků   metabolismus   MeSH
• peptidy farmakologie   MeSH
• pluripotentní kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• vitronektin farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Among various strategies employed for spinal cord injury, stem cell therapy is a potential treatment. So far, a variety of stem cells have been evaluated in animal models and humans with spinal cord injury, and epidermal neural crest stem cells represent one of the attractive types in this area. Although these multipotent stem cells have been assessed in several spinal cord injury models by independent laboratories, extensive work remains to be done to ascertain whether these cells can safely improve the outcome following human spinal cord injury. Among the models that closely mimic human spinal cord injury, the in vitro model of injury in organotypic spinal cord slice culture has been identified as one of the faithful platforms for injury-related investigations. In this study, green fluorescent protein-expressing stem cells were grafted into injured organotypic spinal cord slice culture and their survival was examined by confocal microscope seven days after transplantation. Data obtained from this preliminary study showed that these stem cells can survive on top of the surface of injured slices, as observed on day seven following their transplantation. This result revealed that this in vitro model of injury can be considered as a suitable context for further evaluation of epidermal neural crest stem cells before their application in large animals.

• MeSH
• biologické modely MeSH
• buněčná smrt MeSH
• crista neuralis cytologie   MeSH
• epidermální buňky MeSH
• epidermis MeSH
• kmenové buňky cytologie   MeSH
• kultivované buňky MeSH
• mícha cytologie   MeSH
• potkani Wistar MeSH
• transplantace kmenových buněk MeSH
• tvar buňky MeSH
• viabilita buněk MeSH
• zelené fluorescenční proteiny metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Traumatic spinal cord injury (SCI) triggers a chain of events that is accompanied by an inflammatory reaction leading to necrotic cell death at the core of the injury site, which is restricted by astrogliosis and apoptotic cell death in the surrounding areas. Activation of nuclear factor-κB (NF-κB) signaling pathway has been shown to be associated with inflammatory response induced by SCI. Here, we elucidate the pattern of activation of NF-κB in the pathology of SCI in rats and investigate the effect of transplantation of spinal neural precursors (SPC-01) on its activity and related astrogliosis. METHODS: Using a rat compression model of SCI, we transplanted SPC-01 cells or injected saline into the lesion 7 days after SCI induction. Paraffin-embedded sections were used to assess p65 NF-κB nuclear translocation at days 1, 3, 7, 10, 14, and 28 and to determine levels of glial scaring, white and gray matter preservation, and cavity size at day 28 after SCI. Additionally, levels of p65 phosphorylated at Serine536 were determined 10, 14, and 28 days after SCI as well as levels of locally secreted TNF-α. RESULTS: We determined a bimodal activation pattern of canonical p65 NF-κB signaling pathway in the pathology of SCI with peaks at 3 and 28 days after injury induction. Transplantation of SCI-01 cells resulted in significant downregulation of TNF-α production at 10 and 14 days after SCI and in strong inhibition of p65 NF-κB activity at 28 days after SCI, mainly in the gray matter. Moreover, reduced formation of glial scar was found in SPC-01-transplanted rats along with enhanced gray matter preservation and reduced cavity size. CONCLUSIONS: The results of this study demonstrate strong immunomodulatory properties of SPC-01 cells based on inhibition of a major signaling pathway. Canonical NF-κB pathway activation underlines much of the immune response after SCI including cytokine, chemokine, and apoptosis-related factor production as well as immune cell activation and infiltration. Reduced inflammation may have led to observed tissue sparing. Additionally, such immune response modulation could have impacted astrocyte activation resulting in a reduced glial scar.

• MeSH
• časové faktory MeSH
• cytokiny metabolismus   MeSH
• gliový fibrilární kyselý protein metabolismus   MeSH
• glióza chirurgie   MeSH
• kmenové buňky fyziologie   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• poranění míchy komplikace   MeSH
• potkani Wistar MeSH
• signální transdukce fyziologie   MeSH
• transformované buněčné linie MeSH
• transkripční faktor RelA metabolismus   MeSH
• transplantace kmenových buněk metody   MeSH
• zánět etiologie   chirurgie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• kmenové buňky MeSH
• tkáňové inženýrství MeSH
• výzkum embrya etika   MeSH
• výzkum plodu etika   MeSH
• Publikační typ
• příručky MeSH

• Konspekt
• Biotechnologie. Genetické inženýrství
• NLK Obory
• embryologie a teratologie
• biomedicínské inženýrství