Neural progenitors
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The cytocompatibility of cardiomyocytes derived from embryonic stem cells and neural progenitors, which were seeded on the surface of composite films made of graphene oxide (GO) and polypyrrole (PPy-GO) or poly(3,4-ethylenedioxythiophene) (PEDOT-GO) are reported. The GO incorporated in the composite matrix contributes to the patterning of the composite surface, while the electrically conducting PPy and PEDOT serve as ion-to-electron transducers facilitating electrical stimulation/sensing. The films were fabricated by a simple one-step electropolymerization procedure on electrically conducting indium tin oxide (ITO) and graphene paper (GP) substrates. Factors affecting the cell behaviour, i.e. the surface topography, wettability, and electrical surface conductivity, were studied. The PPy-GO and PEDOT-GO prepared on ITO exhibited high surface conductivity, especially in the case of the ITO/PPy-GO composite. We found that for cardiomyocytes, the PPy-GO and PEDOT-GO composites counteracted the negative effect of the GP substrate that inhibited their growth. Both the PPy-GO and PEDOT-GO composites prepared on ITO and GP significantly decreased the cytocompatibility of neural progenitors. The presented results enhance the knowledge about the biological properties of electroactive materials, which are critical for tissue engineering, especially in context stimuli-responsive scaffolds.
- MeSH
- bicyklické sloučeniny heterocyklické chemie MeSH
- elektrická vodivost * MeSH
- elektrochemie * MeSH
- grafit farmakologie MeSH
- kardiomyocyty cytologie účinky léků MeSH
- myši MeSH
- nervové kmenové buňky cytologie účinky léků MeSH
- neurogeneze účinky léků MeSH
- polymery chemie farmakologie MeSH
- pyrroly chemie MeSH
- voda chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Práca opisuje možnosť izolácie a separácie neurálnych proge- nitorov získaných z postnatálnej miechy potkanov. Neurálne progenitory sú multipotentné bunky, ktoré sa môžu izolovať z tkanív CNS embryí ako aj dospelých jedincov a v súèasnej dobe predstavujú jeden z možných spôsobov liečby poškodenia centrálneho nervového systému. Výsledky, ktoré sme získali pri využití magnetickej separácie miechových progenitorov ako jednej z izolačných metód pri získavaní populácie buniek obohatených o líniu oligoden- drálnych buniek (80 % pri populácii buniek s magnetickou separáciou vs. 30 % bez magneticke separácie) svedčí o vhodnosti využitia danej separaènej metódy pri izolácii a kultivácii oligodendrocytovej línie neurálnych progenitorov.
This study was designed to describe the isolation and separation possibilty of neural progenitors isolated from postnatal rat spinal cord. Neural progenitors are multipotent cells, which can be iso- lated from CNS embryonic tissue, as well as from adults tissue and at the present time represent one of the potential way for therapeutic approaches in central nervous system injury. Results obtained in this study using magnetic separation for spinal cord progenitors purification confirmed that the magnetic separation is suitable for obtaining and cultivation of oligodendrocytes lineaged neural progenitors (80% in cell population with magnetic separation vs. 30% without magnetic separation).
- Klíčová slova
- neurálne progenitory, oligodendrocyty, magnetická separácia,
- Publikační typ
- abstrakty MeSH
A promising therapeutic strategy for amyotrophic lateral sclerosis (ALS) treatment is stem cell therapy. Neural progenitors derived from induced pluripotent cells (NP-iPS) might rescue or replace dying motoneurons (MNs). However, the mechanisms responsible for the beneficial effect are not fully understood. The aim here was to investigate the mechanism by studying the effect of intraspinally injected NP-iPS into asymptomatic and early symptomatic superoxide dismutase (SOD)1G93A transgenic rats. Prior to transplantation, NP-iPS were characterized in vitro for their ability to differentiate into a neuronal phenotype. Motor functions were tested in all animals, and the tissue was analyzed by immunohistochemistry, qPCR, and Western blot. NP-iPS transplantation significantly preserved MNs, slowed disease progression, and extended the survival of all treated animals. The dysregulation of spinal chondroitin sulfate proteoglycans was observed in SOD1G93A rats at the terminal stage. NP-iPS application led to normalized host genes expression (versican, has-1, tenascin-R, ngf, igf-1, bdnf, bax, bcl-2, and casp-3) and the protection of perineuronal nets around the preserved MNs. In the host spinal cord, transplanted cells remained as progenitors, many in contact with MNs, but they did not differentiate. The findings suggest that NP-iPS demonstrate neuroprotective properties by regulating local gene expression and regulate plasticity by modulating the central nervous system (CNS) extracellular matrix such as perineuronal nets (PNNs).
- MeSH
- amyotrofická laterální skleróza terapie MeSH
- indukované pluripotentní kmenové buňky cytologie MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- nervové kmenové buňky cytologie metabolismus transplantace MeSH
- neuroplasticita * MeSH
- neurotrofní faktory genetika metabolismus MeSH
- periferní nervy fyziologie MeSH
- potkani Sprague-Dawley MeSH
- proteiny regulující apoptózu genetika metabolismus MeSH
- regenerace nervu MeSH
- tenascin genetika metabolismus MeSH
- transplantace kmenových buněk metody MeSH
- versikany genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Neural progenitors of the mouse forebrain can be propagated in vitro as neurospheres in the presence of bFGF and EGF. However, less is understood whether regional characteristics or developmental stage properties of these cells are maintained in neurosphere cultures. Here we show that the original cell fate is lost in neurosphere cultures. We isolated neural progenitors from the dorsal telencephalon of D6-GFP mice and cultured them in vitro. The expression profile was specifically changed in cultured cells in just three passages. Markers of the dorsal forebrain were downregulated and several ventrally-expressed genes were induced. The altered gene expression led to a profound phenotypic change of cultured cells. D6-GFP positive cortical progenitors produce excitatory neurons in the cortex and few astrocytes in vivo but after culture in vitro, these cells differentiate into many astrocytes and also oligodendrocytes and inhibitory neurons. Wnt signaling in cultured neurospheres was downregulated in the same manner as other dorsal markers but dominant active Wnt signaling slowed down the loss of the dorsal identity in neurospheres.
- MeSH
- astrocyty cytologie metabolismus MeSH
- buněčná diferenciace fyziologie MeSH
- buněčné sféroidy cytologie MeSH
- buněčný rodokmen fyziologie MeSH
- financování organizované MeSH
- genetické markery MeSH
- interneurony cytologie metabolismus MeSH
- kmenové buňky cytologie metabolismus MeSH
- mozková kůra cytologie embryologie metabolismus MeSH
- myši transgenní MeSH
- myši MeSH
- neuroglie cytologie metabolismus MeSH
- neurony cytologie metabolismus MeSH
- oligodendroglie cytologie metabolismus MeSH
- promotorové oblasti (genetika) imunologie MeSH
- rekombinantní fúzní proteiny genetika MeSH
- telencefalon cytologie embryologie metabolismus MeSH
- vývojová regulace genové exprese genetika MeSH
- zelené fluorescenční proteiny genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
Spinal cord injury (SCI), is a devastating condition leading to the loss of locomotor and sensory function below the injured segment. Despite some progress in acute SCI treatment using stem cells and biomaterials, chronic SCI remains to be addressed. We have assessed the use of laminin-coated hydrogel with dual porosity, seeded with induced pluripotent stem cell-derived neural progenitors (iPSC-NPs), in a rat model of chronic SCI. iPSC-NPs cultured for 3 weeks in hydrogel in vitro were positive for nestin, glial fibrillary acidic protein (GFAP) and microtubule-associated protein 2 (MAP2). These cell-polymer constructs were implanted into a balloon compression lesion, 5 weeks after lesion induction. Animals were behaviorally tested, and spinal cord tissue was immunohistochemically analyzed 28 weeks after SCI. The implanted iPSC-NPs survived in the scaffold for the entire experimental period. Host axons, astrocytes and blood vessels grew into the implant and an increased sprouting of host TH+ fibers was observed in the lesion vicinity. The implantation of iPSC-NP-LHM cell-polymer construct into the chronic SCI led to the integration of material into the injured spinal cord, reduced cavitation and supported the iPSC-NPs survival, but did not result in a statistically significant improvement of locomotor recovery.
- MeSH
- buněčná diferenciace MeSH
- chronická nemoc MeSH
- hydrogely MeSH
- indukované pluripotentní kmenové buňky metabolismus MeSH
- krysa rodu rattus MeSH
- nervové kmenové buňky transplantace MeSH
- poranění míchy terapie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Schwann cell precursors (SCPs) are nerve-associated progenitors that can generate myelinating and non-myelinating Schwann cells but also are multipotent like the neural crest cells from which they originate. SCPs are omnipresent along outgrowing peripheral nerves throughout the body of vertebrate embryos. By using single-cell transcriptomics to generate a gene expression atlas of the entire neural crest lineage, we show that early SCPs and late migratory crest cells have similar transcriptional profiles characterised by a multipotent "hub" state containing cells biased towards traditional neural crest fates. SCPs keep diverging from the neural crest after being primed towards terminal Schwann cells and other fates, with different subtypes residing in distinct anatomical locations. Functional experiments using CRISPR-Cas9 loss-of-function further show that knockout of the common "hub" gene Sox8 causes defects in neural crest-derived cells along peripheral nerves by facilitating differentiation of SCPs towards sympathoadrenal fates. Finally, specific tumour populations found in melanoma, neurofibroma and neuroblastoma map to different stages of SCP/Schwann cell development. Overall, SCPs resemble migrating neural crest cells that maintain multipotency and become transcriptionally primed towards distinct lineages.
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
