Despite constant advances in the field of pediatric oncology, the survival rate of high-risk neuroblastoma patients remains poor. The molecular and genetic features of neuroblastoma, such as MYCN amplification and stemness status, have established themselves not only as potent prognostic and predictive factors but also as intriguing targets for personalized therapy. Novel thiosemicarbazones target both total level and activity of a number of proteins involved in some of the most important signaling pathways in neuroblastoma. In this study, we found that di-2-pyridylketone 4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC) potently decreases N-MYC in MYCN-amplified and c-MYC in MYCN-nonamplified neuroblastoma cell lines. Furthermore, DpC succeeded in downregulating total EGFR and phosphorylation of its most prominent tyrosine residues through the involvement of NDRG1, a positive prognostic marker in neuroblastoma, which was markedly upregulated after thiosemicarbazone treatment. These findings could provide useful knowledge for the treatment of MYC-driven neuroblastomas that are unresponsive to conventional therapies.

• MeSH
• amplifikace genu účinky léků   MeSH
• biologické modely MeSH
• chelátory železa farmakologie   MeSH
• down regulace účinky léků   MeSH
• erbB receptory metabolismus   MeSH
• fosforylace účinky léků   MeSH
• fyziologický stres účinky léků   MeSH
• intracelulární signální peptidy a proteiny metabolismus   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• neuroblastom metabolismus   patologie   MeSH
• proteiny buněčného cyklu metabolismus   MeSH
• protoonkogen n-myc metabolismus   MeSH
• protoonkogenní proteiny c-akt metabolismus   MeSH
• pyridiny farmakologie   MeSH
• signální transdukce * MeSH
• thiosemikarbazony farmakologie   MeSH
• tvar buňky účinky léků   MeSH
• umlčování genů účinky léků   MeSH
• upregulace účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Diamond-like carbon (DLC) is a biocompatible material that has many potential biomedical applications, including in orthopaedics. DLC layers doped with Cr at atomic percent (at.%) of 0, 0.9, 1.8, 7.3, and 7.7 at.% were evaluated with reference to their osteoinductivity with human bone marrow mesenchymal stromal cells (hMSCs), immune activation potential with RAW 264.7 macrophage-like cells, and their effect on apoptosis in Saos-2 human osteoblast-like cells and neonatal human dermal fibroblasts (NHDFs). At mRNA level, hMSCs on DLC doped with 0.9 and 7.7 at.% of Cr reached higher maximum values of both RUNX2 and alkaline phosphatase. An earlier onset of mRNA production of type I collagen and osteocalcin was also observed on these samples; they also supported the production of both type I collagen and osteocalcin. RAW 264.7 macrophages were screened using a RayBio™ Human Cytokine Array for cytokine production. 10 cytokines were at a concentration more than 2 × as high as the concentration of a positive control, but the values for the DLC samples were only moderately higher than the values on glass. NHDF cells, but not Saos-2 cells, had a higher expression of pro-apoptotic markers Bax and Bim and a lower expression of anti-apoptotic factor BCL-XL in proportion to the Cr content. Increased apoptosis was also proven by annexin V staining. These results show that a Cr-doped DLC layer with a lower Cr content can act as an osteoinductive material with relatively low immunogenicity, but that a higher Cr content can induce cell apoptosis.

• MeSH
• aktiny metabolismus   MeSH
• alkalická fosfatasa genetika   metabolismus   MeSH
• apoptóza účinky léků   imunologie   MeSH
• buněčná adheze účinky léků   MeSH
• buněčná diferenciace účinky léků   imunologie   MeSH
• buněčné linie MeSH
• chrom farmakologie   MeSH
• cytokiny metabolismus   MeSH
• diamant farmakologie   MeSH
• fibroblasty cytologie   účinky léků   MeSH
• kolagen typu I genetika   metabolismus   MeSH
• lidé MeSH
• makrofágy účinky léků   metabolismus   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   imunologie   metabolismus   MeSH
• myši MeSH
• osteogeneze účinky léků   MeSH
• osteokalcin genetika   metabolismus   MeSH
• počet buněk MeSH
• proliferace buněk účinky léků   MeSH
• protein PEBP2alfaA genetika   metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• RNA metabolismus   MeSH
• tvar buňky účinky léků   MeSH
• vápník metabolismus   MeSH
• vinkulin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In vitro produced β-like cells can provide promising cell therapy for curing the epidemic of diabetes. In this context, we aimed to investigate the effects of different concentrations of γ-aminobutyric acid (GABA) on the differentiation of rat pancreatic ductal epithelial-like stem cells (PDESCs) into β-like cells. The PDESC line cells were cultured in the basal media (DMEM/F12 + 10% FBS + 1% penicillinstreptomycin) supplemented with 0 μM, 5 μM, 50 μM, 500 μM, and 5 mM of GABA for 28 days to induce their differentiation. The differentiated cells were detected by cell morphology, dithizone (DTZ) staining, immunofluorescence staining, real-time polymerase chain reaction (qPCR), and glucose-stimulated insulin secretion (GSIS) assay to validate their identity. At the end of 28 days, compared with the control group, enrichment of induced cells was high among the 5 μM, 50 μM, 500 μM, and 5 mM GABA induction groups. The formation of islet-like cell clusters (ICCs) began at 14 days, and the cell clusters showed a growth trend with the culture time. The induced ICCs were positive for DTZ staining, while the control group showed negative results for DTZ staining and the differentiated cells were also positive for β-cell-specific markers (Ins1 and Pdx1). GSIS assay of 50 μM induction group cells at 28 days showed significantly higher levels of C-peptide and insulin secretion than the control, 5 μM, 500 μM, and 5 mM GABA-treated groups (P < 0.01). At the same time, the 50 μM induction group cells also showed significantly higher levels of Ins1, Pdx1 and Nkx6.1 mRNA as compared to the 5 μM, 500 μM and 5 mM GABA groups (P < 0.01). Thus, the addition of GABA to the basal medium effectively induced differentiation of adult rat PDESCs into insulin-secreting β-like cells, and 50 μM was the most effective concentration for the induction.

• MeSH
• agregace buněk účinky léků   MeSH
• beta-buňky cytologie   metabolismus   MeSH
• buněčná diferenciace účinky léků   MeSH
• C-peptid metabolismus   MeSH
• GABA farmakologie   MeSH
• homeodoménové proteiny genetika   metabolismus   MeSH
• inzulin metabolismus   MeSH
• kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• potkani Sprague-Dawley MeSH
• regulace genové exprese účinky léků   MeSH
• trans-aktivátory genetika   metabolismus   MeSH
• tvar buňky účinky léků   MeSH
• vývody pankreatu cytologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Neural rosette formation is a critical morphogenetic process during neural development, whereby neural stem cells are enclosed in rosette niches to equipoise proliferation and differentiation. How neural rosettes form and provide a regulatory micro-environment remains to be elucidated. We employed the human embryonic stem cell-based neural rosette system to investigate the structural development and function of neural rosettes. Our study shows that neural rosette formation consists of five types of morphological change: intercalation, constriction, polarization, elongation and lumen formation. Ca2+ signaling plays a pivotal role in the five steps by regulating the actions of the cytoskeletal complexes, actin, myosin II and tubulin during intercalation, constriction and elongation. These, in turn, control the polarizing elements, ZO-1, PARD3 and β-catenin during polarization and lumen production for neural rosette formation. We further demonstrate that the dismantlement of neural rosettes, mediated by the destruction of cytoskeletal elements, promotes neurogenesis and astrogenesis prematurely, indicating that an intact rosette structure is essential for orderly neural development.

• MeSH
• aktiny metabolismus   MeSH
• apoptóza účinky léků   MeSH
• buněčný rodokmen účinky léků   MeSH
• cytoskelet účinky léků   metabolismus   MeSH
• lidé MeSH
• lidské embryonální kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• myosin typu II metabolismus   MeSH
• nervové kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• neurogeneze účinky léků   MeSH
• neurony cytologie   účinky léků   metabolismus   ultrastruktura   MeSH
• polarita buněk účinky léků   MeSH
• protein zonula occludens 1 metabolismus   MeSH
• tvar buňky * účinky léků   MeSH
• tvorba rozet * MeSH
• vápník farmakologie   MeSH
• vápníková signalizace * účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Polymeric biomaterials are widely used in medical applications owing to their low cost, processability and sufficient toughness. Surface modification by creating a thin film of bioactive agents is promising technique to enhance cellular interactions, regulate the protein adsorption and/or avoid bacterial infections. Polyethylene is one of the most used polymeric biomaterial but its hydrophobic nature impedes its further chemical modifications. Plasma treatment is unique method to increase its hydrophilicity by incorporating hydrophilic oxidative functional groups and tailoring the surface by physical etching. Furthermore, grafting of polymer brushes of amine group containing monomers onto the functionalized surface lead to strongly immobilized bioactive agents at the final step. Chondroitin sulphate is natural polysaccharide mainly found in connective cartilage tissue which used as a bioactive agent to immobilize onto polyethylene surface by multistep method in this study.

• MeSH
• chondroitin sulfáty chemie   farmakologie   MeSH
• fibroblasty cytologie   účinky léků   MeSH
• fotoelektronová spektroskopie MeSH
• mikroskopie atomárních sil MeSH
• myši MeSH
• proliferace buněk účinky léků   MeSH
• skot MeSH
• smáčivost MeSH
• spektroskopie infračervená s Fourierovou transformací MeSH
• tvar buňky účinky léků   MeSH
• voda chemie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Bone and cartilage are tissues of a three-dimensional (3D) nature. Therefore, scaffolds for their regeneration should support cell infiltration and growth in all 3 dimensions. To fulfill such a requirement, the materials should possess large, open pores. Centrifugal spinning is a simple method for producing 3D fibrous scaffolds with large and interconnected pores. However, the process of bone regeneration is rather complex and requires additional stimulation by active molecules. In the current study, we introduced a simple composite scaffold based on platelet adhesion to poly-ε-caprolactone 3D fibers. Platelets were used as a natural source of growth factors and cytokines active in the tissue repair process. By immobilization in the fibrous scaffolds, their bioavailability was prolonged. The biological evaluation of the proposed system in the MG-63 model showed improved metabolic activity, proliferation and alkaline phosphatase activity in comparison to nonfunctionalized fibrous scaffold. In addition, the response of cells was dose dependent with improved biocompatibility with increasing platelet concentration. The results demonstrated the suitability of the system for bone tissue.

• MeSH
• adhezivita trombocytů účinky léků   MeSH
• alkalická fosfatasa metabolismus   MeSH
• kinetika MeSH
• lidé MeSH
• mezibuněčné signální peptidy a proteiny aplikace a dávkování   farmakologie   MeSH
• nádorové buněčné linie MeSH
• osteoblasty cytologie   účinky léků   ultrastruktura   MeSH
• osteogeneze účinky léků   MeSH
• polyestery chemie   farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• systémy cílené aplikace léků metody   MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury chemie   MeSH
• trombocyty účinky léků   metabolismus   ultrastruktura   MeSH
• tvar buňky účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

This study was focused on characterizing the differentiation of bone marrow-derived mesenchymal stem cells (MSCs) into corneal-like cells. Mouse MSCs were isolated from the bone marrow, grown in cell culture for 3 weeks, and purified using a magnetic activated cell sorter. Purified MSCs were cultured with an extract prepared from excised corneas and in the presence or absence of insulin-like growth factor-I (IGF-I). Analysis by quantitative real-time polymerase chain reaction showed that the expression of corneal specific markers, such as cytokeratin 12 (K12), keratocan, and lumican, was already induced after a 3-day cultivation and gradually increased during the 10-day incubation of MSCs with the extract. The presence of IGF-I significantly increased differentiation. Immunofluorescence analysis of differentiated MSCs showed positive results for the K12 protein. The morphology of the differentiated cells and the expression of cell surface markers CD45, CD11b, CD73, CD44, and CD105 were comparable in the control and differentiated MSCs. Proliferative activity was even higher in differentiated cells than in untreated MSCs. Both untreated and differentiated MSCs inhibited the production of interleukin-2 and interferon-γ in spleen cells stimulated with Concanavalin A. The results thus show that MSCs cultured in the presence of corneal extract and IGF-I efficiently differentiate into corneal-like cells. The differentiated cells possess characteristics of corneal epithelial cells and keratocytes, while at the same time maintaining MSC properties.

• MeSH
• biologické markery metabolismus   MeSH
• buněčná diferenciace účinky léků   genetika   MeSH
• imunosupresivní léčba MeSH
• insulinu podobný růstový faktor I farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• myši inbrední BALB C MeSH
• proliferace buněk účinky léků   genetika   MeSH
• regulace genové exprese účinky léků   MeSH
• rohovka cytologie   MeSH
• tvar buňky účinky léků   genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The paper is focused on pilot study of effects of novel synthetic protein kinase inhibitors-maleimide derivatives in different concentrations on normal, transformed and multipotent cell lines. Influence on cell proliferation and morphological characteristics has been demonstrated. The chosen agents cause antiproliferative effect on transformed cells and are not cytotoxic to normal cell lines. Moreover, different maleimide derivatives' effects on multipotent cells in attached and floating states has been shown. Described results can be used for further research of the maleimide derivatives as antitumor agents.

• MeSH
• buňky cytologie   účinky léků   MeSH
• endoteliální buňky cytologie   účinky léků   MeSH
• fibroblasty cytologie   účinky léků   MeSH
• HEK293 buňky MeSH
• HeLa buňky MeSH
• inhibiční koncentrace 50 MeSH
• kultivované buňky MeSH
• lidé MeSH
• maleimidy chemie   farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   MeSH
• MFC-7 buňky MeSH
• multipotentní kmenové buňky cytologie   účinky léků   MeSH
• pupečník cytologie   MeSH
• transformované buněčné linie MeSH
• tvar buňky účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Foetal calf serum (FCS) is a standard supplement used in media for in vitro stem cell cultivation. This xenogeneic supplement remains widely used for its favourable growth-promoting properties and ease of accessibility; however, it is inherently not fit for human medicine due to its capacity to temper with the cultured cell quality. For this reason, the international community encourages research and development of allogeneic sera, which would expunge this issue. This study aims to investigate the differences in proliferative capacity, phenotype, and differentiation capacity of ecto-mesenchymal stem cells from human exfoliated deciduous teeth (SHED) cultured in vitro in media supplemented with allogeneic and xenogeneic sera. To address these aims, we cultured three lineages of stem cells in media supplemented with FCS in a concentration of 2% + growth factors; human blood plasma and platelet-rich plasma in concentrations of 2% + growth factors, and 10%. Here, the xenogeneic cultivation was considered as a basis for comparison because this serum is commonly used in studies concerning ecto-mesenchymal stem cells. The study shows that multipotent ecto-mesenchymal SHED can be feasibly cultivated in media where the xenogeneic FCS is substituted by allogeneic platelet-rich plasma, considering the cultured cell proliferative and differentiation capacities. We have also proved that different sera impact the cultured cells' phenotype differently, which has major implications for previous and future stem cell research and regenerative therapy.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• chondrogeneze účinky léků   MeSH
• dítě MeSH
• fenotyp MeSH
• kultivační média farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   MeSH
• neurogeneze účinky léků   MeSH
• osteogeneze účinky léků   MeSH
• předškolní dítě MeSH
• proliferace buněk účinky léků   MeSH
• tvar buňky účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• vývoj svalů účinky léků   MeSH
• zuby mléčné cytologie   MeSH
• zvířata MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Plant cell morphogenesis involves concerted rearrangements of microtubules and actin microfilaments. We previously reported that FH1, the main Arabidopsis thaliana housekeeping Class I membrane-anchored formin, contributes to actin dynamics and microtubule stability in rhizodermis cells. Here we examine the effects of mutations affecting FH1 (At3g25500) on cell morphogenesis and above-ground organ development in seedlings, as well as on cytoskeletal organization and dynamics, using a combination of confocal and variable angle epifluorescence microscopy with a pharmacological approach. Homozygous fh1 mutants exhibited cotyledon epinasty and had larger cotyledon pavement cells with more pronounced lobes than the wild type. The pavement cell shape alterations were enhanced by expression of the fluorescent microtubule marker GFP-microtubule-associated protein 4 (MAP4). Mutant cotyledon pavement cells exhibited reduced density and increased stability of microfilament bundles, as well as enhanced dynamics of microtubules. Analogous results were also obtained upon treatments with the formin inhibitor SMIFH2 (small molecule inhibitor of formin homology 2 domains). Pavement cell shape in wild-type (wt) and fh1 plants in some situations exhibited a differential response towards anti-cytoskeletal drugs, especially the microtubule disruptor oryzalin. Our observations indicate that FH1 participates in the control of microtubule dynamics, possibly via its effects on actin, subsequently influencing cell morphogenesis and macroscopic organ development.

• MeSH
• aktiny metabolismus   MeSH
• Arabidopsis cytologie   účinky léků   metabolismus   MeSH
• biologické markery metabolismus   MeSH
• biologické modely MeSH
• cytoskelet účinky léků   metabolismus   MeSH
• fluorescence MeSH
• klathrin metabolismus   MeSH
• kotyledon účinky léků   metabolismus   MeSH
• membránové proteiny metabolismus   MeSH
• mikrofilamenta účinky léků   metabolismus   MeSH
• mikrotubuly účinky léků   metabolismus   MeSH
• mutace genetika   MeSH
• proteiny huseníčku metabolismus   MeSH
• semenáček účinky léků   růst a vývoj   metabolismus   MeSH
• thioketony farmakologie   MeSH
• tvar buňky * účinky léků   MeSH
• uracil analogy a deriváty   farmakologie   MeSH
• zelené fluorescenční proteiny metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH