For the treatment of bilateral limbal stem cell deficiency (LSCD), cell therapy with transplantation of cultivated oral mucosa epithelial cells (COMET) is a promising alternative. Although not yet established, current protocols on the cultivation of oral mucosal epithelial cell (OMECs) sheets are based mainly on substrates and xenobiotic additives that may lead to variable outcomes and undesirable immune responses by the patient. The aim of this study was to characterize OMECs cultivated in xenobiotic-free media (XF) seeded on fibrin gel, in comparison to conventional complex (COM) medium. Oral mucosal biopsies were retrieved from 31 donors. After cultivation in COM or XF medium, OMECs were compared based on growth kinetics, morphology, cell size and viability. Using immunofluorescence and gene expression analyses, the degree of stemness, proliferation and differentiation was evaluated in OMEC cultures. Our findings showed that although OMECs showed a similar morphology and viability, and comparable growth kinetics, immunofluorescence revealed the preservation of stemness (p63 + p40 positivity in cells ≤11 μm) and proliferation in both COM and XF. Gene expression analyses showed that keratin (K)13 and K15 expression levels were significantly higher in XF (adj. p < 0.001), but otherwise COM and XF-treated OMECs had comparable transcriptional profiles in a panel of stemness, proliferation and differentiation genes. These results demonstrate the feasibility of culturing OMECs on fibrin gel without xenogeneic additives, while maintaining their undifferentiated state and preserving stemness. In conclusion, both in terms of results and methodology, the procedures presented here are suitable for implementation in clinical practice.

• Klíčová slova
• Advanced therapy medicinal products, Cell culture, Fibrin glue substrate, Limbal stem cell deficiency, Oral mucosal epithelial cells, Stemness,
• MeSH
• buněčná diferenciace MeSH
• buněčné kultury * MeSH
• deficit limbálních kmenových buněk MeSH
• dospělí MeSH
• epitelové buňky * metabolismus   účinky léků   MeSH
• fibrin * MeSH
• gely MeSH
• kmenové buňky * metabolismus   cytologie   MeSH
• kultivační média MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• limbus corneae * cytologie   patologie   metabolismus   MeSH
• nemoci rohovky patologie   farmakoterapie   metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• rohovkový epitel metabolismus   cytologie   účinky léků   patologie   MeSH
• senioři MeSH
• transplantace kmenových buněk metody   MeSH
• ústní sliznice * cytologie   MeSH
• viabilita buněk MeSH
• xenobiotika farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fibrin * MeSH
• gely MeSH
• kultivační média MeSH
• xenobiotika MeSH

Dental pulp is a valuable and accessible source of stem cells (DPSCs) with characteristics similar to mesenchymal stem cells. DPSCs can regenerate a range of tissues and their potential for clinical application in regenerative medicine is promising. DPSCs have been found to express low levels of Class II HLA-DR (MHC) molecules, making them potential candidates for allogeneic transplantation without matching the donor's tissue. Research on the correlation between non-coding RNAs (ncRNAs) and human dental pulp stem cells (DPSCs) provides promising insights into the use of these cells in clinical settings for a wide range of medical conditions. It is possible to use a number of ncRNAs in order to restore the functional role of downregulated ncRNAs that are correlated with osteoblastogenesis, or to suppress the functional role of overexpressed ncRNAs associated with osteoclast differentiation in some cases.

• Klíčová slova
• Dental pulp, Non-coding RNAs, Signaling pathways, Stem cells,
• MeSH
• buněčná diferenciace MeSH
• kmenové buňky * cytologie   metabolismus   MeSH
• lidé MeSH
• nekódující RNA genetika   MeSH
• regenerativní lékařství * metody   MeSH
• transplantace kmenových buněk metody   MeSH
• zubní dřeň * cytologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• nekódující RNA MeSH

One of the challenges in clinical translation of cell-replacement therapies is the definition of optimal cell generation and storage/recovery protocols which would permit a rapid preparation of cell-treatment products for patient administration. Besides, the availability of injection devices that are simple to use is critical for potential future dissemination of any spinally targeted cell-replacement therapy into general medical practice. Here, we compared the engraftment properties of established human-induced pluripotent stem cells (hiPSCs)-derived neural precursor cell (NPCs) line once cells were harvested fresh from the cell culture or previously frozen and then grafted into striata or spinal cord of the immunodeficient rat. A newly developed human spinal injection device equipped with a spinal cord pulsation-cancelation magnetic needle was also tested for its safety in an adult immunosuppressed pig. Previously frozen NPCs showed similar post-grafting survival and differentiation profile as was seen for freshly harvested cells. Testing of human injection device showed acceptable safety with no detectable surgical procedure or spinal NPCs injection-related side effects.

• Klíčová slova
• cryopreservation, human induced pluripotent stem cells (hiPSCs), human injection device, immunosuppressed adult pig, neural precursor cells (NPCs), spinal cord,
• MeSH
• buněčná diferenciace fyziologie   MeSH
• dospělí MeSH
• genetické vektory genetika   MeSH
• indukované pluripotentní kmenové buňky * fyziologie   transplantace   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• mícha MeSH
• mozek MeSH
• nervové kmenové buňky * fyziologie   transplantace   MeSH
• odběr biologického vzorku metody   MeSH
• odběr tkání a orgánů metody   MeSH
• prasata MeSH
• přeprogramování buněk * genetika   fyziologie   MeSH
• přežívání štěpu fyziologie   MeSH
• spinální injekce * škodlivé účinky   přístrojové vybavení   metody   MeSH
• transplantace kmenových buněk * škodlivé účinky   přístrojové vybavení   metody   MeSH
• virus Sendai MeSH
• výsledek terapie MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Epilepsy is a complex disorder affecting the central nervous system and is characterised by spontaneously recurring seizures (SRSs). Epileptic patients undergo symptomatic pharmacological treatments, however, in 30% of cases, they are ineffective, mostly in patients with temporal lobe epilepsy. Therefore, there is a need for developing novel treatment strategies. Transplantation of cells releasing γ-aminobutyric acid (GABA) could be used to counteract the imbalance between excitation and inhibition within epileptic neuronal networks. We generated GABAergic interneuron precursors from human embryonic stem cells (hESCs) and grafted them in the hippocampi of rats developing chronic SRSs after kainic acid-induced status epilepticus. Using whole-cell patch-clamp recordings, we characterised the maturation of the grafted cells into functional GABAergic interneurons in the host brain, and we confirmed the presence of functional inhibitory synaptic connections from grafted cells onto the host neurons. Moreover, optogenetic stimulation of grafted hESC-derived interneurons reduced the rate of epileptiform discharges in vitro. We also observed decreased SRS frequency and total time spent in SRSs in these animals in vivo as compared to non-grafted controls. These data represent a proof-of-concept that hESC-derived GABAergic neurons can exert a therapeutic effect on epileptic animals presumably through establishing inhibitory synapses with host neurons.

• Klíčová slova
• GABA, cell integration, epilepsy, human embryonic stem cells, interneurons, optogenetics, synaptic integration,
• MeSH
• GABA metabolismus   MeSH
• hipokampus metabolismus   patologie   MeSH
• interneurony cytologie   metabolismus   MeSH
• kmenové buňky cytologie   metabolismus   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• kyselina kainová škodlivé účinky   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• recidiva MeSH
• status epilepticus chemicky indukované   metabolismus   patologie   terapie   MeSH
• transplantace kmenových buněk metody   MeSH
• záchvaty chemicky indukované   metabolismus   patologie   terapie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• GABA MeSH
• kyselina kainová 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).

• Klíčová slova
• ALS, iPS, motoneuron death, neurodegeneration, plasticity, proteoglycans, stem cells, transplantation,
• 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
• Názvy látek
• neurotrofní faktory MeSH
• proteiny regulující apoptózu MeSH
• tenascin MeSH
• versikany MeSH

Spinal cord injury (SCI) is a serious trauma, which often results in a permanent loss of motor and sensory functions, pain and spasticity. Despite extensive research, there is currently no available therapy that would restore the lost functions after SCI in human patients. Advanced treatments use regenerative medicine or its combination with various interdisciplinary approaches such as tissue engineering or biophysical methods. This review summarizes and critically discusses the research from specific interdisciplinary fields in SCI treatment such as the development of biomaterials as scaffolds for tissue repair, and using a magnetic field for targeted cell delivery. We compare the treatment effects of synthetic non-degradable methacrylate-based hydrogels and biodegradable biological scaffolds based on extracellular matrix. The systems using magnetic fields for magnetically guided delivery of stem cells loaded with magnetic nanoparticles into the lesion site are then suggested and discussed.

• Klíčová slova
• Biomaterials, Cell delivery, Hydrogel, Magnetic field, Spinal cord injury,
• MeSH
• biokompatibilní materiály farmakologie   terapeutické užití   MeSH
• hydrogely terapeutické užití   MeSH
• lidé MeSH
• magnetoterapie metody   trendy   MeSH
• poranění míchy patofyziologie   terapie   MeSH
• regenerace nervu účinky léků   fyziologie   MeSH
• transplantace kmenových buněk metody   trendy   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• biokompatibilní materiály MeSH
• hydrogely MeSH

BACKGROUND: Ciprofloxacin prophylaxis used to be a standard precaution during autologous stem cell transplantation. Its benefit, with a high prevalence of fluoroqinolone resistance in the population, has recently been under scrutiny. OBJECTIVE: To evaluate the impact of cessation of ciprofloxacin prophylaxis during stem cell transplantation for multiple myeloma. PATIENTS AND METHODS: Data from 104 patients with multiple myeloma transplanted during the period from January 2013 to April 2015 were retrospectively reviewed. 67 received standard ciprofloxacin prophylaxis (group A) and 37 received no antibacterial prophylaxis (group B). RESULTS: Febrile episodes during neutropenia, bloodstream infection (BSI) and mortality in these two cohorts were evaluated. Gram negative BSI was assessed for the colonization of quinolone-resistant gram-negative pathogens. Secondary Clostridium difficile enterocolitis presence was determined in both cohorts. There were 42 (63%), 7 (10%), and 0 febrile episodes, BSI and gram-negative BSI respectively in group A, and 34 (92%), 12 (32%), and 4 (11%) respectively in group B. The differences in the number of febrile episodes (P=0.0011) and deaths (P=0.0427) were statistically significance. Mortality was 0 and 3 (8%) in group A and group B, respectively. There was no significant difference in colonization with quinolone-resistant gram negative pathogens (25 (37%) versus 11 (30%)) between groups. The occurrence of Clostridium difficile colitis was the same in both groups. CONCLUSION: We resumed ciprofloxacin prophylaxis for the following reasons. There was a significant reduction in febrile episodes, and consequently a sparing effect of antibiotics used for treatment of this condition. No difference in Clostridium difficile colitis occurred and the mortality rate of 8% in group B was unacceptably high.

• Klíčová slova
• autologous transplantation, ciprofloxacin prophylaxis, multiple myeloma, neutropenia,
• MeSH
• antibakteriální látky terapeutické užití   MeSH
• antibiotická profylaxe * MeSH
• autologní transplantace MeSH
• bakteriální infekce prevence a kontrola   MeSH
• bakteriální léková rezistence MeSH
• ciprofloxacin terapeutické užití   MeSH
• dospělí MeSH
• gramnegativní bakteriální infekce epidemiologie   MeSH
• gramnegativní bakterie fyziologie   MeSH
• infekce vyvolané Escherichia coli epidemiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetný myelom terapie   MeSH
• přenašečství epidemiologie   MeSH
• pseudomembranózní enterokolitida epidemiologie   MeSH
• retrospektivní studie MeSH
• senioři MeSH
• stafylokokové infekce epidemiologie   MeSH
• transplantace kmenových buněk metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• ciprofloxacin MeSH

Combinatory strategies using pharmacology and stem cell therapy have emerged due to their potential in the treatment of retinal pigment epithelium (RPE) cell related diseases, and a variety of different stem cell sources have been evaluated both in animal models and in humans. RPE cells derived from human embryonic stem cells (hESCs) and human induced pluripotent cells (hiPSCs) are already in clinical trials, holding great promise for the treatment of age-related macular disease (AMD) and hereditary RPE-related retinal dystrophies. Highly efficient protocol for RPE generations have been developed, but they are still time-consuming and laborious. Areas covered: The authors review RPE related diseases, as well as the known functions of RPE cells in retinal homeostasis. The authors also discuss small molecules that target RPE in vivo as well as in vitro to aid RPE differentiation from pluripotent stem cells clinically. The authors base this review on literature searches performed through PubMed. Expert opinion: Using high-throughput systems, technology will provide the possibility of identifying and optimizing molecules/drugs that could lead to faster and simpler protocols for RPE differentiation. This could be crucial in moving forward to create safer and more efficient RPE-based personalized therapies.

• Klíčová slova
• AMD, Hipscs, RPE differentiation, cell therapy, retinal dystrophies, retinal pigment epithelial cells, small molecules,
• MeSH
• buněčná diferenciace fyziologie   MeSH
• kombinovaná terapie MeSH
• lidé MeSH
• makulární degenerace patofyziologie   terapie   MeSH
• nemoci retiny patofyziologie   terapie   MeSH
• pluripotentní kmenové buňky cytologie   MeSH
• retinální pigmentový epitel cytologie   MeSH
• rychlé screeningové testy MeSH
• transplantace kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy 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.

• Klíčová slova
• Inflammation, NF-κB, Spinal cord injury, Stem cells transplantation, TNF-α, p65,
• 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
• Názvy látek
• cytokiny MeSH
• gliový fibrilární kyselý protein MeSH
• Rela protein, rat MeSH Prohlížeč
• transkripční faktor RelA MeSH

Retinal degenerative diseases, which include age-related macular degeneration, retinitis pigmentosa, diabetic retinopathy, and glaucoma, mostly affect the elderly population and are the most common cause of decreased quality of vision or even blindness. So far, there is no satisfactory treatment protocol to prevent, stop, or cure these disorders. A great hope and promise for patients suffering from retinal diseases is represented by stem cell-based therapy that could replace diseased or missing retinal cells and support regeneration. In this respect, mesenchymal stem cells (MSCs) that can be obtained from the particular patient and used as autologous cells have turned out to be a promising stem cell type for treatment. Here we show that MSCs can differentiate into cells expressing markers of retinal cells, inhibit production of pro-inflammatory cytokines by retinal tissue, and produce a number of growth and neuroprotective factors for retinal regeneration. All of these properties make MSCs a prospective cell type for cell-based therapy of age-related retinal degenerative diseases.

• Klíčová slova
• age-related retinal degenerative diseases, mesenchymal stem cells, stem cell therapy,
• MeSH
• buněčná diferenciace genetika   fyziologie   MeSH
• degenerace retiny metabolismus   terapie   MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nemoci retiny metabolismus   terapie   MeSH
• prospektivní studie MeSH
• transplantace kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH