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.
- MeSH
- Cell Differentiation physiology MeSH
- Adult MeSH
- Genetic Vectors genetics MeSH
- Induced Pluripotent Stem Cells * physiology transplantation MeSH
- Rats MeSH
- Humans MeSH
- Spinal Cord MeSH
- Brain MeSH
- Neural Stem Cells * physiology transplantation MeSH
- Specimen Handling methods MeSH
- Tissue and Organ Harvesting methods MeSH
- Swine MeSH
- Cellular Reprogramming * genetics physiology MeSH
- Graft Survival physiology MeSH
- Injections, Spinal * adverse effects instrumentation methods MeSH
- Stem Cell Transplantation * adverse effects instrumentation methods MeSH
- Sendai virus MeSH
- Treatment Outcome MeSH
- Animals MeSH
- Check Tag
- Adult MeSH
- Rats MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The critical requirements in developing clinical-grade human-induced pluripotent stem cells-derived neural precursors (hiPSCs-NPCs) are defined by expandability, genetic stability, predictable in vivo post-grafting differentiation, and acceptable safety profile. Here, we report on the use of manual-selection protocol for generating expandable and stable human NPCs from induced pluripotent stem cells. The hiPSCs were generated by the reprogramming of peripheral blood mononuclear cells with Sendai-virus (SeV) vector encoding Yamanaka factors. After induction of neural rosettes, morphologically defined NPC colonies were manually harvested, re-plated, and expanded for up to 20 passages. Established NPCs showed normal karyotype, expression of typical NPCs markers at the proliferative stage, and ability to generate functional, calcium oscillating GABAergic or glutamatergic neurons after in vitro differentiation. Grafted NPCs into the striatum or spinal cord of immunodeficient rats showed progressive maturation and expression of early and late human-specific neuronal and glial markers at 2 or 6 months post-grafting. No tumor formation was seen in NPCs-grafted brain or spinal cord samples. These data demonstrate the effective use of in vitro manual-selection protocol to generate safe and expandable NPCs from hiPSCs cells. This protocol has the potential to be used to generate GMP (Good Manufacturing Practice)-grade NPCs from hiPSCs for future clinical use.
- MeSH
- Cell Differentiation MeSH
- Induced Pluripotent Stem Cells * MeSH
- Rats MeSH
- Leukocytes, Mononuclear MeSH
- Humans MeSH
- Neural Stem Cells * MeSH
- Neurons metabolism MeSH
- Sendai virus genetics MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Only very few comparative studies have been performed that evaluate general trends of virus growth under 3D in comparison with 2D cell culture conditions. The aim of this study was to investigate differences when four animal viruses are cultured in 2D and 3D. Suid herpesvirus 1 (SuHV-1), Vesicular stomatitis virus (VSIV), Bovine adenovirus (BAdV) and Bovine parainfluenza 3 virus (BPIV-3) were cultivated in 3D rotating wall vessels (RWVs) and conventional 2D cultures. The production of virus particles, the portion of infectious particles, and the infectious growth curves were compared. For all viruses, the production of virus particles (related to cell density), including the non-infectious ones, was lower in 3D than in 2D culture. The production of only infectious particles was significantly lower in BAdV and BPIV-3 in 3D cultures in relation to cell density. The two cultivation approaches resulted in significantly different virus particle-to-TCID50 ratios in three of the four viruses: lower in SuHV-1 and BPIV-3 and higher in BAdV in 3D culture. The infectious virus growth rates were not significantly different in all viruses. Although 3D RWV culture resulted in lower production of virus particles compared to 2D systems, the portion of infectious particles was higher for some viruses.
- MeSH
- Atadenovirus growth & development physiology ultrastructure MeSH
- Cell Culture Techniques * instrumentation methods MeSH
- Madin Darby Canine Kidney Cells MeSH
- Chlorocebus aethiops MeSH
- Virus Cultivation methods MeSH
- Swine MeSH
- Herpesvirus 1, Suid growth & development physiology ultrastructure MeSH
- Dogs MeSH
- Virus Replication MeSH
- Cattle MeSH
- Vero Cells MeSH
- Parainfluenza Virus 3, Bovine growth & development physiology ultrastructure MeSH
- Vesicular stomatitis Indiana virus growth & development physiology ultrastructure MeSH
- Animals MeSH
- Check Tag
- Dogs MeSH
- Cattle MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
Prevalence opakujících se epizod průduškové obstrukce je u kojenců a batolat vysoká. Virové infekce dýchacích cest, zejména respirační syncytiální virrus (RSV) a lidské rinoviry (HRV), vyvolávají nejčastěji pískoty u dětí v prvních letech života. Z klinického hlediska se u dětí mladších 5 let rozlišují občasné pískoty při dýchání, které jsou výlučně vyvolány virovou infekcí, a pískoty způsobené více faktory. V současnosti je imunoprofylaxe RSV infekcí palivizumabem u části pacientů prevencí vzniku pískotů.
Prevalence of reccurent wheeze in infants and toddlers is high. Viral respiratory infections, particularly with respiratory syncytial virus (RSV) and human rhinovirus (HRV), are the most common causes of wheezing. For clinical purposes, reccurent wheeze in children below the age of 5 years is classified as episodic (viral) or multiple-trigger wheeze. Currently, immunoprophylaxis with palivizumab RSV infections is the prevention of wheezing in some patients.
- Keywords
- imunoprofylaxe,
- MeSH
- Antiviral Agents therapeutic use MeSH
- Asthma * diagnosis complications physiopathology MeSH
- Diagnosis, Differential MeSH
- Child MeSH
- Antibodies, Monoclonal, Humanized therapeutic use MeSH
- Immunization methods MeSH
- Respiratory Tract Infections etiology immunology prevention & control MeSH
- Respiratory Syncytial Virus Infections prevention & control MeSH
- Respirovirus Infections prevention & control MeSH
- Humans MeSH
- Disease Susceptibility MeSH
- Palivizumab MeSH
- Recurrence MeSH
- Respiratory Syncytial Viruses immunology MeSH
- Respiratory Sounds * diagnosis immunology drug effects MeSH
- Respirovirus immunology MeSH
- Rhinovirus genetics immunology MeSH
- Risk Factors MeSH
- Virus Diseases * etiology complications MeSH
- Check Tag
- Child MeSH
- Humans MeSH
- Publication type
- Review MeSH
- MeSH
- Incidence MeSH
- Respiratory Tract Infections epidemiology microbiology MeSH
- Humans MeSH
- Mycoplasma pneumoniae isolation & purification MeSH
- Orthomyxoviridae isolation & purification MeSH
- Respiratory Syncytial Viruses isolation & purification MeSH
- Respirovirus isolation & purification MeSH
- Check Tag
- Humans MeSH
- Geographicals
- Czech Republic MeSH
- MeSH
- Child MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Fluoroimmunoassay MeSH
- Paramyxoviridae Infections diagnosis immunology MeSH
- Humans MeSH
- Parainfluenza Virus 1, Human isolation & purification MeSH
- Parainfluenza Virus 2, Human MeSH
- Serologic Tests MeSH
- Hemagglutination Inhibition Tests MeSH
- Check Tag
- Child MeSH
- Humans MeSH
Acta hygienica, epidemiologica et microbiologica, ISSN 0862-5956 Příl. č. 15/ 1991
11 s. : il., tab. ; 21 cm
- MeSH
- Antigens, Viral analysis MeSH
- Enzyme-Linked Immunosorbent Assay methods MeSH
- Immunologic Techniques methods standards MeSH
- Paramyxoviridae Infections diagnosis MeSH
- Parainfluenza Virus 1, Human MeSH
- Parainfluenza Virus 2, Human MeSH
- Parainfluenza Virus 3, Human MeSH
- Publication type
- Guideline MeSH
- Conspectus
- Patologie. Klinická medicína
- NML Fields
- chemie, klinická chemie
- virologie
In 1981 we examined 247 sera for the presence of antibodies against all three types of poliovirus and 253 sera for antibodies against M. parotitidis and three types of M. parainfluenzae viruses. The sera were obtained from the cord blood of mothers between 15 and 34 years of age. All mothers were divided into four age groups, each with primipara and multipara subgroups. The rate of seropositivity for type 1 and type 2 poliovirus-specific antibody was in all age groups higher than 90%, the overall seropositivity rate for type 3 poliovirus antibody was 83.6%, with 73.1% as the lowest rate for age group of youngest mothers. Significant seropositivity variations between the primipara and multipara subgroups were recorded only for type 3 antibody in the two age groups of oldest mothers (25-29 and 30-34 years). This is consistent with the assumed booster effect of Sabin vaccine strains on mothers of families with more than one child. Antibodies specific to mumps virus were present in the cord blood of 78.6% of all mothers and the rates of seropositivity were found to rise with the increasing age. Seropositivity for M. parainfluenzae type 1-specific antibody was demonstrated in 95.8%, for type 2-specific antibody in 98.9% and for type 3-specific antibody in 100% of mothers, which is suggestive of high herd immunity levels in the population.
- MeSH
- Adult MeSH
- Fetal Blood immunology MeSH
- Paramyxoviridae Infections immunology MeSH
- Humans MeSH
- Maternal-Fetal Exchange MeSH
- Adolescent MeSH
- Placenta immunology MeSH
- Poliomyelitis immunology MeSH
- Poliovirus Vaccine, Oral immunology MeSH
- Poliovirus immunology MeSH
- Mumps immunology MeSH
- Antibodies, Viral immunology MeSH
- Respirovirus immunology MeSH
- Pregnancy MeSH
- Age Factors MeSH
- Mumps virus immunology MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Adolescent MeSH
- Pregnancy MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
