Duchenne muscular dystrophy (DMD) affects 1:3500-5000 newborn boys and manifests with progressive skeletal muscle wasting, respiratory failure and eventual heart failure. Symptoms show different onset from patients' childhood to the second decade of age. We reprogrammed fibroblasts from two independent DMD patients with a complete loss of dystrophin expression, carrying deletions of exons 45-50 and 48-50. The resulting hiPSCs show expression of pluripotency markers (NANOG, OCT4, SSEA4), differentiation capacity into all three germ layers, normal karyotype, genetic identity to the originating parental fibroblasts and the patient-specific dystrophin mutation.
- MeSH
- buněčná diferenciace MeSH
- buněčné linie cytologie metabolismus MeSH
- dítě MeSH
- Duchennova muskulární dystrofie genetika metabolismus patofyziologie MeSH
- dystrofin genetika metabolismus MeSH
- exony MeSH
- indukované pluripotentní kmenové buňky cytologie metabolismus MeSH
- lidé MeSH
- mladiství MeSH
- oktamerní transkripční faktor 3 genetika metabolismus MeSH
- sekvenční delece MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dystrofin MeSH
- oktamerní transkripční faktor 3 MeSH
BACKGROUND: The availability of tick in vitro cell culture systems has facilitated many aspects of tick research, including proteomics. However, certain cell lines have shown a tissue-specific response to infection. Thus, a more thorough characterization of tick cell lines is necessary. Proteomic comparative studies of various tick cell lines will contribute to more efficient application of tick cell lines as model systems for investigation of host-vector-pathogen interactions. RESULTS: Three cell lines obtained from a hard tick, Ixodes ricinus, and two from I. scapularis were investigated. A cell mass spectrometry approach (MALDI-TOF MS) was applied, as well as classical proteomic workflows. Using PCA, tick cell line MS profiles were grouped into three clusters comprising IRE/CTVM19 and ISE18, IRE11 and IRE/CTVM20, and ISE6 cell lines. Two other approaches confirmed the results of PCA: in-solution digestion followed by nanoLC-ESI-Q-TOF MS/MS and 2D electrophoresis. The comparison of MS spectra of the cell lines and I. ricinus tick organs revealed 29 shared peaks. Of these, five were specific for ovaries, three each for gut and salivary glands, and one for Malpighian tubules. For the first time, characteristic peaks in MS profiles of tick cell lines were assigned to proteins identified in acidic extracts of corresponding cell lines. CONCLUSIONS: Several organ-specific MS signals were revealed in the profiles of tick cell lines.
- Klíčová slova
- Biotyping, MALDI-TOF MS, Tick, Tick cell line, Tick organs,
- MeSH
- 2D gelová elektroforéza MeSH
- buněčné linie * cytologie metabolismus MeSH
- hmyzí proteiny metabolismus MeSH
- klíště cytologie MeSH
- proteomika MeSH
- slinné žlázy cytologie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice * MeSH
- tandemová hmotnostní spektrometrie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- hmyzí proteiny MeSH
Using a recombinant vaccinia virus expressing protooncogene Bcl-2, we demonstrate opposite effects of the expressed Bcl-2 in two cell lines: apoptosis induction in BSC-40 cells and apoptosis prevention in HeLa G cells. The apparent molecular weight of the expressed Bcl-2, its amounts and its effects on the mitochondrial membrane potential are comparable in both cell lines, suggesting that the consequences of Bcl-2 expression depend on the cellular environment. To further support these findings we demonstrate the pro-apoptotic effect of the expressed Bcl-2 in several other cell lines.
- MeSH
- apoptóza genetika MeSH
- buněčné linie cytologie MeSH
- Cercopithecus aethiops MeSH
- chloramfenikol-O-acetyltransferasa genetika MeSH
- druhová specificita MeSH
- epitelové buňky cytologie MeSH
- genetické vektory genetika MeSH
- geny bcl-2 * MeSH
- HeLa buňky cytologie MeSH
- intracelulární membrány fyziologie MeSH
- Jurkat buňky cytologie MeSH
- kaspasy metabolismus MeSH
- lidé MeSH
- membránové potenciály MeSH
- mitochondrie fyziologie MeSH
- protoonkogenní proteiny c-bcl-2 fyziologie MeSH
- rekombinantní fúzní proteiny fyziologie MeSH
- reportérové geny MeSH
- transfekce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- chloramfenikol-O-acetyltransferasa MeSH
- kaspasy MeSH
- protoonkogenní proteiny c-bcl-2 MeSH
- rekombinantní fúzní proteiny MeSH
- MeSH
- buněčné jádro fyziologie ultrastruktura MeSH
- buněčné linie cytologie MeSH
- lidé MeSH
- mitóza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
