Endodermal differentiation
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The potential clinical applications of hematopoietic stem cells (HSCs) derived from human pluripotent stem cells (hPSCs) are limited by the difficulty of recapitulating embryoid hematopoiesis and by the unknown differentiation potential of hPSC lines. To evaluate their hematopoietic developmental potential, available hPSC lines were differentiated by an embryoid body (EB) suspension culture in serum-free medium supplemented with three different cytokine mixes (CMs). The hPSC differentiation status was investigated by the flow cytometry expression profiles of cell surface molecules, and the gene expression of pluripotency and differentiation markers over time was evaluated by real-time reverse transcription polymerase chain reaction (qRT-PCR). hPSC lines differed in several aspects of the differentiation process, including the absolute yield of hematopoietic progenitors, the proportion of hematopoietic progenitor populations, and the effect of various CMs. The ability to generate hematopoietic progenitors was then associated with the morphology of the developing EBs, the expression of the endodermal markers AFP and SOX17, and the hematopoietic transcription factor RUNX1. These findings deepen the knowledge about the hematopoietic propensity of hPSCs and identify its variability as an aspect that must be taken into account before the usage of hPSC-derived HSCs in downstream applications.
- MeSH
- buněčné linie MeSH
- diferenciační antigeny biosyntéza MeSH
- embryoidní tělíska cytologie metabolismus MeSH
- endoderm cytologie metabolismus MeSH
- hematopoéza * MeSH
- lidé MeSH
- lidské embryonální kmenové buňky cytologie metabolismus MeSH
- regulace genové exprese * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
During mouse preimplantation embryo development, the classically described second cell-fate decision involves the specification and segregation, in blastocyst inner cell mass (ICM), of primitive endoderm (PrE) from pluripotent epiblast (EPI). The active role of fibroblast growth factor (Fgf) signalling during PrE differentiation, particularly in the context of Erk1/2 pathway activation, is well described. However, we report that p38 family mitogen-activated protein kinases (namely p38α/Mapk14 and p38β/Mapk11; referred to as p38-Mapk14/11) also participate in PrE formation. Specifically, functional p38-Mapk14/11 are required, during early-blastocyst maturation, to assist uncommitted ICM cells, expressing both EPI and earlier PrE markers, to fully commit to PrE differentiation. Moreover, functional activation of p38-Mapk14/11 is, as reported for Erk1/2, under the control of Fgf-receptor signalling, plus active Tak1 kinase (involved in non-canonical bone morphogenetic protein (Bmp)-receptor-mediated PrE differentiation). However, we demonstrate that the critical window of p38-Mapk14/11 activation precedes the E3.75 timepoint (defined by the initiation of the classical 'salt and pepper' expression pattern of mutually exclusive EPI and PrE markers), whereas appropriate lineage maturation is still achievable when Erk1/2 activity (via Mek1/2 inhibition) is limited to a period after E3.75. We propose that active p38-Mapk14/11 act as enablers, and Erk1/2 as drivers, of PrE differentiation during ICM lineage specification and segregation.
- MeSH
- blastocysta fyziologie MeSH
- buněčná diferenciace MeSH
- embryonální vývoj * MeSH
- endoderm embryologie MeSH
- fibroblastové růstové faktory metabolismus MeSH
- messenger RNA metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 11 metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 14 metabolismus MeSH
- myši MeSH
- signální transdukce MeSH
- zárodečné listy fyziologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Prezentujeme kazuistiku 41letého pacienta, který byl přivezen do našeho zdravotnického zařízení pro podezření na krvácení do horní části gastrointestinálního traktu. Gastroskopicky byla nalezena exulcerovaná infiltrace na zadní stěně subkardiální části žaludku. Histologicky byl prokázán vzácný adenokarcinom žaludku s yolk sac diferenciací. Tento typ nádoru patří mezi malignity se špatnou prognózou. V době diagnózy bývá již přítomna generalizace s jaterním metastatickým postižením.
We present the case of a 41-year-old patient who was brought to our hospital because of suspected bleeding into the upper part of gastrointestinal tract. Gastroscopy found an ulcerated infiltration at the posterior wall of the subcardial stomach. Histological analysis showed the presence of an adenocarcinoma of the stomach with yolk sac differentiation, which is a rare type of malignant tumor with a poor prognosis that has usually metastasized to secondary sites such as the liver at the time of diagnosis.
- Klíčová slova
- protokol BEP,
- MeSH
- adenokarcinom diagnóza farmakoterapie patologie MeSH
- alfa-fetoproteiny * metabolismus MeSH
- bleomycin aplikace a dávkování MeSH
- cisplatina aplikace a dávkování MeSH
- diferenciální diagnóza MeSH
- dospělí MeSH
- etoposid aplikace a dávkování MeSH
- fatální výsledek MeSH
- imunohistochemie MeSH
- lidé MeSH
- nádor z entodermálního sinusu * diagnóza farmakoterapie patologie MeSH
- nádorové biomarkery krev MeSH
- nádory jater sekundární MeSH
- nádory žaludku * diagnóza farmakoterapie patologie MeSH
- protokoly protinádorové kombinované chemoterapie aplikace a dávkování škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
Post-translational modification of histones is fundamental to the regulation of basic nuclear processes and subsequent cellular events, including differentiation. In this study, we analyzed acetylated forms of histones H2A, H2B, and H4 during induced differentiation in mouse (mESCs) and human (hESCs) embryonic stem cells and during induced enterocytic differentiation of colon cancer cells in vitro. Endoderm-like differentiation of mESCs induced by retinoic acid and enterocytic differentiation induced by histone deacetylase inhibitor sodium butyrate were accompanied by increased mono-, di-, and tri-acetylation of histone H2B and a pronounced increase in di- and tri-acetylation of histone H4. In enterocytes, mono-acetylation of histone H2A also increased and tetra-acetylation of histone H4 appeared only after induction of this differentiation pathway. During differentiation of hESCs, we observed increased mono-acetylation and decreased tri-acetylation of H2B. Mono-, di-, and tri-acetylation of H4 were reduced, manifested by a significant increase in nonacetylated H4 histones. Levels of acetylated histones increased during induced differentiation in mESCs and during histone deacetylase (HDAC) inhibitor-induced enterocytic differentiation, whereas differentiation of human ESCs was associated with reduced acetylation of histones H2B and H4.
- MeSH
- acetylace MeSH
- buněčná diferenciace MeSH
- embryonální kmenové buňky cytologie metabolismus MeSH
- endoderm cytologie metabolismus MeSH
- enterocyty cytologie metabolismus MeSH
- epigeneze genetická MeSH
- histonacetyltransferasy metabolismus MeSH
- histony metabolismus MeSH
- lidé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- posttranslační úpravy proteinů * MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Epigenetic marks are important factors regulating the pluripotency and differentiation of human embryonic stem cells (hESCs). In this study, we analyzed H3K9 acetylation, an epigenetic mark associated with transcriptionally active chromatin, during endoderm-like differentiation of hESCs. ChIP-on-chip analysis revealed that differentiation results in a genome-wide decrease in promoter H3K9 acetylation. Among the 24,659 promoters analyzed, only 117 are likely to be involved in pluripotency, while 25 acetylated promoters are likely to be responsible for endoderm-like differentiation. In pluripotent hESCs, the chromosomes with the highest absolute levels of H3K9 acetylation are chromosomes 1, 6, 2, 17, 11, and 12 (listed in order of decreasing acetylation). Chromosomes 17, 19, 11, 20, 22, and 12 are the most prone to differentiation-related changes (both increased acetylation and deacetylation). When chromosome size (in Mb) was accounted for, the highest H3K9 acetylation levels were found on chromosome 19, 17, 6, 12, 11, and 1, and the greatest differentiation-associated decreases in H3K9 acetylation occurred on chromosomes 19, 17, 11, 12, 16, and 1. The gene density and size of individual chromosomes were strongly correlated with the levels of H3K9 acetylation. Our analyses point to chromosomes 11, 12, 17, and 19 as being critical for hESC pluripotency and endoderm-like differentiation. J. Cell. Physiol. 219: 677-687, 2009. (c) 2009 Wiley-Liss, Inc.
- MeSH
- acetylace MeSH
- buněčná diferenciace fyziologie genetika MeSH
- buněčné linie MeSH
- chromatinová imunoprecipitace MeSH
- embryonální kmenové buňky cytologie metabolismus MeSH
- epigeneze genetická MeSH
- financování organizované MeSH
- genom lidský MeSH
- histony genetika chemie metabolismus MeSH
- lidé MeSH
- lidské chromozomy genetika metabolismus MeSH
- pluripotentní kmenové buňky cytologie metabolismus MeSH
- promotorové oblasti (genetika) MeSH
- Check Tag
- lidé MeSH
Over the past decades, the in vitro use of pluripotent cell lines gained a crucial role in toxicology, preclinical drug testing and developmental biology. NTERA2 clone D1 cells were identified as pluripotent cells with high potential for neural differentiation. Although they are commonly used cellular sources in neuropharmacology and neurodevelopmental studies, their endodermal and mesodermal differentiation potential awaits further characterization. Here, we devised improved protocols for hepatogenic and osteogenic differentiation of NTERA2 clone D1 cells. Our in vitro differentiation assays showed significant up-regulation of multiple hepatogenic markers. We also observed robust mineralization and osteogenic marker expression of NTERA2 clone D1 cells upon in vitro osteogenic induction. These results suggest that NTERA2 clone D1 cells may be utilized as an in vitro model system to study various aspects of liver biology and osteogenesis. In addition, tri-lineage differentiation of NTERA2 clone D1 cells may serve as a simple experimental control system when validating pluripotency of other cell types.
- MeSH
- buněčná diferenciace MeSH
- buněčné klony MeSH
- buněčné linie MeSH
- játra * MeSH
- osteogeneze * MeSH
- Publikační typ
- časopisecké články MeSH
