Nejvíce citovaný článek - PubMed ID 22873171
The key role of insulin-like growth factor I in limbal stem cell differentiation and the corneal wound-healing process
This study aimed to determine the effect of interleukin-13 (IL13) on the stemness, differentiation, proliferation, clonogenicity, and morphology of cultured limbal epithelial cells (LECs). Human limbal explants were used to culture LECs up to the second passage (P0-P2) with or without IL13 (IL13+ and IL13-, respectively). Cells were analyzed by qPCR (for the expression of ΔNp63α, BMI-1, keratin (K) 3, K7, K12, K14, K17, mucin 4, and MKI67) and immunofluorescence staining for p63α. The clonogenic ability was determined by colony-forming assay (CFA), and their metabolic activity was measured by WST-1 assay. The results of the CFA showed a significantly increased clonogenic ability in P1 and P2 cultures when LECs were cultured with IL13. In addition, the expression of putative stem cell markers (ΔNp63α, K14, and K17) was significantly higher in all IL13+ cultures compared to IL13-. Similarly, immunofluorescence analysis showed a significantly higher percentage of p63α positive cells in P2 cultures with IL13 than without it. LECs cultures without IL13 lost their cuboidal morphology with a high nucleocytoplasmic ratio after P1. The use of IL13 also led to significantly higher proliferation in P2, which can be reflected by a higher ability to reach confluence in P2 cultures. On the other hand, IL13 had no effect on corneal epithelial cell differentiation (K3 and K12 expression), and the expression of the conjunctival marker K7 significantly increased in all IL13+ cultures compared to the respective cell culture without IL13. This study showed that IL13 enhanced the stemness of LECs by increasing the clonogenicity and the expression of putative stem cell markers of LECs while maintaining their stem cell morphology. We established IL13 as a culture supplement for LESCs, which increases their stemness potential in culture, even after the second passage, and may lead to the greater success of LESCs transplantation in patients with LSCD.
- MeSH
- buněčná diferenciace MeSH
- buněčné kultury MeSH
- epitelové buňky metabolismus MeSH
- interleukin-13 metabolismus MeSH
- kmenové buňky MeSH
- kultivované buňky MeSH
- lidé MeSH
- limbus corneae * MeSH
- rohovkový epitel * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- interleukin-13 MeSH
Retinal degenerative diseases, such as age-related macular degeneration, retinitis pigmentosa, diabetic retinopathy or glaucoma, represent the main causes of a decreased quality of vision or even blindness worldwide. However, despite considerable efforts, the treatment possibilities for these disorders remain very limited. A perspective is offered by cell therapy using mesenchymal stem cells (MSCs). These cells can be obtained from the bone marrow or adipose tissue of a particular patient, expanded in vitro and used as the autologous cells. MSCs possess potent immunoregulatory properties and can inhibit a harmful inflammatory reaction in the diseased retina. By the production of numerous growth and neurotrophic factors, they support the survival and growth of retinal cells. In addition, MSCs can protect retinal cells by antiapoptotic properties and could contribute to the regeneration of the diseased retina by their ability to differentiate into various cell types, including the cells of the retina. All of these properties indicate the potential of MSCs for the therapy of diseased retinas. This view is supported by the recent results of numerous experimental studies in different preclinical models. Here we provide an overview of the therapeutic properties of MSCs, and their use in experimental models of retinal diseases and in clinical trials.
- Klíčová slova
- clinical trials, experimental models, mesenchymal stem cells, retinal degenerative diseases, stem cell therapy,
- MeSH
- autologní transplantace MeSH
- buněčná a tkáňová terapie metody MeSH
- buněčná diferenciace MeSH
- buňky kostní dřeně cytologie metabolismus MeSH
- diabetická retinopatie genetika metabolismus patologie terapie MeSH
- glaukom genetika metabolismus patologie terapie MeSH
- klinické zkoušky jako téma MeSH
- lidé MeSH
- makulární degenerace genetika metabolismus patologie terapie MeSH
- mezenchymální kmenové buňky cytologie metabolismus MeSH
- mezibuněčné signální peptidy a proteiny genetika metabolismus MeSH
- modely nemocí na zvířatech MeSH
- neurotrofní faktory genetika metabolismus MeSH
- retina metabolismus patologie MeSH
- retinopathia pigmentosa genetika metabolismus patologie terapie MeSH
- transplantace mezenchymálních kmenových buněk metody MeSH
- tuková tkáň cytologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- mezibuněčné signální peptidy a proteiny MeSH
- neurotrofní faktory MeSH
The human amniotic membrane (HAM) is widely used for its wound healing effect in clinical practice, as a feeder for the cell cultivation, or a source of cells to be used in cell therapy. The aim of this study was to find effective and safe enzymatic HAM de-epithelialization method leading to harvesting of both denuded undamaged HAM and viable human amniotic epithelial cells (hAECs). The efficiency of de-epithelialization using TrypLE Express, trypsin/ ethylenediaminetetraacetic (EDTA), and thermolysin was monitored by hematoxylin and eosin staining and by the measurement of DNA concentration. The cell viability was determined by trypan blue staining. Scanning electron microscopy and immunodetection of collagen type IV and laminin α5 chain were used to check the basement membrane integrity. De-epithelialized hAECs were cultured and their stemness properties and proliferation potential was assessed after each passage. The HAM was successfully de-epithelialized using all three types of reagents, but morphological changes in basement membrane and stroma were observed after the thermolysin application. About 60% of cells remained viable using trypsin/EDTA, approximately 6% using TrypLE Express, and all cells were lethally damaged after thermolysin application. The hAECs isolated using trypsin/EDTA were successfully cultured up to the 5th passage with increasing proliferation potential and decreased stem cell markers expression (NANOG, SOX2) in prolonged cell culture. Trypsin/EDTA technique was the most efficient for obtaining both undamaged denuded HAM and viable hAECs for consequent culture.
- MeSH
- amnion cytologie metabolismus patologie MeSH
- DNA analýza izolace a purifikace MeSH
- EDTA chemie MeSH
- epitelové buňky cytologie metabolismus patologie MeSH
- kolagen typu IV metabolismus MeSH
- kultivované buňky MeSH
- laminin metabolismus MeSH
- lidé MeSH
- mikroskopie elektronová rastrovací MeSH
- nanog metabolismus MeSH
- proliferace buněk MeSH
- reepitalizace MeSH
- transkripční faktory SOXB1 metabolismus MeSH
- trypsin metabolismus MeSH
- viabilita buněk MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA MeSH
- EDTA MeSH
- kolagen typu IV MeSH
- laminin alpha5 MeSH Prohlížeč
- laminin MeSH
- nanog MeSH
- transkripční faktory SOXB1 MeSH
- trypsin MeSH
The aim of this study was to examine the effect of molecular hydrogen (H2) on the healing of alkali-injured cornea. The effects of the solution of H2 in phosphate buffered saline (PBS) or PBS alone topically applied on the alkali-injured rabbit cornea with 0.25 M NaOH were investigated using immunohistochemical and biochemical methods. Central corneal thickness taken as an index of corneal hydration was measured with an ultrasonic pachymeter. Results show that irrigation of the damaged eyes with H2 solution immediately after the injury and then within next five days renewed corneal transparency lost after the injury and reduced corneal hydration increased after the injury to physiological levels within ten days after the injury. In contrast, in injured corneas treated with PBS, the transparency of damaged corneas remained lost and corneal hydration elevated. Later results-on day 20 after the injury-showed that in alkali-injured corneas treated with H2 solution the expression of proinflammatory cytokines, peroxynitrite, detected by nitrotyrosine residues (NT), and malondialdehyde (MDA) expressions were very low or absent compared to PBS treated injured corneas, where NT and MDA expressions were present. In conclusion, H2 solution favorably influenced corneal healing after alkali injury via suppression of oxidative stress.
- MeSH
- aktiny metabolismus MeSH
- cytokiny metabolismus MeSH
- exprese genu účinky léků MeSH
- hydroxid sodný toxicita MeSH
- imunohistochemie MeSH
- interleukin-1beta genetika metabolismus MeSH
- keratin-12 metabolismus MeSH
- keratin-3 metabolismus MeSH
- králíci MeSH
- kyselina peroxydusitá metabolismus MeSH
- malondialdehyd metabolismus MeSH
- modely nemocí na zvířatech MeSH
- oxidační stres účinky léků MeSH
- poranění rohovky etiologie metabolismus patologie MeSH
- rohovka metabolismus patologie MeSH
- vaskulární endoteliální růstový faktor A genetika metabolismus MeSH
- vodík farmakologie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- odvolaná publikace MeSH
- Názvy látek
- aktiny MeSH
- cytokiny MeSH
- hydroxid sodný MeSH
- interleukin-1beta MeSH
- keratin-12 MeSH
- keratin-3 MeSH
- kyselina peroxydusitá MeSH
- malondialdehyd MeSH
- vaskulární endoteliální růstový faktor A MeSH
- vodík MeSH
The skin-swelling test is a simple and widespread method used in field ecological research to estimate cellular immune responsiveness in animals. This immunoecological test is based on measuring the magnitude of tissue swelling response at specific times following subcutaneous application of an experimental pro-inflammatory stimulant. In the vast majority of studies across vertebrate taxa, phytohemagglutinin (PHA) is used as a universal stimulant. Given the complexity of immune response activation pathways of PHA, however, interpretation of test results can be ambiguous. Goal of this study was to improve methodology of the skin-swelling test to decrease this ambiguity. Here, we present an alternative protocol aimed at facilitating interpretation of skin-swelling data for mammals. Based on previous evidence suggesting that mammalian T cells are readily activated by Concanavalin A (ConA) in vitro, we compared cellular immune responses in vivo to PHA and ConA as an alternative pro-inflammatory stimulant in mice. We measured magnitude of tissue swelling and compared it with intensity of blood cell infiltration into tissue over a 72-hour interval. Our results corroborate that PHA and ConA show important differences in both dynamics and response amplitude in rodents. ConA induces stronger swelling with a distinct leukocyte activity pattern and higher pro-inflammatory cytokine (interleukin 6 [IL-6] and interferon gamma[IFN-γ]) expression than PHA during peak response (24-h post-treatment). Furthermore, unlike PHA, magnitude of swelling was positively associated with cellular activity (number of neutrophils infiltrating tissue) following ConA injection. We conclude that ConA is the more suitable stimulant for skin-swelling tests in mammals. This is because of the molecular binding specificity in the two lectins, that is, ConA specifically activates T cells while PHA also triggers erythroagglutination. We propose that ConA be used in all future ecological testing in mammals as it exhibits better performance and its application facilitates immunological interpretation of skin-swelling test results.
The aim of this study was to examine whether mesenchymal stem cells (MSCs) and/or corneal limbal epithelial stem cells (LSCs) influence restoration of an antioxidant protective mechanism in the corneal epithelium and renewal of corneal optical properties changed after alkali burns. The injured rabbit corneas (with 0.25 N NaOH) were untreated or treated with nanofiber scaffolds free of stem cells, with nanofiber scaffolds seeded with bone marrow MSCs (BM-MSCs), with adipose tissue MSCs (Ad-MSCs), or with LSCs. On day 15 following the injury, after BM-MSCs or LSCs nanofiber treatment (less after Ad-MSCs treatment) the expression of antioxidant enzymes was restored in the regenerated corneal epithelium and the expressions of matrix metalloproteinase 9 (MMP9), inducible nitric oxide synthase (iNOS), α-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), and vascular endothelial factor (VEGF) were low. The central corneal thickness (taken as an index of corneal hydration) increased after the injury and returned to levels before the injury. In injured untreated corneas the epithelium was absent and numerous cells revealed the expressions of iNOS, MMP9, α-SMA, TGF-β1, and VEGF. In conclusion, stem cell treatment accelerated regeneration of the corneal epithelium, restored the antioxidant protective mechanism, and renewed corneal optical properties.
- MeSH
- alkálie MeSH
- antioxidancia terapeutické užití MeSH
- buněčná diferenciace účinky léků MeSH
- chemické popálení enzymologie genetika patologie terapie MeSH
- imunohistochemie MeSH
- králíci MeSH
- limbus corneae cytologie MeSH
- matrixová metaloproteinasa 9 metabolismus MeSH
- mezenchymální kmenové buňky cytologie účinky léků MeSH
- ochranné látky farmakologie terapeutické užití MeSH
- pachymetrie rohovky MeSH
- regulace genové exprese účinky léků MeSH
- rohovkový epitel patologie MeSH
- superoxiddismutasa metabolismus MeSH
- synthasa oxidu dusnatého, typ II metabolismus MeSH
- transformující růstový faktor beta genetika metabolismus MeSH
- transplantace mezenchymálních kmenových buněk * MeSH
- tukové buňky cytologie účinky léků MeSH
- vaskulární endoteliální růstový faktor A metabolismus MeSH
- zákal rohovky komplikace terapie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- odvolaná publikace MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alkálie MeSH
- antioxidancia MeSH
- matrixová metaloproteinasa 9 MeSH
- ochranné látky MeSH
- superoxiddismutasa MeSH
- synthasa oxidu dusnatého, typ II MeSH
- transformující růstový faktor beta MeSH
- vaskulární endoteliální růstový faktor A MeSH
UNLABELLED: Stem cell-based therapy has become an attractive and promising approach for the treatment of severe injuries or thus-far incurable diseases. However, the use of stem cells is often limited by a shortage of available tissue-specific stem cells; therefore, other sources of stem cells are being investigated and tested. In this respect, mesenchymal stromal/stem cells (MSCs) have proven to be a promising stem cell type. In the present study, we prepared MSCs from bone marrow (BM-MSCs) or adipose tissue (Ad-MSCs) as well as limbal epithelial stem cells (LSCs), and their growth, differentiation, and secretory properties were compared. The cells were grown on nanofiber scaffolds and transferred onto the alkali-injured eye in a rabbit model, and their therapeutic potential was characterized. We found that BM-MSCs and tissue-specific LSCs had similar therapeutic effects. Clinical characterization of the healing process, as well as the evaluation of corneal thickness, re-epithelialization, neovascularization, and the suppression of a local inflammatory reaction, were comparable in the BM-MSC- and LSC-treated eyes, but results were significantly better than in injured, untreated eyes or in eyes treated with a nanofiber scaffold alone or with a nanofiber scaffold seeded with Ad-MSCs. Taken together, the results show that BM-MSCs' therapeutic effect on healing of injured corneal surface is comparable to that of tissue-specific LSCs. We suggest that BM-MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain. SIGNIFICANCE: Damage of ocular surface represents one of the most common causes of impaired vision or even blindness. Cell therapy, based on transplantation of stem cells, is an optimal treatment. However, if limbal stem cells (LSCs) are not available, other sources of stem cells are tested. Mesenchymal stem cells (MSCs) are a convenient type of cell for stem cell therapy. The therapeutic potential of LSCs and MSCs was compared in an experimental model of corneal injury, and healing was observed following chemical injury. MSCs and tissue-specific LSCs had similar therapeutic effects. The results suggest that bone marrow-derived MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain.
- Klíčová slova
- Alkali-injured ocular surface, Corneal regeneration, Limbal stem cells, Mesenchymal stem cells, Stem cell-based therapy,
- MeSH
- biologické markery metabolismus MeSH
- buněčná a tkáňová terapie metody MeSH
- buněčná diferenciace MeSH
- buňky kostní dřeně cytologie fyziologie MeSH
- chemické popálení patologie terapie MeSH
- epitelové buňky cytologie fyziologie transplantace MeSH
- exprese genu MeSH
- fyziologická neovaskularizace MeSH
- králíci MeSH
- limbus corneae krevní zásobení zranění MeSH
- mezenchymální kmenové buňky cytologie fyziologie MeSH
- primární buněčná kultura MeSH
- proliferace buněk MeSH
- reepitalizace fyziologie MeSH
- rohovkový epitel krevní zásobení zranění MeSH
- tkáňové podpůrné struktury MeSH
- transplantace mezenchymálních kmenových buněk * MeSH
- tuková tkáň cytologie fyziologie MeSH
- tukové buňky cytologie fyziologie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- biologické markery MeSH
The aim of this study was to investigate the effects of systemically administered bone-marrow-derived mesenchymal stromal cells (MSCs) on the early acute phase of inflammation in the alkali-burned eye. Mice with damaged eyes were either untreated or treated 24 h after the injury with an intravenous administration of fluorescent-dye-labeled MSCs that were unstimulated or pretreated with interleukin-1α (IL-1α), transforming growth factor-β (TGF-β), or interferon-γ (IFN-γ). Analysis of cell suspensions prepared from the eyes of treated mice on day 3 after the alkali burn revealed that MSCs specifically migrated to the damaged eye and that the number of labeled MSCs was more than 30-times higher in damaged eyes compared with control eyes. The study of the composition of the leukocyte populations within the damaged eyes showed that all types of tested MSCs slightly decreased the number of infiltrating lymphoid and myeloid cells, but only MSCs pretreated with IFN-γ significantly decreased the percentage of eye-infiltrating cells with a more profound effect on myeloid cells. Determining cytokine and NO production in the damaged eyes confirmed that the most effective immunomodulation was achieved with MSCs pretreated with IFN-γ, which significantly decreased the levels of the proinflammatory molecules IL-1α, IL-6, and NO. Taken together, the results show that systemically administered MSCs specifically migrate to the damaged eye and that IFN-γ-pretreated MSCs are superior in inhibiting the acute phase of inflammation, decreasing leukocyte infiltration, and attenuating the early inflammatory environment.
- MeSH
- alkálie toxicita MeSH
- alografty MeSH
- antivirové látky farmakologie MeSH
- chemické popálení patologie terapie MeSH
- interferon gama farmakologie MeSH
- interleukin-1alfa metabolismus MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nika kmenových buněk * MeSH
- popálení oka chemicky indukované metabolismus patologie terapie MeSH
- transformující růstový faktor beta metabolismus MeSH
- transplantace mezenchymálních kmenových buněk * MeSH
- zánět chemicky indukované metabolismus terapie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alkálie MeSH
- antivirové látky MeSH
- interferon gama MeSH
- interleukin-1alfa MeSH
- transformující růstový faktor beta MeSH
Regulatory T cells have been well described and the factors regulating their development and function have been identified. Recently, a growing body of evidence has documented the existence of interleukin-10 (IL-10) -producing B cells, which are called regulatory B10 cells. These cells attenuate autoimmune, inflammatory and transplantation reactions, and the main mechanism of their inhibitory action is the production of IL-10. We show that the production of IL-10 by lipopolysaccharide-stimulated B cells is significantly enhanced by IL-12 and interferon-γ and negatively regulated by IL-21 and transforming growth factor-β. In addition, exogenous IL-10 also inhibits B-cell proliferation and the expression of the IL-10 gene in lipopolysaccharide-stimulated B cells. The negative autoregulation of IL-10 production is supported by the observation that the inclusion of anti-IL-10 receptor monoclonal antibody enhances IL-10 production and the proliferation of activated B cells. The effects of cytokines on IL-10 production by B10 cells did not correlate with their effects on B-cell proliferation or on IL-10 production by T cells or macrophages. The cytokine-induced changes in IL-10 production occurred on the level of IL-10 gene expression, as confirmed by increased or decreased IL-10 mRNA expression in the presence of a particular cytokine. The regulatory cytokines modulate the number of IL-10-producing cells rather than augmenting or decreasing the secretion of IL-10 on a single-cell level. Altogether these data show that the production of IL-10 by B cells is under the strict regulatory control of cytokines and that individual cytokines differentially regulate the development and activity of regulatory T cells and IL-10-producing regulatory B cells.
- Klíčová slova
- B cells, autoregulation, cytokines, immunosuppression, interleukin-10 production,
- MeSH
- aktivace lymfocytů MeSH
- buněčná diferenciace * účinky léků MeSH
- cytokiny genetika metabolismus MeSH
- homeostáza MeSH
- interferon gama metabolismus MeSH
- interleukin-10 genetika metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- lipopolysacharidy farmakologie MeSH
- messenger RNA metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- proliferace buněk MeSH
- regulace genové exprese MeSH
- regulační B-lymfocyty účinky léků imunologie metabolismus MeSH
- regulační T-lymfocyty imunologie metabolismus MeSH
- rekombinantní proteiny metabolismus MeSH
- signální transdukce MeSH
- TNF-alfa metabolismus MeSH
- transformující růstový faktor beta1 metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cytokiny MeSH
- IFNG protein, human MeSH Prohlížeč
- IL10 protein, mouse MeSH Prohlížeč
- interferon gama MeSH
- interleukin-10 MeSH
- lipopolysacharidy MeSH
- messenger RNA MeSH
- rekombinantní proteiny MeSH
- TGFB1 protein, human MeSH Prohlížeč
- TNF-alfa MeSH
- transformující růstový faktor beta1 MeSH
Ocular surface defects represent one of the most common causes of impaired vision or even blindness. For treatment, keratoplasty represents the first choice. However, if corneal defects are more extensive and associated with a limbal stem cell (LSC) deficiency, corneal transplantation is not a sufficient therapeutic procedure and only viable approach to treatment is the transplantation of LSCs. When the LSC deficiency is a bilateral disorder, autologous LSCs are not available. The use of allogeneic LSCs requires strong immunosuppression, which leads to side-effects, and the treatment is not always effective. The alternative and perspective approach to the treatment of severe ocular surface injuries and LSC deficiency is offered by the transplantation of autologous mesenchymal stem cells (MSCs). These cells can be obtained from the bone marrow or adipose tissue of the particular patient, grow well in vitro and can be transferred, using an appropriate scaffold, onto the damaged ocular surface. Here they exert beneficial effects by possible direct differentiation into corneal epithelial cells, by immunomodulatory effects and by the production of numerous trophic and growth factors. Recent experiments utilizing the therapeutic properties of MSCs in animal models with a mechanically or chemically injured ocular surface have yielded promising results and demonstrated significant corneal regeneration, improved corneal transparency and a rapid healing process associated with the restoration of vision. The use of autologous MSCs thus represents a promising therapeutic approach and offers hope for patients with severe ocular surface injuries and LSC deficiency.
- MeSH
- autologní transplantace MeSH
- biologické modely MeSH
- buněčná diferenciace MeSH
- buňky kostní dřeně cytologie metabolismus MeSH
- CD antigeny metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- mezenchymální kmenové buňky cytologie metabolismus MeSH
- mezibuněčné signální peptidy a proteiny metabolismus MeSH
- nanovlákna * MeSH
- nemoci rohovky chirurgie MeSH
- pohyb buněk MeSH
- transplantace kmenových buněk metody MeSH
- transplantace mezenchymálních kmenových buněk metody MeSH
- tuková tkáň cytologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- CD antigeny MeSH
- mezibuněčné signální peptidy a proteiny MeSH