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1.
Cell Tissue Bank ; 21(2): 233-248, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32052220

RESUMO

Injured nerves cannot regenerate on their own, and a lack of engraftable human nerves has been a major obstacle in cell-based therapies for regenerating damaged nerves. A monolayer culture approach to obtain adherent neural stem cells from human embryonic stem cells (hESC-NSCs) was established, and the greatest number of stemness characteristics were achieved by the eighth generation of hESC-NSCs (P8 hESC-NSCs). To overcome deficits in cell therapy, we used microvesicles secreted from P8 hESC-NSCs (hESC-NSC-MVs) instead of entire hESC-NSCs. To investigate the therapeutic efficacy of hESC-NSC-MVs in vitro, hESC-NSC-MVs were cocultured with dorsal root ganglia to determine the length of axons. In vivo, we transected the sciatic nerve in SD rats and created a 5-mm gap. A sciatic nerve defect was bridged using a silicone tube filled with hESC-NSC-MVs (45 µg) in the MVs group, P8 hESC-NSCs (1 × 106 single cells) in the cell group and PBS in the control group. The hESC-NSC-MVs group showed better morphological recovery and a significantly greater number of regenerated axons than the hESC-NSCs group 12 weeks after nerve injury. These results indicated that the hESC-NSC-MVs group had the greatest ability to repair and reconstruct nerve structure and function. As a result, hESC-NSC-MVs may have potential for applications in the field of nerve regenerative repair.


Assuntos
Micropartículas Derivadas de Células/metabolismo , Células-Tronco Embrionárias Humanas/metabolismo , Regeneração Nervosa/fisiologia , Células-Tronco Neurais/metabolismo , Nervo Isquiático/fisiologia , Animais , Animais Recém-Nascidos , Axônios/metabolismo , Biomarcadores/metabolismo , Diferenciação Celular , Linhagem Celular , Gânglios Espinais/metabolismo , Humanos , Músculos/fisiologia , Nanopartículas/química , Células-Tronco Neurais/citologia , Ratos Sprague-Dawley
2.
Stem Cells Int ; 2019: 6452684, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31772588

RESUMO

Myocardial reperfusion injury (MRI) induced by cardiomyocyte apoptosis plays an important role in the pathogenesis of a variety of cardiovascular diseases. New MRI treatments involving stem cells are currently being developed because these cells may exert their therapeutic effects primarily through paracrine mechanisms. Microvesicles (MVs) are small extracellular vesicles that have become the key mediators of intercellular communication. MVs derived from stem cells have been reported to play an important role in MRI. In this article, we attempted to explore the mechanisms by which MVs derived from human embryonic neural stem cells (hESC-NSC-derived MVs) rescue MRI. hESCs were differentiated into NSCs, and MVs were isolated from their supernatants by ultracentrifugation. H2O2 was used to induce apoptosis in HL-1 cardiomyocytes. Cell viability was detected by using the CCK-8 assay, apoptosis was detected by Annexin V-FITC/PI staining, and apoptosis-related proteins and signalling pathway-related proteins were detected by western blot analysis. Autophagic flux was measured using the tandem fluorescent mRFG-GFP-LC3 assay. Transmission electron microscopy and western blot analysis were adopted to evaluate autophagy levels. hESC-NSC-derived MVs increased the autophagy and inhibited the apoptosis of HL-1 cells exposed to H2O2 for 3 h in a dose-dependent manner. Additionally, hESC-NSC-derived MVs contained high levels of heat shock protein 70 (HSP-70), which can increase the level of HSP-70 in cells. Moreover, the same effect could be achieved by heat shock preconditioning of HL-1 cells overexpressing HSP-70. The benefits of NSC-MVs may be due to the involvement of AKT and mTOR signalling pathways. Importantly, hESC-NSC-derived MVs stimulated the activation of the AKTand mTOR signalling pathway in those cells by transporting HSP-70. Our results suggest that hESC-NSC-derived MVs inhibit the apoptosis of HL-1 cardiomyocytes by promoting autophagy and regulating AKT and mTOR via transporting HSP-70. However, this hypothesis requires in vivo confirmation.

3.
J Mater Sci Mater Med ; 30(7): 82, 2019 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-31273463

RESUMO

Axon regeneration and functional recovery after peripheral nerve injury remains a clinical challenge. Injury leads to axonal disintegration after which Schwann cells (SCs) and macrophages re-engage in the process of regeneration. At present, biomaterials are regarded as the most promising way to repair peripheral nerve damage. As a natural material, keratin has a wide range of sources and has good biocompatibility and biodegradability. Here, a keratin was extracted from human hair by reducing method and a keratin sponge with porous structure was obtained by further processing. The results suggested that keratin can promote cell adhesion, proliferation, migration as well as the secretion of neurotrophic factors by SCs and the regulation of the expression of macrophage inflammatory cytokines in vitro. We report for the first time that human hair keratin can promote the extension of axon in DRG neurons. The motor deficits caused by a sciatic nerve crush injury were alleviated by keratin sponge dressing in vivo. Thus, keratin has been identified as a valuable biomaterial that can enhance peripheral nerve regeneration.


Assuntos
Cabelo/química , Queratinas Específicas do Cabelo/farmacologia , Regeneração Nervosa/efeitos dos fármacos , Nervos Periféricos/efeitos dos fármacos , Nervo Isquiático/lesões , Animais , Axônios/efeitos dos fármacos , Materiais Biocompatíveis , Adesão Celular , Linhagem Celular , Movimento Celular , Proliferação de Células , Citocinas/metabolismo , Humanos , Inflamação , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Neurônios/metabolismo , Células RAW 264.7 , Ratos , Ratos Sprague-Dawley , Células de Schwann/efeitos dos fármacos , Cicatrização
4.
Oncol Rep ; 38(2): 1013-1020, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28627682

RESUMO

Human embryonic stem cell derived-mesenchymal stem cells (hESC­MSCs) are able to inhibit proliferation of leukemia cells. Microvesicles released from human embryonic stem cell derived-mesenchymal stem cells (hESC­MSC­MVs) might play an important part in antitumor activity. Microvesicles were isolated by ultracentrifugation and identified under a scanning electron microscopy and transmission electron microscope separately. After 48-h cocultured with hESC­MSCs and hESC­MSC­MVs, the number of K562 and HL60 was counted and tumor cell viability was measured by CCK8 assay. The expression of proteins Bcl-2 and Bax were estimated by western blotting. Transmission electron microscope and western blot analysis were adopted to evaluate the autophagy level. Results showed that both hESC­MSCs and hESC­MSC­MVs inhibited proliferation of leukemia cells in a concentration-dependent manner. hESC­MSC­MVs reduced the ratio of Bcl/Bax, enhanced the protein level of Beclin-1 and LC3-II conversion, thus upregulating autophagy and apoptosis. In conclusion, microvesicles released from human embryonic stem cell derived-mesenchymal stem cells inhibited tumor growth and stimulated autophagy and excessive autophagy might induce apoptosis.


Assuntos
Apoptose , Proliferação de Células , Micropartículas Derivadas de Células/patologia , Células-Tronco Embrionárias Humanas/citologia , Leucemia/patologia , Células-Tronco Mesenquimais/citologia , Autofagia , Micropartículas Derivadas de Células/metabolismo , Células Cultivadas , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Leucemia/metabolismo , Células-Tronco Mesenquimais/metabolismo
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