RESUMO
Reprogramming human somatic cells to pluripotency represents a valuable resource for research aiming at the development of in vitro models for human diseases and regenerative medicines to produce patient-specific induced pluripotent stem (iPS) cells. Seeking appropriate cell resources for higher efficiency and reducing the risk of viral transgene activation, especially oncogene activation, are of significance for iPS cell research. In this study, we tested whether human amnion-derived cells (hADCs) could be rapidly and efficiently reprogrammed into iPS cells by the defined factors: OCT4/SOX2/NANOG. hADCs from normal placenta were isolated and cultured. The 3rd passage cells were infected with the lentiviral vectors for the delivery of OCT4, SOX2, and NANOG. Afterwards, the generated iPSCs were identified by morphology, pluripotency markers, global gene expression profiles, and epigenetic status both in vitro and in vivo. The results showed that we were able to reprogram hADCs by the defined factors (OCT4/SOX2/NANOG). The efficiency was significantly high (about 0.1%), and the typical colonies appeared on the 9th day after infection. They were similar to human embryonic stem (ES) cells in morphology, proliferation, surface markers, gene expression, and the epigenetic status of pluripotent cell-specific genes. Furthermore, these cells were able to differentiate into various cell types of all three germ layers both in vitro and in vivo. These results demonstrate that hADCs were an ideal somatic cell resource for the rapid and efficient generation of iPS cells by OCT4/SOX2/NANOG.
Assuntos
Âmnio/citologia , Diferenciação Celular/fisiologia , Proteínas de Homeodomínio/fisiologia , Fator 3 de Transcrição de Octâmero/fisiologia , Células-Tronco Pluripotentes/citologia , Fatores de Transcrição SOXB1/fisiologia , Fosfatase Alcalina/metabolismo , Sequência de Bases , Western Blotting , Linhagem Celular , Primers do DNA , Imunofluorescência , Perfilação da Expressão Gênica , Proteínas de Homeodomínio/genética , Humanos , Proteína Homeobox Nanog , Fator 3 de Transcrição de Octâmero/genética , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase , Fatores de Transcrição SOXB1/genéticaRESUMO
OBJECTIVE: The purpose of this study was to gain a further understanding of the relationship between miR-152 and human leukocyte antigen (HLA)-G in human trophoblast cell line (JEG-3). STUDY DESIGN: The JEG-3 cells were transfected with pre-miR-152. The effect of the overexpressed miR-152 on HLA-G expression, trophoblast invasion, and natural killer (NK) cell-mediated cytolysis were assessed by reverse-transcription polymerase chain reaction (RT-PCR) and Western blot analysis, transwell invasion assay, and NK cell cytotoxicity assay, respectively. RESULTS: The miR-152 repressed HLA-G expression but exerted no effect on JEG-3 cell invasion, and overexpression of miR-152 led to increased NK cell-mediated cytolysis in JEG-3 cells. CONCLUSION: The data indicate that miR-152 may function as an immune system enhancer through up-regulating NK cell-mediated cytolysis of host cells.
Assuntos
Antígenos HLA/metabolismo , Antígenos de Histocompatibilidade Classe I/metabolismo , Células Matadoras Naturais/metabolismo , Trofoblastos/metabolismo , Análise de Variância , Western Blotting , Linhagem Celular , Células Cultivadas , Citotoxicidade Imunológica/genética , Citotoxicidade Imunológica/imunologia , Antígenos HLA/genética , Antígenos HLA/imunologia , Antígenos HLA-G , Antígenos de Histocompatibilidade Classe I/genética , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Células Matadoras Naturais/imunologia , MicroRNAs/genética , MicroRNAs/imunologia , MicroRNAs/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trofoblastos/citologia , Trofoblastos/imunologia , Regiões não Traduzidas/genética , Regiões não Traduzidas/imunologiaRESUMO
OBJECTIVE: To investigate the metabolic changes of calcium and phosphorus in dentin, dental pulp and periodontium in tail-suspended rats, and the functions of TGF-beta 1, c-fos, collagen-I and collagen IV in dentin, dental pulp and periodontium. METHOD: Relative percentage contents of Ca, P in dentin, dental pulp and periodontium were measured with scanning electron microscope and energy spectrum analytical system in 3 groups of rats. The expression of TGF-beta 1, c-fos, collagen-I and collagen IV were also observed. RESULT: In the suspension group, the relative percentage content of Ca declined significantly, while P increased slightly. There were no significant differences of Ca, P in alveolar bone. The expressions of TGF-beta 1, c-fos and collagen-I declined, but the expression of collagen-IV in pulp vessel increased. There were no significant changes of expressions of TGF-beta 1, c-fos, collagen-I and collagen-IV in the vicinity of PDL. After adopting artificial countermeasures, the above expressions restored partly. CONCLUSION: Weightlessness might cause abnormal mineralization in dentin, and 1.5 G artificial countermeasures could eliminate the above changes of mineral metabolism. The poor mineralization of dentin might be associated with the reduced secretion of TGF-beta 1, c-fos and collagen-I in tail-suspended rats.