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- ItemSomente MetadadadosEfeito da célula-tronco mesenquimal e do seu meio condicionado na lesão renal aguda induzida pela sepse e pelo aciclovir(Universidade Federal de São Paulo (UNIFESP), 2015-04-30) Christo, Joelma Santina [UNIFESP]; Schor, Nestor Schor [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Sepse é uma condição médica que afeta 18 milhões de pessoas por ano no mundo, é caracterizada por um estado inflamatório generalizado causado por uma infecção. A ativação generalizada de vias de coagulação e inflamação evolui para disfunção de múltiplos órgãos, o colapso do sistema circulatório (choque séptico) e morte. Apesar de décadas de pesquisas e numerosas experimentações clínicas, pouco progresso tem sido observado no desenvolvimento de novos tratamentos e as taxas de mortalidade são praticamente as mesmas nos últimos 20 a 30 anos. Recentemente as células-tronco têm emergido como uma terapia promissora para uma variedade de patologias, incluindo doenças cardiovasculares, neurodegenerativas, doença vascular periférica, doença renal, e várias outras. Portanto nesse trabalho avaliamos os efeitos renoprotetores das CTMS e do MC utilizando modelo de sepse induzida por Escherichia Coli em ratos. Ambos os tratamentos CTMs e MC exerceram uma ação renopretora evidenciado pela melhora dos parâmetros renais (Cr, U, Na+ e K+), seguido de alterações tubulares mais sutis nos animais que receberam CTMs ou MC quando comparados aos animais não tratados. Avaliamos também o efeito das CTMs ou do MC sob a resposta inflamatória desencadeada pela sepse e observamos uma diminuição da resposta inflamatória nesses animais, caracterizado por uma diminuição das citocinas pró-inflamatórias (IL-1?, IL-1?, IL-6, TNF-? e IFN-?) seguido de um aumento de IL-10 nesses animais, portanto esses efeitos não foram observados nos animais que não receberam CTMs ou MC. As alterações tubulares e glomerulares nesses animais também foram mais brandas, porém não significantes em relação aos animais não tratados com CTMs ou MC. Surpreendentemente não encontramos as células tronco nos rins dos animais tratados com CTMs sugerindo assim que os efeitos benéficos das CTMs ou do MC até então observados em nosso trabalho, são via ação parácrina das CTMs.
- ItemAcesso aberto (Open Access)Estabelecimento e caracterização de cultura primária de tecido vaginal. Viabilidade do uso de terapia celular no tratamento de pacientes com síndrome de Mayer-Rokitansky-Küster-Hauser(Universidade Federal de São Paulo (UNIFESP), 2015-12-31) Paula, Tatiane Aparecida de [UNIFESP]; Girão, Manoel Joao Batista Castello [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)The uterine and vaginal agenesis, also known as Syndrome Mayer-Rokitansky-Küster-Hauser (SMRKH) is a congenital malformation that, although rare, carries important clinical and psychological repercussions, due to the impairment of sexual activity and reproductive life. The treatment for this disease is divided into bloodless and surgical methods and consists in the construction of a neovagina, allowing the patient to have a normal sexual life. However, there is no consensus on best approach of treatment because of the advantages and disadvantages presented in both cases. Recently experimental studies described a successful technique of rabbit vaginal tissue engineering using samples of vagina and muscle autologous cells. In 2007 it was reported the first neovaginoplasty case with the MacIndoe modified technique were it was used the patient's own cells to produce an autologous vaginal tissue in the laboratory. Objectives: Our aim is to use the patient's own cells to produce an autologous vaginal tissue in the laboratory to be used in the near future in neovaginoplasty. Methods: We performed 05 full-thickness biopsies from vaginal vestibule of ~ 1cm2. After several washes in PBS with antibiotics, the tissue was minced into small pieces which were then put in culture plates with DMEM/F-12 supplemented with 10% of fetal bovine serum to allow cells to migrate from the explants. After cells reached 70% of confluence, the explants were removed so cells could grown to remaining spaces, at this moment the plates were harvested and cells were frozen in this first passage at density of 106 cells/vial, if we could not obtain this cell density, cells were further expanded and frozen at second passage. Using flow citometry, 03 samples were characterized using the following antibodies: CD90, CD73, CD105 and STRO-I for mesenchymal stem cells as well as pan-citokeratyn, CD133, CD9, CD227 and CD1d for epithelial cells, being the last two specifically used for vaginal epithelial cells. Finally we used CD34 for hematopoietic progenitor cells and vimentin for fibroblasts. The data was acquired using FACS Canto II ?BD Bioscience and analyzed with FlowJo 7.6.5 software. Results: Using explant technique we were able to obtain a mixed cell population of viable cells. These cells were characterized by 67% of mesenchymal stem cells (CD90+ CD73+ CD105+; SD=10,18,); we find high positivity of STRO-I cells (75,2%, SD=8,77) and vimetin (80,3%, SD=4,59). For most epithelial markers used we also find high values: 71,9% of CD133, 73,7% of CD9 and 80,3% of CD227 (SD= 6,7, 5,74 and 8,77 respectively). We found a weak positivity of citokeratyn and PDGFR-RB cells among the samples (13,6%, 0,52%, and 1,96%; 0,93%, 0,38%, 0,20% respectively) and a weak positivity of CD34 cells (14,7%, SD= 4,98). Conclusions: Using explant technique we were able to establish primary cell cultures from vaginal biopsies. Those cultures are characterized by having a mixed population of cells, namely, stem cells, epithelial cells and fibroblasts, showing that this approach could be used to obtain autologous vaginal tissue.
- ItemAcesso aberto (Open Access)Expressão do gene da leptina em cultura de células de camundongos ob/ob como modelo para síndrome do ovário policístico(Universidade Federal de São Paulo (UNIFESP), 2014-10-29) Parreira, Rafael Martins [UNIFESP]; Patriarca, Marisa Teresinha [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)The polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women of reproductive age and affects 15 to 20% of patients with infertility. PCOS shows reproductive implications, endocrine, dermatological, gynecological, cardiac and psychological with symptoms ranging from infertility due to ovulatory dysfunction, menstrual disorders and / or androgenismo signals. Furthermore, obesity affects the majority of women with PCOS and therefore many studies have been done to relate obesity and infertility in women with PCOS. The weight loss can improve the levels of androgens and glucose, as well as ovulation and pregnancy rates. It is known that the energy balance in mice is regulated by the ob gene which expresses the leptin. This protein acts in adipose tissue by regulating the size of the body fat deposit, and play a role in reproduction, including a direct effect on the ovary. With advances in molecular biology techniques and the emergence of gene therapy as a new form of treatment, this study aims to verify the potential of gene therapy using the leptin gene in ob mice cells / ob as a model for PCOS. mouse cells ob / ob began to express the leptin gene after transfection. The expression levels of leptin in the transfected cells, both ob / ob and the control were significantly higher and above the baseline level. Given these results, there is the need for future in vivo studies for further treatment associated gene therapy and cell therapy in patients with infertility related to PCOS
- ItemAcesso aberto (Open Access)Sangue de cordão umbilical para uso autólogo ou grupo de pacientes especiais(Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular, 2009-05-01) Cruz, Luis Eduardo; Jorge, Maria Claudia; Machado, Janaína J.; Hossne Junior, Nelson Americo [UNIFESP]; Invitti, Adriana L.; Balduino, Alex; Vianna, Verônica; Ellovitch, Saada R. S.; Urago, Kátia P. T.; Meneses-Costa, Ana Paula; Ribeiro, Márcia G.; Nicola, Maria Helena A.; Sanberg, Paul R.; Cryopraxis Criobiologia Ltda; Universidade Federal de São Paulo (UNIFESP); Instituto de Traumato-ortopedia; Santa Casa de São Paulo; Hospital Samaritano de São Paulo; UFRJ HUCFF; Hospital Central do Exército; UFRJ IPPMG; UFRJ; College of Medicine Department of Neurosurgery Center of Excellence for Aging & Brain RepairUmbilical Cord Blood is a rich source of hematopoietic stem cells widely used as a substitute of bone marrow (BM) in transplants. Cells from umbilical cord blood present advantages over BM cells, mainly as they are younger and a have higher proliferative rate. Besides hematopoietic stem cells, umbilical cord blood contains endothelial and mesenchymal progenitor cells, suggesting their possible application in cell therapy protocols for different tissues. In this paper, we discuss the importance of autologous umbilical cord blood storage and the research on stem cell transplantation for degenerative diseases.