Navegando por Palavras-chave "Cell culture"
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- ItemAcesso aberto (Open Access)Caracterização molecular e funcional das células do endométrio de mulheres com endometriose pélvica(Universidade Federal de São Paulo (UNIFESP), 2016-04-27) Nogueira, Adriana Luckow Invitti [UNIFESP]; Girão, Manoel João Batista Castello [UNIFESP]; http://lattes.cnpq.br/0973903299568770; http://lattes.cnpq.br/2547404412649527; Universidade Federal de São Paulo (UNIFESP)The endometrium contains stem cells that possibly contribute to the regeneration of the functional layer during the mensal cycling. Different kinds of stem cells were isolated from endometrium and endometriotic implants. The monoclonal origin of endometriotic implants indicates that stem cells may have a role in the endometriosis pathogenesis. We isolated a mixed cells pool containing at least one stem cells population from healthy and endometriosis endometrium. These cells presented the markers CD90, CD73, CD105, CD44 and CD146, being able to differentiate into adipogenic, osteogenic and chondrogenic lineages and not responding to progesterone stimulus. The endometriosis stem cells expressed the CD34 naïve stem cells marker, presented higher proliferative rates than healthy ones and were able to form long lasting cells aggregates (spheroids) in tridimensional cultures. The endometrium with endometriosis had a unique stem cell population, reinforcing the hypothesis that stem cells play a key role in the endometriosis pathogenesis. 2. INTRODUCTION Stem cells are undifferentiated cells defined by their functional ability to self-renew and to differentiate into multiple cell lineages, as well as by plasticity and clonogenic potential. They were found abundantly in embryonic and fetal tissues and are also present in small amounts in the majority of the adult tissues; differing between each other only in the clonogenic and differentiation potential. Adult stem cells occur in niches that balance self-renewal with lineage selection and progression during tissue homeostasis
- ItemAcesso aberto (Open Access)Caracterização morfológica e molecular das endometriosferas formadas por células derivadas do endométrio de mulheres com endometriose profunda(Universidade Federal de São Paulo, 2024-02-05) Nogueira, Pollyana Telles [UNIFESP]; Nogueira, Adriana Luckow Invitti [UNIFESP]; Schor, Eduardo [UNIFESP]; http://lattes.cnpq.br/8854353153245040; http://lattes.cnpq.br/2547404412649527; http://lattes.cnpq.br/8492689839290665Endometriose é uma doença ginecológica crônica e benigna que se caracteriza pela presença de tecido semelhante ao endométrio fora da cavidade uterina. Estima-se que 10-15% das mulheres em idade reprodutiva apresentam a doença e a maioria experimenta grande diminuição na qualidade de vida. Dentre os sintomas, destacam-se a dismenorreia, a dispareunia e a infertilidade. Inúmeras teorias tentam explicar a fisiopatologia da doença, porém nenhuma é capaz de explicar todas as suas apresentações. O início da doença está diretamente relacionado à capacidade de sobrevivência e proliferação das células regurgitadas pelas tubas uterinas na cavidade peritoneal. Neste trabalho, avaliamos o papel das células endometriais estromais (eSCs) e epiteliais (eECs) na formação de esferoides em um modelo in vitro sem matriz de suporte mimetizando as células endometriais regurgitadas no fluido peritoneal. eSCs e eECs primárias foram isoladas do endométrio eutópico de mulheres com endometriose rASRM estágio IV e co-cultivadas em diferentes proporções desses tipos celulares. A morfologia dos esferoides e a participação de cada tipo celular foram avaliadas por microscopia. As eSCs são o principal componente dos esferoides em todas as proporções testadas e estão localizados na parte interna das esferas. O aumento das proporções de eSCs é diretamente proporcional ao aumento do tamanho dos esferoides. As células foram polarizadas, ficando as eSCs na parte interna do esferoide e as eECs revestindo a superfície. O marcador de células-tronco mesenquimais CD146 foi identificado até mesmo nas células da superfície do esferoide. Em conclusão, tanto as eECs como as eSCs contribuíram ativamente para a formação de esferoides endometriais. As interações entre eSCs e eECs são necessárias para a formação de esferoides estáveis e reforçam o possível papel do das interações entre eECs e eSCs para a sobrevivência de células endometriais
- ItemAcesso aberto (Open Access)Determinação de padrões de crescimento de células em cultura(Sociedade Brasileira de Patologia ClínicaSociedade Brasileira de PatologiaSociedade Brasileira de Citopatologia, 2003-01-01) Vilela, Marcelo José; Martins, Marcelo Lobato; Mendes, Rosemairy Luciane [UNIFESP]; Santos, Anésia Aparecida Dos; Universidade Federal de Viçosa Departamento de Biologia Animal; Universidade Federal de Viçosa Departamento de Física; Universidade Federal de São Paulo (UNIFESP); UFVThe growth patterns of established normal and cancer cell lines, cultured in monolayer and collagen gel, have been characterized using the cluster size distribution of cellular aggregates. HN-5 (cancer) cells exhibit, either in gel or in monolayer, power-law distributions at any time in culture, whereas for MDCK (normal) and HEp-2 (cancer) cells there is a transition from an exponential behavior to a power-law distribution after a transient time in culture. These results suggest that the transitions in growth regimes observed in MDCK and HEp-2 cell lines might be associated to changes in the control of replication or in the expression patterns of cell adhesion molecules of cell-cell and cell-matrix type related to intracellular signalling. These transitions are irreversible and seems to be an adaptative response to the growth constraints imposed by high cell population density or long permanence in culture.
- ItemAcesso aberto (Open Access)Endotélio venoso: estabelecimento e caracterização de culturas primárias e a produção de fatores derivados do endotélio de veia cava e veia porta de ratos(Universidade Federal de São Paulo, 2017-03-17) Trindade, Marcio Renato [UNIFESP]; Fernandes, Liliam [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)The endothelial cell plays an essential role in vascular control by the release of Endothelium- Derived Relaxing Factors (EDRFs) [nitric oxide (NO), prostacyclin (PGI2) and endotheliumderived hyperpolarizing factor (EDHF)], and Endothelium-Derived Constrictor Factors (EDCFs) [prostaglandin H2 (PGH2) and Thromboxane A2 (TXA2), reactive oxygen species, Angiotensin II (Ang II) and Endothelin-1 (ET-1)]. Most of the studies related to EDRFs and EDCFs refers to arterial territories and little is known about distinct venous territories, such as vena cava (VC) and portal vein (PV). The present study investigated the physiology of the venous endothelial cell using primary cultures of VC and PV of rats. Venous segments were sectioned longitudinally, plated with the endothelial side facing down, covered with culture medium and kept in a 5% CO2 incubator at 37°C for 5 days. After removal of the explants, the cells were subcultured and cultures were studied between 4th and 5th passages. The characterization of the cultures was performed by immunocytochemistry for the specific markers von Willebrand Factor (vWF) and endothelial NO synthase (eNOS), and positive staining was also observed for ET-1 receptor (ETB) and Ang II receptors (AT1 and AT2). Basal NO production was observed in endothelial cultures pre-incubated with the DAF-2DA fluorescent probe and observed in confocal microscopy, and complemented by eNOS expression analysis determined by western blot (n = 4). Prostanoids production was quantified in the culture supernatants by enzyme-immunoassay technique (n = 5 to 6). Endothelial cultures of VC and PV showed similar values in the baseline rates of PGH2 (0.75 ± 0.04 ng/mL vs 0.76 ± 0.02 ng / mL, respectively); these levels were increased after stimulation with Ang II [1μM] (1.00 ± 0.07 * ng/mL for VC vs 1.13 ± 0.07 * ng/mL for PV). Baseline levels of PGI2 were different between cultures, with values of 29.12 ± 1.24 ng/mL for VC vs 35.10 ± 1.27 * ng/mL for PV. Ang II stimulation [1μM] increased PGI2 production in VC cells (36.50 ± 1.89 * ng/mL) but not in PV (37.70 ± 1.38 ng/mL) (* P <.05); however, COX expression (1 and 2) was similar between cultures (determined by western blot, n = 7). Stimulation with ET-1 [1μM] did not modify the release of prostanoids in both cultures. In summary, the venous endothelium can be isolated and studied through primary cultures; these cells, when studied in vitro, produce detectable amounts of NO and prostaglandins. The participation of PGI2 in the venodilation can be pronounced in PV when compared to VC. The consistent increase in PHG2 production by Ang II in VC and PV endothelium indicates the important endothelial modulation in the venoconstriction. The present data reveal new aspects of the vascular physiology, and may contribute to a better understanding of the venous responses to the blocking agents of the Renin Angiotensin System.
- ItemAcesso aberto (Open Access)Experimental model for fibroblast culture(Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia, 2004-12-01) Keira, Sidney Mamoru [UNIFESP]; Ferreira, Lydia Masako [UNIFESP]; Gragnani, Alfredo [UNIFESP]; Duarte, Ivone da Silva [UNIFESP]; Santos-Vial, Isabel Anunciação Neves dos [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)The use of cell culture methods in Plastic Surgery opened a new horizon in the research of cellular mechanisms of proliferation and biosynthesis functions. Several types of cells have been investigated in the cutaneous compartment. Keratinocytes and fibroblasts have been studied aiming the possibility of developing biomaterial for skin substitution. The present study describes the standardization for the development of fibroblast primary culture, its utilization in experiments and its storage.
- ItemAcesso aberto (Open Access)Isolation, cultivation and characterization of CD133+ stem cells from human glioblastoma(Instituto Israelita de Ensino e Pesquisa Albert Einstein, 2012-06-01) Pavon, Lorena Favaro; Marti, Luciana Cavalheiro; Sibov, Tatiana Tais; Miyaki, Liza Aya Mabuchi; Malheiros, Suzana Maria Fleury [UNIFESP]; Mamani, Javier Bustamante; Brandt, Reynaldo Andre; Ribas, Guilherme Carvalhal; Pagura, Jorge Roberto; Joaquim, Marcos Augusto Stavale; Feres Junior, Hallin; Gamarra, Lionel Fernel; Hospital Israelita Albert Einstein Instituto do Cérebro; Hospital Israelita Albert Einstein Centro de Pesquisa Experimental; Hospital Israelita Albert Einstein Faculdade de Enfermagem; Universidade Federal de São Paulo (UNIFESP); Hospital Israelita Albert Einstein Center for Neuro-oncology; Hospital Israelita Albert EinsteinOBJECTIVE: To establish the method of isolation and culture of human glioblastoma neurospheres, and the purification of their stem cells, followed by the process of obtaining tumor subspheres, immunophenotypically characterizing this clonogenic set. METHODS: Through the processing of glioblastoma samples (n=3), the following strategy of action was adopted: (i) establish primary culture of glioblastoma; (ii) isolation and culture of tumor neurospheres; (iii) purify cells that initiate tumors (CD133+) by magnetic separation system (MACS); (iv) obtain tumor subspheres; (v) study the expression of the markers nestin, CD133, and GFAP. RESULTS: The study successfully described the process of isolation and culture of glioblastoma subspheres, which consist of a number of clonogenic cells immunophenotypically characterized as neural, which are able to initiate tumor formation. CONCLUSION: These findings may contribute to a better understanding of the process of gliomagenesis.
- ItemSomente MetadadadosThe metastatic behavior of osteosarcoma by gene expression and cytogenetic analyses(Elsevier B.V., 2013-10-01) Salinas-Souza, Carolina [UNIFESP]; De Oliveira, Renato [UNIFESP]; De Seixas Alves, Maria Teresa [UNIFESP]; Garcia Filho, Reynaldo Jesus [UNIFESP]; Petrilli, Antonio Sergio [UNIFESP]; Toledo, Silvia Regina Caminada de [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Osteosarcoma is a malignant bone tumor with high metastatic potential. Metastasis at diagnosis is the most significant prognostic factor in predicting the clinical outcome of osteosarcoma. We compared the gene expression of metastases that were present at the time of initial diagnosis to those developed later in the course of the disease. We used quantitative real-time polymerase chain reaction to evaluate the gene expression of MDM2, CXCR4, RANKL, RB1, and OSTERIX in 98 samples of osteosarcoma taken from 47 patients (74 metastases and 24 primary tumors) and 30 nonmalignant lung tissues surrounding osteosarcoma metastases. in addition, we investigated the copy number changes of RB1 and MDM2 genes in 12 primary cultures of pulmonary metastases of osteosarcoma, using interphase fluorescence in situ hybridization. Metastases from metastatic patients at diagnosis were characterized by low expression of RB1 and RANKL (P = .0009 and P = .0109, respectively) and overexpression of CXCR4 and MDM2 (P = .0389 and P = .0325, respectively). the loss of RANKL and gain of CXCR4 could also be detected in the primary tumors of metastatic patients at diagnosis (P = .0121 and P = .0264, respectively). Thus, some early genetic events such as the loss of RANKL and the gain of CXCR4 expressions probably facilitate the metastatic progression concomitant with the primary tumor establishment, supporting the role of the CXCR4 receptor in directing osteosarcoma metastases to the lung. On the other hand, late events such as the loss of RB1 and gain of MDM2, crucial regulators of cell cycle, appear to be related to the final mechanisms contributing to the metastatic establishment of osteosarcoma. (c) 2013 Elsevier Inc. All rights reserved.
- ItemSomente MetadadadosRetinal and Ocular Toxicity in Ocular Application of Drugs and Chemicals - Part I: Animal Models and Toxicity Assays(Karger, 2010-01-01) Penha, Fernando Marcondes [UNIFESP]; Rodrigues, Eduardo B. [UNIFESP]; Maia, Mauricio [UNIFESP]; Dib, Eduardo [UNIFESP]; Costa, Elaine Fiod [UNIFESP]; Furlani, Bruno A. [UNIFESP]; Moraes Filho, Milton Nunes [UNIFESP]; Dreyfuss, Juliana L. [UNIFESP]; Bottos, Juliana [UNIFESP]; Farah, Michel E. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Aims: Experimental retinal research has gained great importance due to the ophthalmic pharmacotherapy era. An increasing number of drugs are constantly released into the market for the treatment of retinal diseases. in this review, animal species, animal models and toxicity assays in retinal research are discussed. Methods: An extensive search of the literature was performed to review various aspects of the methods of investigation of drug toxicity. the different types of animal species, as well as single animal models available for the evaluation of safety and efficacy of retinal pharmacotherapy, were identified. in addition, a large variety of reported laboratory techniques were critically examined. Results: in vitro studies are the first-line experiments for the development of a new drug for retinal diseases, using retinal pigment epithelial cells and other cell lines. the next step involves in vivo animal studies where nonhuman primates are considered the gold standard. However, cost and legal issues make their use difficult. Mice and rats provide genetically controlled models for investigations. Pigs, dogs and cats represent good large-size animal models, while rabbits are one of the most used species for retinal toxicity evaluations. Various laboratory methods were identified, including light microscopy, electron microscopy, electroretinography and new emerging methods, such as optical coherence tomography and scanning laser ophthalmoscopy for experimental purposes. Conclusions: A great number of animal species and models are available that simulate retinal diseases and provide experimental data for further human use. Work with animal models should include properly designed toxicity assays to obtain reliable results for safety and efficacy. Copyright (C) 2010 S. Karger AG, Basel
- ItemSomente MetadadadosVenous endothelium reactivity to Angiotensin II: A study in primary endothelial cultures of rat vena cava and portal vein(Elsevier Inc, 2018) Trindade, Marcio Renato [UNIFESP]; Reis Assuncao, Henrique Charlanti [UNIFESP]; Torres, Tathiany Corteze [UNIFESP]; Bertolino, Jessica Silva [UNIFESP]; Fernandes, Liliam [UNIFESP]The role of the vascular endothelium in modulating the arterial system has been widely investigated, but poorly explored at the venous site. In the present work, primary cultures of venous endothelium from rat Vena Cava (VC) and Portal Vein (PV) were established, characterized and analyzed according to their growth pattern and ability to produce nitric oxide (NO) and prostanoids (PGF(2) (alpha) and PGI(2)), at basal state and after stimulation with Angiotensin II (Ang II, 1 mu mol/L). Basal NO was detected in all examined cells in culture. Pre-incubation with Ang II increased NO production in cells from VC (but not in PV cultures), through activation of both AT(1). and AT(2) receptors. Both cultures exhibited detectable levels of PGF(2) (alpha) at resting conditions, which were similarly enhanced by Ang II. Basal PGI(2) levels were higher in PV, but increased after Ang II treatment in VC, with no further effect on PV cells. We conclude that endothelial cells from VC and PV exhibit important properties and react to Ang II, probably influencing the whole circulatory system. This experimental cell model gives support to further studies concerning intracellular pathways of the venous endothelium, analyzed in separate from the vascular smooth muscle wall.