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Comparison of direct immunofluorescence, conventional cell culture and polymerase chain reaction techniques for detecting respiratory syncytial virus in nasopharyngeal aspirates from infants; Comparação entre imunofluorescência direta, cultura convencional de células e reação em cadeia de polimerase para detectar vírus respiratório sincicial em lavados de nasofaringe de crianças

REIS, Alexanda Dias; FINK, Maria Cristina Domingues; MACHADO, Clarisse Martins; PAZ JR., José de Paula; OLIVEIRA, Renato Reis; TATENO, Adriana Fumie; MACHADO, Adriana Freire; CARDOSO, Maria Regina; PANNUTI, Claudio Sérgio; CHIADO; FAPESP Research Groups
Fonte: Instituto de Medicina Tropical Publicador: Instituto de Medicina Tropical
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
55.74%
A total of 316 samples of nasopharyngeal aspirate from infants up to two years of age with acute respiratory-tract illnesses were processed for detection of respiratory syncytial virus (RSV) using three different techniques: viral isolation, direct immunofluorescence, and PCR. Of the samples, 36 (11.4%) were positive for RSV, considering the three techniques. PCR was the most sensitive technique, providing positive findings in 35/316 (11.1%) of the samples, followed by direct immunofluorescence (25/316, 7.9%) and viral isolation (20/315, 6.3%) (p < 0.001). A sample was positive by immunofluorescence and negative by PCR, and 11 (31.4%) were positive only by RT-PCR. We conclude that RT-PCR is more sensitive than IF and viral isolation to detect RSV in nasopharyngeal aspirate specimens in newborn and infants.; Um total de 316 amostras de lavado de nasofaringe obtidas de crianças em acompanhamento ambulatorial com até dois anos de idade durante episódio de doença aguda do trato respiratório foram processadas para detecção do vírus sincicial respiratório (VSR) utilizando três diferentes técnicas: isolamento viral, imunofluorescência direta e reação em cadeia por polimerase (RT-PCR). Destas amostras, 36 (11,4%) foram positivas para o VSR. A RT-PCR foi a técnica mais sensível...

Comparison of two cellular harvesting methods for primary human oral culture of keratinocytes

KLINGBEIL, Ma Fatima Guarizo; HERSON, Marisa Roma; CRISTO, Elier Broche; PINTO JR., Decio dos Santos; YOSHITO, Daniele; MATHOR, Monica Beatriz
Fonte: SPRINGER Publicador: SPRINGER
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
55.83%
The possibility of obtaining transplantable oral epithelia opens new perspectives for oral treatments. Most of them are surgical, resulting in mucosal failures. As reconstructive material this in vitro epithelia would be also useful for other parts of the human body. Many researchers still use controversial methods; therefore it was evaluated and compared the efficiency of the enzymatic and direct explant methods to obtain oral keratinocytes. To this project oral epithelia fragments were used. This work compared: time needed for cell obtainment, best cell amount, life-span and epithelia forming cell capacity. The results showed the possibility to obtain keratinocytes from a small oral fragment and we could verify the advantages and peculiar restrictions. We concluded that under our conditions the enzymatic method showed the best results: in the cells obtaining time needed, cell amount and life-span. Both methods showed the same capacity to form in vitro epithelia.

An in vitro study showing the three-dimensional microenvironment influence over the behavior of head and neck squamous cell carcinoma

Giudice, Fernanda Salgueiredo; Abrahão, Aline Corrêa; Sperandio, Felipe Fornias; Vechio, Aluana Maria da Costa Dal; Pinto-Junior, Decio dos Santos
Fonte: MEDICINA ORAL S L; VALENCIA Publicador: MEDICINA ORAL S L; VALENCIA
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
55.76%
Objectives: The Head and Neck Squamous Cell Carcinoma (HNSCC) ranks sixth worldwide. The mechanisms of growth, invasion and metastasis of this pathology are extensively studied and generally related to specific variations in signaling pathways like the PI3K-Akt; however most of these competent studies have been performed bidimensionally, which may hide important questions. This study sought to analyze the influence of the microenvironment upon the behavior of HNSCC. Study Design: The status of pAkt, NF-kappa B and Cyclin D1 proteins was accessed through immunofluorescence and western blot methods in HNSCC cell lines originating from tongue, pharynx and metastatic lymph node when submitted to a three-dimensional culture model utilizing a matrix system. A bidimensional culture model (monolayer) was used as control. Results: The HNSCC cell lines cultured three-dimensionally exhibited a growth pattern characterized by small isolated islands, different from the control group. When the three-dimensional model was applied, two of the studied cell lines showed the same expression pattern as the bidimensional model regarding nuclear or cytoplasmatic localization, as well as reduction of all protein levels; however, the cell line originated from tongue...

"Comparação de dois métodos de obtenção celular para cultura primária de queratinócitos bucais humanos" ; The comparison of two methods to obtain human oral keratinocytes in primary culture

Klingbeil, Maria Fátima Guarizo
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 21/11/2006 Português
Relevância na Pesquisa
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Freqüentemente as condutas terapêuticas utilizadas no tratamento de patologias bucais são cirúrgicas, resultando em falhas de continuidade da mucosa bucal. A possibilidade de obtenção de epitélios transplantáveis, a partir do cultivo in vitro de células da mucosa bucal, abre novas perspectivas de utilização, não se restringindo somente ao seu local de origem, ou seja, a boca, mas também como material de reconstrução para outras regiões, tais como: uretra, córnea, superfície ocular e epitélio córneo-limbal. Os métodos utilizados para a obtenção dessas células ainda são controversos na literatura. Neste sentido, avaliamos e comparamos a eficiência de dois métodos, enzimático e explante, para a obtenção de queratinócitos de mucosa bucal humana. Os fragmentos utilizados para a obtenção dessas células foram obtidos durante procedimentos cirúrgicos de pacientes voluntários saudáveis. Os queratinócitos foram cultivados sobre uma camada de sustentação, feeder-layer, confeccionada com fibroblastos murinos irradiados (3T3 - Swiss albino). Neste estudo foram comparados: o tempo para a obtenção dos queratinócitos, o rendimento obtido entre os dois métodos, a duração da vida útil em cultura, a capacidade que estas células tiveram em formar um epitélio in vitro e a morfologia dos mesmos. Os resultados obtidos...

Padronização de técnicas de isolamento de células de Langerhans imaturas e desenvolvimento de um modelo tridimensional de pele humana para testes de sensibilidade in vitro; Standardization of techniques for isolation of immature Langerhans cells and development of a three-dimensional human skin model for in vitro sensibility tests

Luco, Dayane Piffer
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 18/09/2014 Português
Relevância na Pesquisa
55.75%
A pele é o maior órgão do corpo humano e constitui a principal defesa do organismo contra agentes físicos e químicos, sendo também fundamental para evitar a perda de água por dessecação. Formada por três camadas distintas, mas complementares, sendo as duas principais denominadas derme e epiderme, contendo diferentes tipos celulares, como fibroblastos, queratinócitos, melanócitos, células de Merkel e células de Langerhans, sendo que estas últimas desempenham um papel fundamental na hipersensibilidade de contato. Devido à importância da manutenção da pele saudável para a vida humana, existe uma crescente necessidade da elaboração de substitutos de tecidos para o tratamento de feridos e doentes, assim como, há grande demanda de pele para testes químicos das áreas farmacêutica e cosmética. Outro fator de fundamental importância para o desenvolvimento de métodos alternativos in vitro, é a pressão mundial para que estes testes substituam os modelos animais. Esta abordagem vai de encontro aos novos conceitos de substituição, redução e refinamento na utilização de animais em estudos científicos, ditando o futuro da cultura celular e bioengenharia de tecidos. Graças ao grande desenvolvimento do cultivo celular e descoberta de que as células cultivadas podem ser reagrupadas de acordo com o delineamento experimental...

Efeito do composto natural Yo Jyo Hen Shi Ko (YHK) no ciclo de replicação do vírus da hepatite C (VHC); Effects of the natural compound Yo Jyo Hen Shi Ko (YHK) on the replication cycle of the hepatitis C virus

Pereira, Isabel Veloso Alves
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Tese de Doutorado Formato: application/pdf
Publicado em 29/10/2014 Português
Relevância na Pesquisa
65.7%
Estima-se que 170 milhões de pessoas no mundo estejam infectadas com o vírus da hepatite C (VHC), o que está altamente relacionado à ocorrência de hepatite crônica e carcinoma hepatocelular. A prevalência de esteatose hepática em doentes com hepatite C crônica é muito maior do que na população geral variando entre 40 a 75%. A associação entre a infecção pelo VHC e esteatose hepática é multifatorial. Duas formas de esteatose hepática são encontradas em pacientes com hepatite C crônica: esteatose metabólica (fatores de risco) e citopática relacionada ao genótipo 3a. Os lipídios são essenciais para o ciclo de replicação do VHC, eles podem exercer seu efeito em diferentes níveis como: grupos prostéticos em proteínas virais e/ou cofatores celulares na replicação de VHC, componentes especializados na estrutura do VHC onde ocorre a replicação ou como constituinte das partículas lipovirais. Trabalhos experimentais realizados anteriormente por nosso grupo relataram que a administração do composto natural Yo Jyo Hen Shi Ko (YHK) promove a inibição do desenvolvimento da esteatose, redução dos marcadores de estresse oxidativo, menor escore de inflamação, melhora nas concentrações de aminotransferases e diminuição da gordura visceral em um modelo animal de esteato-hepatite não alcoólica. A terapia padrão da hepatite C consiste em uma combinação de interferon peguilado alfa (PEG-IFN-alfa) que estimula o sistema imunológico do hospedeiro para combater a infecção e o composto antiviral ribavirina. Atualmente foram aprovados pelas agências de saúde os inibidores de protease Boceprevir...

Human breast tumor slices as an alternative approach to cell lines to individualize research for each patient

Conde, Sandro Jose; Melo Luvizotto, Renata de Azevedo; de Sibio, Maria Teresa; Nogueira, Celia Regina
Fonte: Lippincott Williams & Wilkins Publicador: Lippincott Williams & Wilkins
Tipo: Artigo de Revista Científica Formato: 333-335
Português
Relevância na Pesquisa
55.75%
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Processo FAPESP: 05/55459-1; There are several breast cancer experimental models including cell lines, which are commonly used due to ease of handling and storage. However, the continued propagation of cell lines and distribution among laboratories results in genetic drift and distancing from the in-vivo model. Primary organ culture of breast cancer slices may produce biological responses with high standard deviation for different samples, reflecting the heterogeneity of different tumors. Thus, the organ culture model system offers a new perspective to the results obtained in the cell lines and offers an alternative for studies that seek to individualize treatment for each patient, an increasingly prominent concern in current cancer therapy. European Journal of Cancer Prevention 21:333-335 (C) 2012 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

Root instrumentation with an erbium:yttrium-aluminum-garnet laser: Effect on the morphology of fibroblasts

Rossa Júnior, Carlos; Silvério, Karina G.; Zanin, Iriana C. J.; Brugnera Júnior, Aldo; Sampaio, José Eduardo C.
Fonte: Universidade Estadual Paulista Publicador: Universidade Estadual Paulista
Tipo: Artigo de Revista Científica Formato: 496-502
Português
Relevância na Pesquisa
55.74%
Objective: The purpose of this study was to evaluate the effect of erbium:yttrium-aluminum-garnet laser instrumentation of root surfaces on the morphology of fibroblasts from continuous lineage. Method and materials: Dentinal slices with 4 mm2 of surface area were obtained from teeth extracted for severe periodontal involvement. Specimens were assigned to one of three treatment groups: group 1, application of the laser with an energy level of 250 mJ at 103 pulses per second; group 2, application of the laser with an energy level of 80 mJ at 166 pulses per second; and group 3, similar to group 2, but with concomitant water irrigation of the device. The specimens were incubated in multiwell plates containing cell culture media. After 24 hours, the specimens were submitted to routine preparation for scanning electron microscopy. Three independent and blind examiners used photomicrographs to evaluate the morphology of the fibroblasts: 0 = without cells; 1 = flat cells; 2 = round cells; and 3 = combination of round and flat cells. Results: Statistical analysis indicated that there were significant differences among treatment groups and that group 3 was significantly different from groups 1 and 2. Conclusion: There was no difference between groups 1 and 2 in the morphology of fibroblasts. Laser instrumentation with concomitant irrigation impaired the adhesion of fibroblasts to dentinal surfaces.

Investigação citogenética em indivíduos com mosaico pigmentar do tipo Ito; Cytogenetic analysis in individuals with mosaic pigmentary of Ito type

Karina Soares Cunha
Fonte: Biblioteca Digital da Unicamp Publicador: Biblioteca Digital da Unicamp
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 26/08/2010 Português
Relevância na Pesquisa
55.8%
O Mosaico pigmentar tipo Ito é uma alteração cutânea frequente, caracterizada por hipopigmentação da pele que, na maioria dos casos, segue o padrão linhas de Blaschko, geralmente associada a anomalias extracutâneas, sobretudo anomalias do Sistema Nervoso Central (SNC). Sugere-se que esse padrão decorre da presença e migração de duas linhagens celulares no período embrionário, com diferente expressão de genes associados à pigmentação, que darão origem à epiderme e ao SNC no feto. Diversos tipos de mosaicismo foram associados ao quadro e acredita-se que os casos em que não houve tal detecção se devam à limitação das células analisadas. Devido à função, origem embrionária e migração celular, possivelmente o mosaicismo seria melhor identificado em melanócitos e queratinócitos, principalmente na presença de alterações no SNC, as alterações poderiam ter envolvimento com o prognóstico neurológico. Neste estudo foi realizada análise citogenética de linfócitos e fibroblastos de 15 indivíduos com mosaico pigmentar do tipo Ito. Os objetivos incluíram padronizar a cultura de células de melanócitos e queratinócitos, visando analisar o cariótipo desses indivíduos nesses tipos celulares. O estudo citogenético nesses tipos celulares...

Establishing a stem cell culture laboratory for clinical trials

Sekiya,Elíseo Joji; Forte,Andresa; Kühn,Telma Ingrid Borges de Bellis; Janz,Felipe; Bydlowski,Sérgio Paulo; Alves,Adelson
Fonte: Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular Publicador: Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/01/2012 Português
Relevância na Pesquisa
85.84%
Adult stem/progenitor cells are found in different human tissues. An in vitro cell culture is needed for their isolation or for their expansion when they are not available in a sufficient quantity to regenerate damaged organs and tissues. The level of complexity of these new technologies requires adequate facilities, qualified personnel with experience in cell culture techniques, assessment of quality and clear protocols for cell production. The rules for the implementation of cell therapy centers involve national and international standards of good manufacturing practices. However, such standards are not uniform, reflecting the diversity of technical and scientific development. Here standards from the United States, the European Union and Brazil are analyzed. Moreover, practical solutions encountered for the implementation of a cell therapy center appropriate for the preparation and supply of cultured cells for clinical studies are described. Development stages involved the planning and preparation of the project, the construction of the facility, standardization of laboratory procedures and development of systems to prevent cross contamination. Combining the theoretical knowledge of research centers involved in the study of cells with the practical experience of blood therapy services that manage structures for cell transplantation is presented as the best potential for synergy to meet the demands to implement cell therapy centers.

Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR

Vinatea,Cecília E.B.; Sincero,Thaís C.M.; Simões,Claúdia M.O.; Barardi,Célia R.M.
Fonte: Sociedade Brasileira de Microbiologia Publicador: Sociedade Brasileira de Microbiologia
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/03/2006 Português
Relevância na Pesquisa
65.72%
Shellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses are generally used as a model for the detection of viruses from shellfish due to their public health significance. In the present work, oysters were placed in glass aquaria containing seawater plus unicellular algae. Two experiments were performed: 1) oysters bioaccumulating four different poliovirus type 2 concentrations: 5 x 10(4), 2.5 x 10(4), 5 x 10³ and 5 x 10² PFU/mL during 20h; 2) oyster tissues directly inoculated with 6.0 x 10(5) and 1.0 x 10(5) PFU/mL. After viruses seeding, tissue samples were processed by an adsorption-elution-precipitation method. Positive controls were performed by seeding 6.0 x 10(5) PFU/mL of poliovirus type 2 directly on the final oyster tissue extracts. Oyster extracts were assayed for viruses recovery by plaque assay, RT-PCR and integrated cell culture-PCR methodologies (ICC/PCR). The last one was based on the inoculation of the samples onto VERO cell monolayer followed by RT-PCR analysis of the infected cell fluid. In the first experiment (20h bioaccumulation) until 5 x 10³ PFU were detected after 24 and 48h growth on VERO cells. Direct RT-PCR and ICC/PCR were able to detect 3 and 0.04 PFU of poliovirus...

Fundamentals of microfluidic cell culture in controlled microenvironments†

Young, Edmond W. K.; Beebe, David J.
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
55.89%
Microfluidics has the potential to revolutionize the way we approach cell biology research. The dimensions of microfluidic channels are well suited to the physical scale of biological cells, and the many advantages of microfluidics make it an attractive platform for new techniques in biology. One of the key benefits of microfluidics for basic biology is the ability to control parameters of the cell microenvironment at relevant length and time scales. Considerable progress has been made in the design and use of novel microfluidic devices for culturing cells and for subsequent treatment and analysis. With the recent pace of scientific discovery, it is becoming increasingly important to evaluate existing tools and techniques, and to synthesize fundamental concepts that would further improve the efficiency of biological research at the microscale. This tutorial review integrates fundamental principles from cell biology and local microenvironments with cell culture techniques and concepts in microfluidics. Culturing cells in microscale environments requires knowledge of multiple disciplines including physics, biochemistry, and engineering. We discuss basic concepts related to the physical and biochemical microenvironments of the cell, physicochemical properties of that microenvironment...

Advanced Cell Culture Techniques for Cancer Drug Discovery

Lovitt, Carrie J.; Shelper, Todd B.; Avery, Vicky M.
Fonte: MDPI Publicador: MDPI
Tipo: Artigo de Revista Científica
Publicado em 30/05/2014 Português
Relevância na Pesquisa
55.75%
Human cancer cell lines are an integral part of drug discovery practices. However, modeling the complexity of cancer utilizing these cell lines on standard plastic substrata, does not accurately represent the tumor microenvironment. Research into developing advanced tumor cell culture models in a three-dimensional (3D) architecture that more prescisely characterizes the disease state have been undertaken by a number of laboratories around the world. These 3D cell culture models are particularly beneficial for investigating mechanistic processes and drug resistance in tumor cells. In addition, a range of molecular mechanisms deconstructed by studying cancer cells in 3D models suggest that tumor cells cultured in two-dimensional monolayer conditions do not respond to cancer therapeutics/compounds in a similar manner. Recent studies have demonstrated the potential of utilizing 3D cell culture models in drug discovery programs; however, it is evident that further research is required for the development of more complex models that incorporate the majority of the cellular and physical properties of a tumor.

Avaliação in vitro da adesão e proliferação de células mesenquimais do ligamento periodontal humano em diferentes superfícies de titânio

Ribeiro, Rodrigo Alves
Fonte: Universidade Federal do Rio Grande do Norte; BR; UFRN; Programa de Pós-Graduação em Odontologia; Odontologia Preventiva e Social; Periodontia e Prótese Dentária Publicador: Universidade Federal do Rio Grande do Norte; BR; UFRN; Programa de Pós-Graduação em Odontologia; Odontologia Preventiva e Social; Periodontia e Prótese Dentária
Tipo: Dissertação Formato: application/pdf
Português
Relevância na Pesquisa
65.79%
In the last years, many scientific researches in implantology have been focused on alternatives that would provide higher speed and quality in the process of osseointegration. Different treatment methods can be used to modify the topographic and chemical properties of titanium surface in order to optimize the tissue-implant reactions by a positive tissue response. This study aimed to evaluate the adhesion and proliferation of mesenchymal cells from human periodontal ligament on two different titanium surfaces, using cell culture techniques. Grade II titanium discs received different surface treatments, forming two distinct groups: polished and cathodic cage plasma nitriding. Human periodontal ligament mesenchymal cells were cultured on titanium discs in 24-well cell culture plates, at a density of 2 x 104 cells per well, including wells with no discs as positive control. Data obtained by counting the cells that adhered to the titanium surfaces (polished group and cathodic cage group) and to the plastic surface (control group), in the 24, 48 and 72-hour periods after plating, were used to analyze cell adhesion and proliferation and to obtain the cell growing curve in the different groups. The data were submitted to nonparametric analysis and the differences between groups were compared by Kruskal-Wallis and Friedman statistical tests. No statistically significant differences were found in the cells counts between the groups (p>0.05). It was concluded that both treatments produced surfaces compatible with the adhesion and proliferation of human periodontal ligament mesenchymal cells; Nos últimos anos...

Culture without the petri-dish

Thompson, J.
Fonte: Elsevier Science Inc Publicador: Elsevier Science Inc
Tipo: Artigo de Revista Científica
Publicado em //2007 Português
Relevância na Pesquisa
55.77%
Automation of oocyte maturation and embryo production techniques is a new and exciting development in the field of reproductive technologies. There are two areas where increased automation is having an impact: in the area of embryo diagnostics and in the process of embryo production itself. Benefits include decreased staffing and skill requirements for production and assessment of embryos, as well as increasing quality management systems by removing the “human” factor. However, the uptake of new technologies is likely to be slow, as costs and the conservative nature of the Assisted Reproduction Technology industry to adopt new techniques.; Jeremy G. Thompson; Available online 20 October 2006.

Biotechnology Apprenticeship for Secondary-Level Students: Teaching Advanced Cell Culture Techniques for Research

Lewis, Jennifer R.; Kotur, Mark S.; Butt, Omar; Kulcarni, Sumant; Riley, Alyssa A.; Ferrell, Nick; Sullivan, Kathryn D.; Ferrari, Mauro
Fonte: American Society for Cell Biology Publicador: American Society for Cell Biology
Tipo: Artigo de Revista Científica
Publicado em //2002 Português
Relevância na Pesquisa
75.9%
The purpose of this article is to discuss small-group apprenticeships (SGAs) as a method to instruct cell culture techniques to high school participants. The study aimed to teach cell culture practices and to introduce advanced imaging techniques to solve various biomedical engineering problems. Participants designed and completed experiments using both flow cytometry and laser scanning cytometry during the 1-month summer apprenticeship. In addition to effectively and efficiently teaching cell biology laboratory techniques, this course design provided an opportunity for research training, career exploration, and mentoring. Students participated in active research projects, working with a skilled interdisciplinary team of researchers in a large research institution with access to state-of-the-art instrumentation. The instructors, composed of graduate students, laboratory managers, and principal investigators, worked well together to present a real and worthwhile research experience. The students enjoyed learning cell culture techniques while contributing to active research projects. The institution's researchers were equally enthusiastic to instruct and serve as mentors. In this article, we clarify and illuminate the value of small-group laboratory apprenticeships to the institution and the students by presenting the results and experiences of seven middle and high school participants and their instructors.

Replacement of fetal calf serum by human serum as supplementation for human fibroblast culture

Isaac,César; Mattos,Cristiana Nicoli de; Rêgo,Francinni Mambrine Pires do; Cardim,Larissa Nocchi; Altran,Silvana Cereijido; Paggiaro,André Oliveira; Tutihashi,Rafael Mamoru Carneiro; Mathor,Mônica Beatriz; Ferreira,Marcus Castro
Fonte: Sociedade Brasileira de Cirurgia Plástica Publicador: Sociedade Brasileira de Cirurgia Plástica
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/09/2011 Português
Relevância na Pesquisa
65.78%
INTRODUCTION: Fetal calf serum (FCS) is commonly used as a supplement in the culture medium for fibroblast cells. This supplementation is far from ideal as sample quality varies from batch to batch and the composition of FCS is not completely known. In addition, FCS may be contaminated with viruses and/or prions and may also cause adverse immunologic responses in humans. Due to these facts, a worldwide effort is being made to find alternatives for xenobiotic elements in cell cultures. Human serum could be a safer alternative, especially for clinical application. METHODS: We investigated human serum as a substitute for FCS in human fibroblast culture. Fresh human serum was obtained from 10 healthy volunteers. Fibroblasts were cultivated in multiwell plates containing either Dulbecco's modified Eagle's medium (DMEM) plus 10% FCS (D10) or DMEM plus 10% human serum (D10H). Cell counts were obtained between 24 and 264 hours of cultivation; results were expressed as the mean number of cells ± standard error of the mean to create cell proliferation curves. RESULTS: There was no statistical difference in fibroblast proliferation between the two groups. Human serum supported human fibroblast growth and proliferation, suggesting that it may be a potential substitute for FCS in human cell culture. Cells cultivated with human serum presented a different morphology...

Comparative study of two different methods of harvesting adipose tissue derived stem cell; Estudo comparativo de dois métodos para obtenção de células mesenquimais indiferenciadas de tecido adiposo

Rêgo, Francinni Mambrini Pires; Cruz, Aline de Paula da; Mattos, Cristina Nicoli de; Tutihashi, Rafael Mamoru Carneiro; Isaac, César; Ferreira, Marcus Castro
Fonte: Universidade de São Paulo. Faculdade de Medicina Publicador: Universidade de São Paulo. Faculdade de Medicina
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; Formato: application/pdf
Publicado em 06/03/2009 Português
Relevância na Pesquisa
65.73%
Introduction: Mesenchymal tissue loose can lead to large wounds, which treatmentsare not completely free of morbidity. The adipose tissue derived stem cell participation in woundhealing process has been widely studied. Objective: The aim of the study is to compare twodifferent methods of harvesting adipose tissue derived stem cell. Methods: Standard amountsof adipose tissue from lipectomy (L) and liposuction (LA) have been kept in cell culture for 72hours in 6 well clusters and then discharged. 96 hours after the discharge cells were tripsynizedfrom the multi-well culture surface and counted on hemocitometer chamber. Results: The meannumber of cells from LA group was 9.4 x 104, in the L group the number of cell was 0 (zero).Conclusion: Comparing the 2 methods, cells were harvested only from the liposuction group.; Introdução: Perdas de tecido mesenquimal podem acarretar grandes áreascruentas, cujos tratamentos nem sempre são isentos de morbidade. A participação de célulasmesenquimais indiferenciadas, no processo de cura de feridas, tem sido amplamente estudada.Objetivo: comparar dois métodos de obtenção de células mesenquimais indiferenciadasde tecido adiposo. Métodos: amostras padronizadas de fragmentos de tecidos gordurososprovenientes de lipectomia (L) e de lipoaspiração (LA) foram mantidas em cultura celular emplacas multipoços de 6 poços por 72 horas e posteriormente descartadas. Após 96 horas dodescarte...

Cultivo de células-tronco derivadas de tecido adiposo: uma análise crítica; Culture of adipose-derived stem cells: a critical analysis

Ladeira, Pedro Ribeiro Soares de; Isaac, Cesar; Nakamura, Yeda Midori; Tutihashi, Rafael Mamoru Carneiro; Paggiaro, André Oliveira; Ferreira, Marcus Castro
Fonte: Universidade de São Paulo. Faculdade de Medicina Publicador: Universidade de São Paulo. Faculdade de Medicina
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; Formato: application/pdf
Publicado em 18/12/2012 Português
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Introduction: adipose-derived stem cells (ADSCs) are pluripotent and present little invasive collection and high cell yield in culture, besides the fact that the number of liposuctions, the main source of these cells, is increasing around the world. Plastic Surgery has found the benefits of the use of those cells in several areas, which increases the importance of critically analyzing the protocols currently used for cell cultivation. Objective: to describe in detail the cultivation protocol adopted by our laboratory in order to perform a critical analysis of the culture of ADSCs. Method: surpluses of adipose tissue were donated to the laboratory with research purposes for 5 patients of both genders and age between 20 and 45 years. From each patient, 30 mL of fatty material was collected in a sterile flask and transported to the laboratory for cell culture, where they were subjected during 45 min to enzymatic digestion by 30 mg of type IA collagenase diluted in 30mL of Dulbecco’s Modified Eagle Medium (DMEM) and then centrifuged to isolate the cells from the extracellular matrix. The amplification of the cultures was made with 0.05% trypsin solution + EDTA 0.02%. For cryopreservation was used freezing media consisting of 60% DMEM...

; Comparative isolation protocols and characterization of stem cells from human primary and permanent teeth pulp

Souza, Leliane Macedo de; Bittar, Juliana Duarte; Silva, Izabel Cristina Rodrigues da; Toledo, Orlando Ayrton de; Brígido, Marcelo de Macedo; Fonseca, Marcio José Poças
Fonte: UNICAMP/FOP Publicador: UNICAMP/FOP
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; ; ; Formato: application/pdf
Publicado em 16/11/2015 Português
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; Aim: This study was developed to compare the morphological, proliferative and immunophenotypic profiles of pulp cells from permanent and primary teeth, obtained by two isolation methods. Methods: Normal human impacted third molars and exfoliated primary teeth were collected and cut around the cementoenamel junction. Pulp cells cultures were established by two approaches: enzyme digestion (3 mg/mL type I colagenase and 4 mg/mL dispase), or culture of the tissue explants in cell culture dishes. Morphological and proliferative analyses, as well as immunophenotype characterization with monoclonal antibodies against CD117, CD34 and CD45 surface receptors were performed. Results: For the permanent teeth, on the 4th day of culture, the cell number was significantly higher for the outgrowth method. By the end of the studied period (14th day), the enzymatic method was more efficient in promoting culture growth. On the other hand, for primary teeth, enzymatic digestion always promoted a higher cell proliferation. The immunophenotypic profiles were CD117+/ CD34-/ CD45- and CD117+/ CD34+/ CD45- for cells from permanent and primary teeth, respectively. Conclusions: The findings of this study indicate that both isolation methods can be efficiently used. The cell population displayed an immunophenotype compatible to the one of stem cells...