Influência do índice apoptótico e da imuno-expressão da survivina no prognóstico de pacientes com mola hidatiforme completa

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Determinação da ativação de monócitos e de biomarcadores celulares em gestantes portadoras de pré-eclâmpsia

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Papel de neutrófilos humanos no direcionamento da resposta imune adaptativa do tipo Th1 contra o Paracoccidioides brasiliensis

Pós-graduação em Doenças Tropicais - FMB

Foto seleção de mielomas e hibridomas murinos na produção de anticorpos monoclonais

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Do progenitor cells from different tissue have the same phenotype?

The purpose of this work was to validate isolation methods for sheep mesenchymal stem cells (MSC) from different sources and to explore the hypothesis that MSC exhibit markers of the same phenotype independent from tissue source. Cells derived from ovine bone marrow, synovial membrane and adipose tissue were characterized using the following markers: CD44, CD45, CD11b and MHC-I. The isolated MSC were cultivated, went through osteogenic, chondrogenic and adipogenic differentiation, and were characterized by flow cytometry using mouse anti-ovine CD44, CD45 and MHC-I monoclonal antibody (mAb), and mouse anti-bovine CD11b mAb. Ovine MSC from all three sources differentiated under chondorgenic, osteogenic and adipogenic conditions. Also, MSC from the three tissues were found to express CD44 and MHC-I but lack of CD11b and CD45. The results obtained revealed that our isolation methods for the different tissues tested are valid and that MSC from the three sources studied have same immunophenotic characteristics. (C) 2014 Published by Elsevier Ltd. All rights reserved.

Leishmania amazonensis Promastigotes Present Two Distinct Modes of Nucleus and Kinetoplast Segregation during Cell Cycle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immunohistochemical localization of nicotinic acetylcholine-receptor subunits in the mesencephalon and diencephalon of the chick (Gallus-gallus)

Monoclonal antibodies against two alpha-bungarotoxin-binding subunits (alpha-7 and alpha-8) of the nicotinic acetylcholine receptors (nAChRs) were used as immunohistochemical probes to map their distribution in the chick diencephalon and mesencephalon. The distribution of the alpha-7 and alpha-8 nAChR subunits was compared to the distribution of immunoreactivity produced by a monoclonal antibody against the beta-2 structural subunit of the nAChRs.Structures that contained high numbers of alpha-7-like immunoreactive (LI) somata included the intergeniculate leaflet, nucleus intercalatus thalami, nucleus ovoidalis, organum paraventricularis, nucleus rotundus, isthmic nuclei, nucleus trochlearis, oculomotor complex, nucleus interstitio-pretecto-subpretectalis, stratum griseum centrale of the optic tectum, and nucleus semilunaris. Neuropil staining for alpha-7-LI was intense in the nucleus dorsomedialis hypothalami, nucleus geniculatus lateralis ventralis, griseum tecti, isthmic nuclei, nucleus lentiformis mesencephali, nucleus of the basal optic root, and stratum griseum et fibrosum superficiale of the tectum. High numbers of alpha-8-LI somata were found in the stratum griseum et fibrosum superficiale of the tectum and the nucleus interstitio-pretecto-subpretectalis, and intense neuropil staining for alpha-8-LI was found in the dorsal thalamus, nucleus geniculatus lateralis ventralis, lateral hypothalamus, griseum tecti, nucleus lentiformis mesencephali, nucleus interpeduncularis, and stratum griseum et fibrosum superficiale of the tectum. High numbers of beta-2-LI somata were found only in the nucleus spiriformis lateralis, whereas neuropil staining for beta-2-LI was intense in the nucleus geniculatus lateralis ventralis, nucleus suprachiasmaticus, nucleus lateralis anterior, nucleus habenularis lateralis, area pretectalis, griseum tecti, nucleus lentiformis mesencephali, nucleus externus, and nucleus interpeduncularis, and in the stratum griseum centrale, stratum griseum et fibrosum superficiale, and stratum opticum of the tectum.These results indicate that there are major disparities in the localization of the alpha-bungarotoxin-binding alpha-7 and alpha-8 nAChR subunits and the beta-2 structural nAChR subunit in the chick diencephalon and mesencephalon. These nAChR subunits appear, however, to coexist in several regions of the chick brain.

Interferon-gamma production by human neutrophils upon stimulation by IL-12, IL-15 and IL-18 and challenge with Paracoccidioides brasiliensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Produção de anticorpos monoclonais murinos dirigidos contra células-tronco mesenquimais de coelho

Mesenchymal stem cells (MSCs) are a heterogeneous population of cells that proliferate in vitro as plastic-adherent cells, have fibroblast-like morphology and can differentiate into bone, cartilage and fat cells. Therapeutic potential of MSCs have been studied in experimental models, such as rabbit, in Laboratory of Cell Engineering of Botucatu. However, no specific markers have been reported for expanded rabbit MSCs, which hampers the isolation of pure MSC populations by immunophenotypic characterization. Thus, the objective of this study was to produce monoclonal antibodies (mAbs) to rabbit MSCs. MSCs derived from rabbit bone marrow (BM) were isolated, cultured, expanded ex vivo, and immunized into three BALB/c mices, and spleen cells subsequently harvested were used to generate hibridoma cell lines secreting antibodies against MSCs. Hybridoma cells were screened by flow cytometry and antibody-producing cells were subjected to subsequent rounds of retests. MSC1-160 obtained the best positivity for IgG expression and was cloned by limiting dilutions and micromanipulation. Ascitic fluid from ten best clones was purified by affinity chromatography in Protein A-sepharose CL-4B column and purification control was performed by electrophoresis in agarose gels. The purified IgG were tested against rabbit MSCs, obtaining high positivity by flow Cytometry. In conclusion, we developed 10 mAbs, MSC1-160 A20, A30, A41, A47, A55, A60, A63, A69, A81, and A82, that recognize rabbit MSC cell surface antigens showing potential for immunophenotypic characterization of rabbit MSC cell lines

Análise da expressão de antígenos câncer/testítulo (CTs) em pacientes com mieloma múltiplo

O mieloma múltiplo (MM) é uma neoplasia maligna secundária à expansão clonal de células plasmáticas caracterizada pela presença de imunoglobulina monoclonal no sangue e/ou na urina, lesões líticas ósseas e infiltração de plasmócitos monoclonais na medula óssea. A caracterização dos mecanismos responsáveis pela expansão das células tumorais do MM é difícil e envolve uma série de alterações genéticas e mudanças no microambiente da medula óssea, favorecendo o crescimento do tumor e a falha do sistema imune em reconhecê-lo. O MM é uma doença incurável, sendo a sobrevida mediana dos pacientes em torno de 3-5 anos. Terapias atuais como quimioterapia e transplante autólogo de células-tronco podem induzir remissão da doença, mas a recaída e a morte são inevitáveis. Os antígenos específicos câncer/testículo (CTs) foram originalmente descritos em pacientes com melanoma e são assim denominados, pois suas proteínas foram identificadas em espermatogônias, células conhecidas por não expressarem antígenos de histocompatibilidade (HLA), o que as impossibilita de desencadear uma resposta imune específica. Esses antígenos foram identificados posteriormente em vários tipos de tumores humanos, como melanoma, carcinoma pulmonar, câncer renal, entre outros. Até o momento, existem poucas informações a respeito de sua importância como fatores de prognóstico clínico ou relacionado à proliferação aberrante das células plasmáticas. Assim, o objetivo deste estudo é avaliar o nível de expressão gênica dos antígenos câncer/testículo MAGEC1(CT7), MAGEA3/6, LAGE-1, NY-ESO-1 e GAGE em amostras de aspirado de medula óssea total de pacientes com mieloma múltiplo. A análise de expressão por RT-PCR dos 5 CTs de interesse, realizada em 21 amostras de medula total de pacientes com MM, demonstrou as seguintes freqüências: CT7 (42,9%), LAGE-1 (23,8%)... (Resumo completo, clicar acesso eletrônico abaixo)

Marcadores de superfície de Células-tronco: estudo de reação cruzada inter-espécies

Mesenchymal stem cells (MSCs) are adult multipotent cells with fibroblastoid morphology and adherent to plastic. Furthermore, they can be obtained from different sources. Besides bone marrow, these cells are taken from umbilical cord blood, umbilical vein, saphenous vein, peripheral blood, arteries, liver and fetal pancreas, placenta, dental pulp and adipose tissue. MSCs derived from adipose tissue are important because of the abundant number of cells that can be obtained from this tissue, easy access and little discomfort to the patient. This study compared two techniques for obtaining MSCs from adipose tissue: mechanical dissociation (MD) and enzymatic digestion (ED). We also analyzed the inter-species cross-reactions using commercial monoclonal antibodies directed against surface antigens of stem cells from different species: mouse, horse, rabbit, monkey and human. We found that MD technique is favorable in relation to ED within 15 days of culture, and ED is more efficient in the first days of culture. The data also showed that MD causes less damage to cellular DNA. About inter-species cross-reactions, the monoclonal antibody A69 directed against stem cells from rabbits, which can be used in veterinary medicine, particularly in research involving horses

Influência da luz LED na cultura celular de Mieloma múltiplo murino–NS1 in vitro

A monoclonal antibody (mAb) is an important tool in medical biotechnology and the production of biopharmaceuticals, especially for disease diagnosis and treatment of infections, because the antibodies have a significant advantage over chemical agents used in conventional therapies . The last thirty years the technology of production of monoclonal antibodies developed mainly the technique of obtaining in vitro, but also of their production is laborious, the cost is high. A major element of the high cost of production is the fact that the long-term culture consumes a large amount of imported inputs with high added value. A major contribution of this work is to promote cell growth more quickly and efficiently. Currently, a great race to discover new technologies and techniques to synthesize new antibodies and significantly increase the production of murine mAbs. New technologies such as laser and LED are innovations and widespread in modern life, so much so that its use has proliferated worldwide, primarily in the medical field. Recent studies show a series of results from the influence of the LED light in biological tissues such as: increasing the rate of cell proliferation, increased production rate of fibroblasts, increasing the rate of synthesis of RNA and DNA synthesis of ATP, etc. To assess the contribution of the LED in the culture of Myeloma NS1murino compared to the standard procedure. - NS1 cells were provided and followed the criteria of culture medium of the Laboratory of Cellular Engineering Center of Botucatu (POPs). The same amount of cells was grown in bottles of 25 cm2 polystyrene Tissue Culture Treated, specifically marked and kept in special medium RPMI 1640 Gibco BRL  supplemented with fetal bovine serum 10%, essential amino acids and non-essential, glucose, insulin and antibiotics. It was used in LEDs Cromatek wavelength of 630nm, 475nm and 530nm. The groups were... (Complete abstract click electronic access below)

Avaliação da apoptose e resposta Th17 em modelo murino de esporotricose

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Análise funcional da proteína 14-3-3 de Paracoccidioides brasiliensis e prospecção de alvos terapêuticos inibidores da interação de P. brasiliensis à pneumócitos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)