Browsing by Subject "Cell culture"

Now showing 1 - 9 of 9
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    Caracterización de exosomas producidos por células oviductales in vivo e in vitro, en la especie bovina
    (Facultad de Veterinaria y el Servicio de Publicaciones de la Universidad de Murcia, 2023) Toledo Guardiola, Santa María; Matás Parra, Carmen; Rueda Gomariz, Almudena
    Las vesículas extracelulares (VEs), exosomas y micro vesículas son un tipo de estructuras heterogéneas pre-sentes en la mayoría de los fluidos orgánicos incluyendo el fluido oviductal. Las VEs contienen varios compuestos derivados de la célula original, como proteínas, lípidos, ARNm, miARN y ADN. Las VEs en el oviducto son producidas por las células epiteliales y entre sus funciones se encuentran: la interacción con los espermatozoides, mantener la viabilidad de éstos, participar en la maduración de los ovocitos y en el proceso de fecundación. Durante la fecundación in vitro y con el fin de mejorarla imitando las condiciones in vivo, numerosos investigadores han utilizado cultivos de células del epitelio oviductal bovino (CEOB) con notables mejoras. Estas células producen, entre otros componentes VEs, por ello, en este trabajo hemos planteado un estudio com-parativo de VEs presentes en el fluido oviductal (FO) bovino recogido en momentos próximos a la ovulación (in vivo) y de aquellas VEs producidas en cultivos de CEOB a los 7 días de cultivo (in vitro) comparando el tamaño, la distribución de la población y la concentración de proteína en ambos tipos. Las VEs se identificaron mediante microscopía electrónica, su tamaño mediante dispersión de luz láser y la concentración de proteínas mediante el método Bradford. Los resultados mostraron que el tamaño de las VEs fue similar entre ambos grupos experimentales. Por otro lado, sí que se observaron diferencias en cuanto a la concentración de proteínas. Las VEs obtenidas in vivocontenían mayor cantidad de proteína en su cargo que en las VEs obtenidas in vitro.En cuanto a identificación de las VEs mediante microscopía electrónica de transmisión, solo pudieron ser observadas aquellas obtenidas in vivo. Este hecho podría deberse al lugar de dónde han sido recogidas, al mét-odo de cultivo de células epiteliales oviductales bovinas o la escasez en su producción.
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    Characterisationof thyroid medullary carcinoma TT cell line
    (Murcia : F. Hernández, 1997) Zabel, M.; Grzeszkowak, J.
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    Development of betalain producing callus lines from colored quinoa varieties (Chenopodium quinoa Willd)
    (American Chemical Society, 2017-12-14) Henarejos Escudero, Paula; Guadarrama-Flores, Berenice; Guerrero Rubio, María Alejandra; Gómez-Pando, Luz Rayda; Gandía Herrero, Fernando; García Carmona, Francisco; Bioquímica y Biología Molecular "A"
    Betalains are water-soluble plant pigments of hydrophilic nature with promising bioactive potential. Among the scarce edible sources of betalains is the grain crop quinoa (Chenopodium quinoa Willd), with violet, red, and yellow grains being colored by these pigments. In this work, call us cultures have been developed from differently colored plant varieties. Stable callus lines exhibited color and pigment production when maintained on Murashige and Skoog medium supplemented with the plant growth regulators 6-benzylaminopurine (8.88 μM) and 2,4-dichlorophenoxyacetic acid (6.79 μM) with a reduction of the nitrogen source to 5.91 mM. Pigment analysis by HPLC-DAD and ESI-MS/MS fully describes the content of individual pigments in the cell lines and allows the first report on the pigments present in quinoa seedlings. Phyllocactin and vulgaxanthin Iare described as novel pigments in the species and show the potential of C. quinoa culture lines in the production of compounds of nutritional value.
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    Early human trophoblast cell cultures. A morphological and immunocytochemical study
    (Murcia : F. Hernández, 1989) Logothetou-Rella, H.; Kotoulas, I-G.; Nesland, Jahn M.; Kipiotis, D.; Abazis, D.
    Rapidly growing cytotrophoblasts were isolated from early human chorionic villi and the Papanicolaou method was used to characterize their cytology and transformation into syncytiotrophoblasts. Cytotrophoblasts fused and formed binucleated cells or mononucleated intermediate cells. Syncytial cells were formed by fusion of small cytotrophoblasts or intermediate cells and cytotrophoblasts. Glycosaminoglycans were produced in cytotrophoblasts and released extracellularly . Here they were accumulated andlor diffused into a continuous layer covering the cells. Glycosaminoglycans in syncytial cells were contained in well defined membranous sacs. Cytotrophoblasts only grown beyond confluence differentiated into villi with a villus-like histology.
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    Electron microscopic observations on the formation of elastic fibres in cultures of aortic medial cells and adventitial cells
    (Murcia : F. Hernández, 1988) Masumi Akita; Katsuji Kaneko; Merker, H. J.
    The formation of elastic fibres was observed in the cultured cells derived from the tunica media and the tunica adventitia of mouse aorta. Bundles of myofilaments with dense bodies were abundantly observed in the cytoplasm of the cultured media1 cells, and numerous bundles of microfibrillar components were present in the intercellular spaces. Fine granules of approximately 50 nm in diameter were observed in the bundles of microfibrillar components. lt was supposed that these fine granules of elastin fused with each other and formed elastic aggregates and then formed large elastic clumps. Numerous bundles of microfibrillar components were also present in the intercellular spaces of the cultured adventitial cells. Elastic aggregates were scarcely observed in the bundles of microfibrillar components. However, large elastic clumps as observed in the media1 cell culture could not be found in the adventitial cell culture. It is suggested that the formation of large elastic clumps might be related to the sheet structures or lamellae of elastic fibres in the tunica media.
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    Mitochondrial metabolism characterization of four different fish cell lines
    Espinosa-Ruiz, Cristóbal; Mayor-Lafuente, Javier; Esteban Abad, María de los Ángeles; Biología Celular e Histología
    Different cell lines (SAF-1, DLB-1, PLHC-1 and FuB-1) mitro stress (OCR and ECAR) raw data.
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    Rosiglitazone promotes the differentiation of Langerhans cells and inhibits that of other dendritic cell types from CD133 positive hematopoietic precursors
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Bonetti, Maria Ida; Bacci, Stefano; Santosuosso, Michela; Mazzanti, Benedetta; Aldinucci, Alessandra; Ballerini, Clara; Guasti, Daniele; Calosi, Laura; Bosi, Alberto; Romagnoli, Paolo
    Dendritic cells and their precursors express PPAR-gamma, whose stimulation has inhibitory effects on the maturation and function of dendritic cells in vivo. Dendritic cells can differentiate in vitro from CD133+ progenitors; the influence of PPAR-gamma stimulation on this process is unknown. We have addressed the effect of PPAR-gamma agonist rosiglitazone, at a concentration as used in clinics, on the differentiation of dendritic cells from human CD133+ progenitors. Cells were harvested from cord blood by density gradient and immunomagnetic separation, and cultured for 18 days with fetal calf serum, cytokines and 1 µmol/L rosiglitazone. Analyses included flow cytometry, electron microscopy and mixed lymphocyte reaction. As expected, control cells generated without rosiglitazone were dendritic, expressed MHC-II, CD80, CD83 and CD86 and stimulated mixed reaction potently. A minority of cells expressed the Langerhans cell marker CD207/langerin, but none contained Birbeck granules. With rosiglitazone much fewer cells were generated; they were all dendritic, expressed differentiation and maturation-related antigens in higher percentage and were better stimulators of lymphocytes than those generated without the drug. The vast majority of cells expressed CD207/langerin and many contained Birbeck granules, i.e. were full-fledged Langerhans cells. We conclude that stimulation of PPAR-gamma, while negatively affecting the number of generated cells, promotes the maturation of human cord blood CD133 positive precursors into efficient, immunostimulating dendritic cells with a Langerhans cell phenotype.
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    The histopathology of a human mesenchymal stem cell experimental tumor model: support for an hMSC origin for Ewing`s sarcoma?
    (Murcia : F. Hernández, 2008) Burns, Jorge S.; Abdallah, Basem M.; Schröder, Henrik E.; Kassem, Moustapha
    Sarcomas display varied degrees of karyotypic abnormality, vascularity and mesenchymal with a branching periodic acid Schiff reaction pattern. Such clone-specific differences in host vascular response provide novel models to explore interactions between mesenchymal stem and endothelial cells. Despite the lack of a characteristic chromosomal translocation, the histomorphology, biomarkers and oncogenic changes were similar to those prevalent for Ewing’s sarcomas. The phenotype and ontogenesis of hMSC-TERT20 tumors was consistent with the hypothesis that sarcomas may arise from hMSC, providing a unique diploid model for exploring human sarcoma biology. differentiation. We have reported that a strain of telomerized adult human bone marrow mesenchymal stem cells (hMSC-TERT20) spontaneously evolved a tumorigenic phenotype after long-term continuous culture. We asked to what extent our hMSC-TERT20 derived tumors reflected events found in human sarcomas using routine histopathological procedures. Early versus late passage hMSC-TERT20 cultures persistently expressed mesenchymal lineage proteins e.g. CD105, CD44, CD99 and vimentin. However, late passage cultures, showed increased immunohistochemical staining for CyclinD1 and p21WAF1/Cip1, whereas p27Kip1 staining was reduced. Notably, spectral karyotyping showed that tumorigenic hMSC-TERT20 cells retained a normal diploid karyotype, with no detectable chromosome abnormalities. Consistent with the bone-forming potential of early passage hMSC-TERT20 cells, tumors derived from late passage cells expressed early biomarkers of osteogenesis. However, hMSC-TERT20 cells were heterogeneous for alpha smooth muscle actin (ASMA) expression and one out of six hMSC-TERT20 derived single cell clones was strongly ASMA positive. Tumors from this ASMA+ clone had distinctive vascular qualities with hot spots of high CD34+ murine endothelial cell density, together with CD34- regions
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    Ultrastructure of myotendinous junctions in tendon-skeletal muscle constructs engineered in vitro
    (Murcia: F. Hernández, 2009) Kostrominova, Tatiana Y.; Calve, Sarah; Arruda, Hellen M.; Larkin, Lisa M.
    During development, the interaction between tenocytes and myotubes leads to the formation of highly specialized muscle-tendon structural interfaces: myotendinous junctions (MTJs). Structural integrity of MTJs is critical for force transmission from contracting muscle through tendon to bone. We recently developed an in vitro model of three-dimensional (3-D) skeletal muscle-tendon constructs to address mechanisms of the MTJs development. We hypothesized that engineered in vitro 3-D skeletal muscle-tendon constructs would develop MTJs ultrastructurally resembling those found during fetal development in vivo. To test this hypothesis we compared MTJs structures in vivo to those developed in 3-D skeletal muscle constructs co-cultured with engineered self-organized tendon constructs (SOT), or segments of adult (ART) or fetal rat tail (FRT) by means of electron microscopy. Our study showed that at sites of termination some of the myofibers of the engineered 3-D skeletal muscle-FRT and -SOT constructs displayed emerging finger-like sarcolemmal projections surrounded by collagen fibers. These structures resemble fetal MTJs in vivo. Muscle-ART constructs did not develop MTJs. Muscle-FRT constructs in addition to muscle and tendon also demonstrated well developed cartilage, possessing high potential for development into bone. The muscle-FRT construct model could be used for studies of developmental mechanisms involved in the establishment of interfaces among all four muscularskeletal tissues: muscle, tendon and cartilage/bone.