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      Estudio de cultivos celulares primarios derivados de Lucilia sericata (Diptera: Calliphoridae) Translated title: Study of primary cell cultures derived from Lucilia sericata (Diptera: Calliphoridae)

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          Abstract

          El propósito principal de la investigación aquí presentada fue obtener cultivos celulares primarios derivados de Lucilia sericata (Diptera: Calliphoridae). Esta mosca necrófaga es utilizada para determinar el intervalo post-mortem y en terapia larval. A partir de huevos embrionados, se realizaron explantes en diversos medios de cultivo (Grace, Schneider, MM/VP12, DMEM, Grace/L-15 y L-15), suplementados con 20% de suero fetal bovino. La esterilización del material biológico se efectuó mediante la aplicación de soluciones de formaldehido e hipoclorito de sodio. El crecimiento celular se inicio en los medios L-15, MM/VP12, Grace/L-15 y Schneider, en un tiempo promedio de 10 días después de efectuadas las siembras de tejidos embrionarios, mediante la proliferación de grupos de colonias dispersas en la superficies de los frascos de cultivo y a partir de las terminaciones de los fragmentos larvales. La evolución del crecimiento celular hasta la formación de la monocapa semiconfluente fue relativamente rápida, se alcanzo a las tres semanas post-explantes. La morfología de las células en los cultivos fue heterogénea, se destacaron formas epitelioides, similares a nerviosas, gigantes e irregulares. La comparación de las características de crecimiento de los cultivos celulares de L. sericata con los obtenidos de otras especies de dípteros mostro mayor favorabilidad en la evolución, en razón a que las células se adaptaron mejor a las condiciones fisico-quimicas de varios medios de cultivo. Este es el primer informe de cultivos celulares de una mosca de la familia Calliphoridae.

          Translated abstract

          The main purpose of this study was to obtain primary cell cultures derived from Lucilia sericata (Diptera: Calliphoridae). Necrophagous this fly is used for determination of post-mortem interval and larval therapy. Since explants embryonated eggs were performed in various culture media (Grace Schneider, MM/VP12, DMEM, Grace/L-15 and L-15), supplemented with 20% fetal serum. Sterilization of the biological material was carried out by immersing it in formaldehyde and sodium hypochlorite solutions. The cell growth was initiated in the L-15, MM/VP12, and Schneider Grace/L-15 in an average time of 10 days after completion of planting by the proliferation of groups of colonies scattered on the surface of the boxes crops and also from the endings of larval fragments. The evolution of cell growth to the formation of monolayer semi-confluent was relatively fast, reaching at 3 weeks post-explant. Cellular morphology in cultured cells was heterogeneous, especially epithelioid forms, similar to nerve, giant and irregular. Comparison of the growth characteristics of these cell cultures with those obtained from other species of flies was more favorable in the evolution of those obtained from L. sericata, on the grounds that the cells are better adapted to the physical-chemical conditions of several culture media. This is the first report of a cell culture-fly family Calliphoridae.

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          Effect of temperature on Lucilia sericata (Diptera: Calliphoridae) development with special reference to the isomegalen- and isomorphen-diagram.

          Developmental behavior of eggs, larva and pupa of the blowfly species Lucilia sericata (Meigen) were studied under 10 different temperature regimes. Data from these studies were used to construct the isomegalen-diagram. In this diagram, time from hatching to peakfeeding is plotted against temperature, each line representing identical larval length at various temperatures. If the temperature is roughly constant, as is the case with corpses found indoors, the age of the maggot can be read off instantly from its length, provided that the maggot has not entered the migratory phase. Where temperature is variable, an age range can be estimated between the points where the measured larval length cuts the graph at the maximum and minimum temperatures recorded. Equally, the isomorphen-diagram representing all morphological stages from oviposition to eclosion should be used, if maggots in the migratory phase or pupae or puparia are recovered from the scene. The isomegalen- and the isomorphen-diagrams could facilitate a quick and more precise estimate of the postmortem interval even for the inexperienced investigator. In addition, our results vary from those of other investigators, suggesting a different thermal behavior of the holarctic blowfly L. sericata in various zoogeographic regions.
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            Establishment of four strains of cells from insect tissues grown in vitro.

            T. Grace (1962)
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              Insect cell culture for industrial production of recombinant proteins.

              Insect cells used in conjunction with the baculovirus expression vector system (BEVS) are gaining ground rapidly as a platform for recombinant protein production. Insect cells present several comparative advantages to mammalian cells, such as ease of culture, higher tolerance to osmolality and by-product concentration and higher expression levels when infected with a recombinant baculovirus. Here we review some of the recent developments in protein expression by insect cells and their potential application in large-scale culture. Our current knowledge of insect cell metabolism is summarised and emphasis is placed on elements useful in the rational design of serum-free media. The culture of insect cells in the absence of serum is reaching maturity, and promising serum substitutes (hydrolysates, new growth and production-enhancing factors) are being evaluated. Proteolysis is a problem of the BEVS system due to its lytic nature, and can, therefore, be a critical issue in insect cell bioprocessing. Several cell- or baculovirus proteases are involved in degradation events during protein production by insect cells. Methods for proteolysis control, the optimal inhibitors and culture and storage conditions which affect proteolysis are discussed. Finally, engineering issues related to high-density culture (new bioreactor types, gas exchange, feeding strategies) are addressed in view of their relevance to large-scale culture.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                recis
                Revista Ciencias de la Salud
                Rev. Cienc. Salud
                Editorial Universidad del Rosario (Bogotá )
                1692-7273
                December 2009
                : 7
                : 3
                : 17-28
                Affiliations
                [1 ] Universidad del Rosario Colombia
                [2 ] Universidad de La Salle Colombia
                [3 ] Universidad del Rosario Colombia
                [4 ] Universidad del Rosario Colombia
                Article
                S1692-72732009000300003
                f9a9e081-9517-428e-9539-0ab8a205b10f

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Colombia

                Self URI (journal page): http://www.scielo.org.co/scielo.php?script=sci_serial&pid=1692-7273&lng=en
                Categories
                PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

                Public health
                Lucilia sericata,cellular cultures,cellular morphometry,cultivos celulares,morfometría celular

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