Primary Cell

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Donald V. Lightner - One of the best experts on this subject based on the ideXlab platform.

  • Development of an in vitro Primary Cell culture system from the penaeid shrimp, Penaeus stylirostris and Penaeus vannamei
    Aquaculture, 1992
    Co-Authors: Renéa A. Luedeman, Donald V. Lightner
    Abstract:

    Abstract Studies were carried out to improve the methods for producing Primary Cell cultures of penaeid shrimp and to make these procedures routine and practical. Supplements tested included: lobster hemolymph, shrimp muscle extract, fetal bovine serum and hybridoma quality fetal bovine serum. Grace's Insect Medium supplemented with hybridoma quality fetal bovine serum was found to provide the best results. The optimum conditions for shrimp Cells in Grace's Insect Medium were determined to be an osmolality of approximately 700–750 mmol/kg, a temperature range of 25–28 °C and incubation in a normal atmosphere. Using the methods developed, monolayers of Primary cultures of ovarian epithelioid Cells from Penaeus stylirostris and Penaeus vannamei were routinely obtained with 80% confluence within a 2-day period.

Renéa A. Luedeman - One of the best experts on this subject based on the ideXlab platform.

  • Development of an in vitro Primary Cell culture system from the penaeid shrimp, Penaeus stylirostris and Penaeus vannamei
    Aquaculture, 1992
    Co-Authors: Renéa A. Luedeman, Donald V. Lightner
    Abstract:

    Abstract Studies were carried out to improve the methods for producing Primary Cell cultures of penaeid shrimp and to make these procedures routine and practical. Supplements tested included: lobster hemolymph, shrimp muscle extract, fetal bovine serum and hybridoma quality fetal bovine serum. Grace's Insect Medium supplemented with hybridoma quality fetal bovine serum was found to provide the best results. The optimum conditions for shrimp Cells in Grace's Insect Medium were determined to be an osmolality of approximately 700–750 mmol/kg, a temperature range of 25–28 °C and incubation in a normal atmosphere. Using the methods developed, monolayers of Primary cultures of ovarian epithelioid Cells from Penaeus stylirostris and Penaeus vannamei were routinely obtained with 80% confluence within a 2-day period.

Joseph R. Schulz - One of the best experts on this subject based on the ideXlab platform.

  • Primary Cell culture of adult zebrafish spinal neurons for electrophysiological studies.
    Journal of Neuroscience Methods, 2019
    Co-Authors: Max E. Meade, Jessica E. Roginsky, Joseph R. Schulz
    Abstract:

    Abstract Background Zebrafish (Danio rerio) are growing in popularity as a vertebrate model organism for the study of spinal neurocircuitry and locomotion. While many studies have used the zebrafish model system for electrophysiological analyses in embryonic and larval stages, there is a growing interest in studying spinal circuits and neurons from adult fish. New Method To expand upon the existing toolset available to the zebrafish research community, we have developed the first Primary Cell culture system of adult zebrafish spinal neurons. The intact spinal cord is dissected, and neurons are isolated through enzymatic digestion and mechanical dissociation. Identifiable neurons are viable for electrophysiological analyses after two days in culture. Results Spinal neurons in culture were confirmed by immunofluorescence labeling and found to exhibit distinct morphologies from other Cell types, allowing neurons to be identified based on morphology alone. Neurons were suitable for calcium imaging and whole Cell patch clamp recordings, which revealed excitable Cells with voltage-gated whole Cell currents, including tetrodotoxin-sensitive sodium currents. Comparison with Existing Methods This Primary Cell culture system is the only methodology available to isolate neurons from the adult zebrafish spinal cord. Other methods rely on keeping the spinal cord intact or the utilization of embryonic or larval stage fish. This method provides a robust platform for use in neurophysiological and pharmacological studies. Conclusions The novel Primary Cell culture system described here provides the first in vitro methodology available to isolate and culture neurons from the adult zebrafish spinal cord for use in electrophysiological analyses.

Olivier Lerouxel - One of the best experts on this subject based on the ideXlab platform.

  • Golgi-Mediated Synthesis and Secretion of Matrix Polysaccharides of the Primary Cell Wall of Higher Plants
    Frontiers in Plant Science, 2012
    Co-Authors: Azeddine Driouich, Marie-laure Follet-gueye, Sophie Bernard, Sumaira Kousar, Laurence Chevalier, Maïté Vicré-gibouin, Olivier Lerouxel
    Abstract:

    The Golgi apparatus of eukaryotic Cells is known for its central role in the processing, sorting , and transport of proteins to intra-and extra-Cellular compartments. In plants, it has the additional task of assembling and exporting the non-Cellulosic polysaccharides of the Cell wall matrix including pectin and hemiCelluloses, which are important for plant development and protection. In this review, we focus on the biosynthesis of complex polysaccharides of the Primary Cell wall of eudicotyledonous plants. We present and discuss the compartmen-tal organization of the Golgi stacks with regards to complex polysaccharide assembly and secretion using immuno-electron microscopy and specific antibodies recognizing various sugar epitopes. We also discuss the significance of the recently identified Golgi-localized glycosyltransferases responsible for the biosynthesis of xyloglucan (XyG) and pectin.

  • disrupting two arabidopsis thaliana xylosyltransferase genes results in plants deficient in xyloglucan a major Primary Cell wall component
    The Plant Cell, 2008
    Co-Authors: David Cavalier, Olivier Lerouxel, Lutz Neumetzler, Kazuchika Yamauchi, Antje Reinecke, Glenn Freshour, Olga A Zabotina, Michael G Hahn, Ingo Burgert, Markus Pauly
    Abstract:

    Xyloglucans are the main hemiCellulosic polysaccharides found in the Primary Cell walls of dicots and nongraminaceous monocots, where they are thought to interact with Cellulose to form a three-dimensional network that functions as the principal load-bearing structure of the Primary Cell wall. To determine whether two Arabidopsis thaliana genes that encode xylosyltransferases, XXT1 and XXT2, are involved in xyloglucan biosynthesis in vivo and to determine how the plant Cell wall is affected by the lack of expression of XXT1, XXT2, or both, we isolated and characterized xxt1 and xxt2 single and xxt1 xxt2 double T-DNA insertion mutants. Although the xxt1 and xxt2 mutants did not have a gross morphological phenotype, they did have a slight decrease in xyloglucan content and showed slightly altered distribution patterns for xyloglucan epitopes. More interestingly, the xxt1 xxt2 double mutant had aberrant root hairs and lacked detectable xyloglucan. The reduction of xyloglucan in the xxt2 mutant and the lack of detectable xyloglucan in the xxt1 xxt2 double mutant resulted in significant changes in the mechanical properties of these plants. We conclude that XXT1 and XXT2 encode xylosyltransferases that are required for xyloglucan biosynthesis. Moreover, the lack of detectable xyloglucan in the xxt1 xxt2 double mutant challenges conventional models of the plant Primary Cell wall.

Marie-christine Ralet - One of the best experts on this subject based on the ideXlab platform.

  • Competitive binding of pectin and xyloglucan with Primary Cell wall Cellulose
    Carbohydrate Polymers, 2008
    Co-Authors: Agata Zykwinska, Jean-françois Thibault, Marie-christine Ralet
    Abstract:

    Assemblies of pectin, xyloglucan and Cellulose were studied in vitro using two ternary systems. In the first one, xyloglucan concentration varied, while pectin amount was kept constant. In the second one, pectin concentration varied, whereas xyloglucan amount was fixed. The use of ternary systems allowed to put forward the hypothesis that pectin/Cellulose and xyloglucan/Cellulose associations may exist together or separately, depending on the proportion of non-Cellulosic polysaccharides in Cell walls. It can be hypothesized that pectin plays a double role within Primary Cell walls: (i) pectin loosely bound to Cellulose, in xyloglucan-rich Cell walls, (ii) pectin associated with Cellulose, in xyloglucan-poor Cell walls.