T Cell Expansion in the Presence of RetroNectin® Reagent

29 Sept 2014

RetroNectin reagent is used widely to enhance retroviral/lentiviral gene transfer into hematopoietic stem cells, lymphocytes, and other blood cells. This application note demonstrates an additional use of RetroNectin reagent for T lymphocyte expansion from peripheral blood mononuclear cells.

RetroNectin® Reagent

Takara Bio

Research and Clinical Grade RetroNectin Reagent for Retroviral or Lentiviral Transduction. RetroNectin Reagent for retroviral or lentiviral transduction significantly increases transduction efficiency and results in enhanced gene transfer, especially in suspension and hematopoietic stem cells. RetroNectin Reagent is a chimeric peptide composed of E. coli-derived recombinant human fibronectin fragment that contains three functional domains: a central cell-binding domain (type III repeat, 8-10), a heparin-binding domain II (type III repeat, 12-14), and a CS1 site within the alternatively-spliced IIICS region. RetroNectin is a 574 amino acid protein with a molecular weight of 63 kDa. Fibronectins (FNS) are multi-functional cell adhesive glycoproteins present in the extracellular cell matrix and plasma and also play critical roles in wound healing and embryogenesis.When coated on surfaces of cell culture dishes, flasks or bags, RetroNectin significantly increases retroviral or lentiviral transduction into mammalian cells. This enhanced gene transfer is thought to occur via the co-localization of retroviral particles and target cells on the RetroNectin molecules. In essence, viral particles bind RetroNectin via interaction with its heparin-binding domain II, while target cells bind RetroNectin primarily through interaction of the cell surface integrin receptor VLA-4 with the fibronectin CS1 site. Additionally, target cells may also bind the RetroNectin recombinant human fibronectin fragment via the interaction of fibronectin tripeptide Arg-Gly-Asp (RGD), located within the molecule's central cell-binding domain, with corresponding cell surface integrin receptor VLA-5.Features: Enhanced gene transfer efficiency Higher retroviral or lentiviral transduction levels in suspended and adherent mammalian cells Improved success in hard-to-infect cell types and hematopoietic stem cells Nontoxic: does not require cytotoxic polybrene Superior transduction efficiency compared with fibronectin, polybrene or protamine Higher reagent efficiency enables use of smaller viral solution volumes Clinical grade RetroNectin is available Applications: General retroviral or lentiviral transduction experiments Cell migration assays Gene therapy studies: RetroNectin has already been used in over 40 clinical trials, mostly using hematopoietic stem cells Cell therapy studies related to stem cell research and related technology

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Cell / Tissue CultureCell culture or tissue culture is used to study the biology of cells or tissues and to isolate cellular products in an environment which can be manipulated and well defined. Accurately control your culture environment with bioreactors or culture incubators, bind your cells to a surface or together with an extracellular matrix. Distinguish cell types with differential media or proliferate cells with certain characteristics using selective media. Enrich your media with supplements such as growth factors, sera and vitamins. Find the best cell and tissue culture products, kits and equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ImmunologyImmunological techniques measure and characterize immune responses. Immunology kits and analysis systems often use techniques such as ELISA, radioimmunoassay (RIA) and immunodiffusion assays, Immunohistochemistry, and flow cytometry. Immunologists use equipment such as flow Cytometers, plate readers, plate washers and fluorescent microscopes.
T Cell Expansion in the Presence of RetroNectin® Reagent