We are analyzing https://link.springer.com/protocol/10.1007/978-1-59745-409-4_7.

Title:
Retroviral Transduction of Murine Primary T Lymphocytes | SpringerLink
Description:
In comparison to human T cells, efficient retroviral gene transfer and subsequent expansion of murine primary T cells is more difficult to achieve. Herein, we describe an optimized gene transfer protocol utilizing an ecotropic viral vector to transduce primary murine...
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Keywords {🔍}

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Topics {✒️}

basic amino-acid transporter primary t-lym-phocytes high-efficiency gene transfer systemic b-cell tumors tumor-infiltrating lymphocytes transduced retrovirus-mediated gene transfer murine primary t-lymphocytes magnetic beads coated hematopoietic stem cells gene therapy protocols high gene transfer privacy choices/manage cookies genetically modified cells protocol genetic modification agonistic anti-cd3 author information authors genetically targeted human optimized retroviral transduction adenosine deaminase deficiency human adenosine deaminase cellular gene therapy experimental therapeutics center journal finder publish conditions privacy policy ecotropic murine retroviruses european economic area ecotropic viral vector cell spin-oculation cytotoxic antiviral immunity de vries cr rogers-freezer lj helper-free retroviruses cell-surface receptor adenosine deaminase-deficient recombinant fibronectin fragments carl-neuberg-straβe 1 transduce primary murine national cancer institute commonwealth cancer foundation genetically engineered lymphocytes humana press accepting optional cookies hannover medical school main content log genetic modification cancer research check access current protocols protocol lee ethics access

Schema {🗺️}

ScholarlyArticle:
      headline:Retroviral Transduction of Murine Primary T Lymphocytes
      pageEnd:96
      pageStart:83
      image:https://media.springernature.com/w153/springer-static/cover/book/978-1-59745-409-4.jpg
      genre:
         Springer Protocols
      isPartOf:
         name:Genetic Modification of Hematopoietic Stem Cells
         isbn:
            978-1-59745-409-4
            978-1-58829-980-2
         type:Book
      publisher:
         name:Humana Press
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:James Lee
            affiliation:
                  name:Memorial Sloan-Kettering Cancer Center
                  address:
                     name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Michel Sadelain
            affiliation:
                  name:Memorial Sloan-Kettering Cancer Center
                  address:
                     name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Renier Brentjens
            affiliation:
                  name:Memorial Sloan-Kettering Cancer Center
                  address:
                     name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
                     type:PostalAddress
                  type:Organization
            type:Person
      keywords:Retroviral gene transfer, Murine primary T cells, Mouse T lymphocytes, Phoenix-eco packaging cells, Spinoculation, RetroNectin, Interleukin-2 (IL-2), Nylon wool column, CD3/ CD28-activating magnetic beads
      description:In comparison to human T cells, efficient retroviral gene transfer and subsequent expansion of murine primary T cells is more difficult to achieve. Herein, we describe an optimized gene transfer protocol utilizing an ecotropic viral vector to transduce primary murine T cells activated with magnetic beads coated with agonistic anti-CD3 and CD28 antibodies. Activated T cells are subsequently centrifuged (spinoculated) on RetroNectin-coated tissue culture plates in the context of retroviral supernatant. Variables found to be critical to high gene transfer and subsequent efficient T cell expansion included CD3/CD28 magnetic bead to cell ratio, time from T cell activation to initial spinoculation, frequency of T cell spin-oculation, interleukin-2 concentration in the medium, and the initial purity of the T cell preparation.
      datePublished:2009
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Book:
      name:Genetic Modification of Hematopoietic Stem Cells
      isbn:
         978-1-59745-409-4
         978-1-58829-980-2
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      name:Humana Press
      logo:
         url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
         type:ImageObject
      name:Memorial Sloan-Kettering Cancer Center
      address:
         name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
         type:PostalAddress
      name:Memorial Sloan-Kettering Cancer Center
      address:
         name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
         type:PostalAddress
      name:Memorial Sloan-Kettering Cancer Center
      address:
         name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
         type:PostalAddress
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      name:James Lee
      affiliation:
            name:Memorial Sloan-Kettering Cancer Center
            address:
               name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
               type:PostalAddress
            type:Organization
      name:Michel Sadelain
      affiliation:
            name:Memorial Sloan-Kettering Cancer Center
            address:
               name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
               type:PostalAddress
            type:Organization
      name:Renier Brentjens
      affiliation:
            name:Memorial Sloan-Kettering Cancer Center
            address:
               name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
               type:PostalAddress
            type:Organization
PostalAddress:
      name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
      name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
      name:Department of Medicine and Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA
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