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The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

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प्राथमिक ट्यूमर से प्रतिरक्षा कोशिकाओं का अलगाव

1, 1, 2, 1

1Tumor Immunity and Tolerance Section, Laboratory of Molecular Immunoregulation, Cancer and Inflammation Program, National Cancer Institute - Frederick, 2KEWB Productions

 

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Cite this Article: प्राथमिक ट्यूमर से प्रतिरक्षा कोशिकाओं का अलगाव

Watkins, S. K., Zhu, Z., Watkins, K. E., Hurwitz, A. A. Isolation of Immune Cells from Primary Tumors. J. Vis. Exp. (64), e3952, doi:10.3791/3952 (2012).

Disclosures: प्राथमिक ट्यूमर से प्रतिरक्षा कोशिकाओं का अलगाव

Materials: प्राथमिक ट्यूमर से प्रतिरक्षा कोशिकाओं का अलगाव

Name Company Catalog Number Comments
Collagenase I GIBCO, by Life Technologies 17100-017 Use when selecting for T cells
Collagenase IV GIBCO, by Life Technologies 17104-019 Use for myeloid selection
DNase Calbiochem 260913
RPMI GIBCO, by Life Technologies 21870
Dulbecco’s PBS Lonza Inc. 17-5158
Fetal Bovine Serum Lonza Inc. 14-501F
MACs Buffer 2% FBS for T cells
10% FBS for myeloid cells
Thy1.1 PE eBioscience 551401
Anti-PE microbeads Miltenyi Biotec 120-000-294
Anti-Pan DC microbeads Miltenyi Biotec 120-003-183
Anti-CD11b microbeads Miltenyi Biotec 120-000-300
LC column Miltenyi Biotec 130-042-202
MS column Miltenyi Biotec 130-042-201

References: प्राथमिक ट्यूमर से प्रतिरक्षा कोशिकाओं का अलगाव

  1. Shurin, M.R., et al. Intratumoral cytokines/chemokines/growth factors and tumor infiltrating dendritic cells: friends or enemies? Cancer Metastasis Rev. 25, 333-356 (2006).
  2. Lewis, C.E. & Pollard, J.W. Distinct role of macrophages in different tumor microenvironments. Cancer Res. 66, 605-612 (2006).
  3. Anderson, M.J., Shafer-Weaver, K., Greenberg, N.M., & Hurwitz, A.A. Tolerization of tumor-specific T cells despite efficient initial priming in a primary murine model of prostate cancer. J. Immunol. 178, 1268-1276 (2007).
  4. Ganss, R. & Hanahan, D. Tumor microenvironment can restrict the effectiveness of activated antitumor lymphocytes. Cancer Res. 58, 4673-4681 (1998).
  5. Shafer-Weaver, K.A., et al. Immunity to murine prostatic tumors: continuous provision of T-cell help prevents CD8 T-cell tolerance and activates tumor-infiltrating dendritic cells. Cancer Research. 69, 6256-6264 (2009).
  6. Watkins, S.K., Zhu, Z., Riboldi, E., Shafer-Weaver, K.A., Stagliano, K.E.R., Sklavos, M.M., Ambs, S., Yagita, H., & Hurwitz, A.A. Foxo3a programs tumor associated dendritic cells to become tolerogenic in Human and Murine Prostate Cancer. Journal of Clinical Investigation., 121 (2011).
  7. Shafer-Weaver, K.A. & Hurwitz, A.A. Tumor-Specific CD8+ T Cells Infiltrating Prostatic Tumors are Induced to Become Suppressor Cells., (2009).
  8. Singh, V., Ji, Q., Feigenbaum, L., Leighty, R.M., & Hurwitz, A.A. Melanoma progression despite infiltration by in vivo-primed TRP-2-specific T cells. J. Immunother. 32, 129-139 (2009).
  9. Matheu, M.P. & Cahalan, M.D. Isolation of CD4+ T cells from Mouse Lymph Nodes Using Miltenyi MACS Purification. J. Vis. Exp. (9), e409, DOI: 10.3791/409 (2007).
  10. Hurwitz, A.A., Foster, B.A., Allison, J.P., Greenberg, N.M., & Kwon, E.D. The TRAMP mouse as a model for prostate cancer. Curr.Protoc.Immunol. Chapter 20, Unit (2001).

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