title: In Vitro Generation of Murine Dendritic Cells for Cancer Immunotherapy: An Optimized Protocol.
abstract: Dendritic cell (DC) immunotherapy induces tumor-reactive T-cells. We optimized the maturation of murine DC against ovarian cancer.
  
  
  
  Immature DC were generated from bone-marrow progenitor cells and loaded with hypericin-photodynamic-treated (Hyp-PDT) tumor cells (primary maturation). Lipopolysacharide (LPS 1 μg/ml) was used as a secondary maturation stimulus. After 24 h, maturation was assessed using flow cytometry. For in vivo experiments, C57BL/6 mice were vaccinated subcutaneously with matured, loaded mature DC. Immune response was evaluated through immunohistochemistry and cytometric bead array.
  
  
  
  Based on the existing protocols for DC generation, therapeutic vaccination of tumor-bearing mice with mature ID8-fLuc Hyp-PDT DC induces an immunogenic response, but provides no survival benefit. As loading with Hyp-PDT lysate induces partial primary maturation, we reduced the dose of LPS to 0.125 μg/ml and the duration of maturation to 6 hours, improving viability of mature DC.
  
  
  
  We optimized Hyp-PDT-based ID8-fLuc DC vaccine by reducing the amount of LPS and the duration of maturation.
authors: Department of Gynaecology and Obstetrics, Leuven Cancer Institute, University Hospitals Leuven, Leuven, Belgium thais.baert@gmail.com.|ImmunOvar Research Group, Laboratory for Tumor Immunology and Immunotherapy, Department of Oncology, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Thaïs
authors: T
authors: Baert
authors: Laboratory for Cell Death Research and Therapy, Department of Cellular and Molecular Medicine, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Abhishek D
authors: AD
authors: Garg
authors: ImmunOvar Research Group, Laboratory for Tumor Immunology and Immunotherapy, Department of Oncology, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Eva
authors: E
authors: Vindevogel
authors: Department of Gynaecology and Obstetrics, Leuven Cancer Institute, University Hospitals Leuven, Leuven, Belgium.|ImmunOvar Research Group, Laboratory for Tumor Immunology and Immunotherapy, Department of Oncology, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Anaïs
authors: A
authors: VAN Hoylandt
authors: Department of Development and Regeneration, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Godelieve
authors: G
authors: Verbist
authors: Laboratory for Cell Death Research and Therapy, Department of Cellular and Molecular Medicine, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Patrizia
authors: P
authors: Agostinis
authors: Department of Gynaecology and Obstetrics, Leuven Cancer Institute, University Hospitals Leuven, Leuven, Belgium.|ImmunOvar Research Group, Laboratory for Tumor Immunology and Immunotherapy, Department of Oncology, Organ Systems, KU Leuven, Leuven, Belgium.
authors: Ignace
authors: I
authors: Vergote
authors: Department of Gynaecology and Obstetrics, Leuven Cancer Institute, University Hospitals Leuven, Leuven, Belgium.|ImmunOvar Research Group, Laboratory for Tumor Immunology and Immunotherapy, Department of Oncology, Organ Systems, KU Leuven, Leuven, Belgium.
authors: A N
authors: AN
authors: Coosemans
mesh_terms: D000818:Animals; D003713:Dendritic Cells / Q000276:immunology*; D007167:Immunotherapy / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D009374:Neoplasms, Experimental / Q000628:therapy*
keywords: Dendritic cell; LPS; immunotherapy; maturation; mouse