Highly Efficient Transfection of Dendritic Cells Derived from Esophageal Squamous Cell Carcinoma Patient: Optimization with Green Fluorescent Protein and Validation with Tumor RNA as a Tool for Immuno-genetherapy

Authors

  • Ameneh Sazgarnia Department of Medical Physics, Avicenna Research Institute, MUMS, P.O. Box 9196773117, Mashhad, I.R. Iran
  • Mahmoud Mahmoudi Department of Molecular Immunology, Immunology Research Center, MUMS, P.O. Box 9196773117, Mashhad, I.R. Iran
  • Marta Ghahraman Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences (MUMS), P.O. Box 9196773117, Mashhad, I.R. Iran
  • Mehran Gholamin Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences (MUMS), P.O. Box 9196773117, Mashhad, I.R. Iran
  • Moein Farshchian Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences (MUMS), P.O. Box 9196773117, Mashhad, I.R. Iran
  • Mohammad Reza Abbaszadegan Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences (MUMS), P.O. Box 9196773117, Mashhad, I.R. Iran
  • Mohammad Taghi Rajabi-Mashhadi Department of Surgery, Omid Hospital, MUMS, P.O. Box 13131-91389, Mashhad, Iran
  • Mojtaba Sankian Department of Immunobiochemistry, Immunology Research Center, MUMS, P.O. Box 9196773117, Mashhad, I.R. Iran
  • Omeed Moaven Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences (MUMS), P.O. Box 9196773117, Mashhad, I.R. Iran
Abstract:

This study was conducted to optimize a highly efficient mRNA transfection into dendritic cells (DC) derived from esophageal squamous cell carcinoma (ESCC) patients. Applying an electroporation technique, in vitro synthesized Green Fluorescent Protein (GFP) mRNA was transfected as an indicator into the DCs derived from a healthy donor. Flow cytometry revealed 84.9% transfection efficiency for DCs transfected with GFP mRNA. Optimized condition (500V/300 ms) yielded 79.8% efficiency in transfecting GFP mRNA into DCs from ESCC patient. Applying this efficient method, tumoral mRNA was transfected into DCs. T cells were then primed with tumor RNA/DCs and cytotoxicity assay revealed significantly higher lyses of tumor/DC vs. Mock DC; approving the optimized method for further establishment of the preclinical phase of DC-based immunotherapy for ESCC.

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volume 8  issue 2

pages  121- 126

publication date 2010-04-01

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