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

Author(s):

Mohammad Reza Abbaszadegan , Mehran Gholamin , Omeed Moaven , Moein Farshchian , Mohammad Taghi Rajabi , Mahmoud Mahmoudi , Mojtaba Sankian , Ameneh Sazegarnia , Marta Ghahraman , Abbas Abdollahi

Message:
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.Keywords: Immuno-gene therapy; Esophageal squamous cell carcinoma (ESCC); Electroporation; mRNA transfection; Dendritic cell; GFP
Language:
English
Published:
Iranian Journal of Biotechnology, Volume:8 Issue: 2, Spring 2010
Pages:
121 to 126
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