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Novel non-viral method for transfection of primary leukemia cells and cell lines
BACKGROUND: Tumor cells such as leukemia and lymphoma cells are possible targets for gene therapy. However, previously leukemia and lymphoma cells have been demonstrated to be resistant to most of non-viral gene transfer methods. METHODS: The aim of this study was to analyze various methods for tran...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2004
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC331421/ https://www.ncbi.nlm.nih.gov/pubmed/14715084 http://dx.doi.org/10.1186/1479-0556-2-1 |
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author | Schakowski, Frank Buttgereit, Peter Mazur, Martin Märten, Angela Schöttker, Björn Gorschlüter, Marcus Schmidt-Wolf, Ingo GH |
author_facet | Schakowski, Frank Buttgereit, Peter Mazur, Martin Märten, Angela Schöttker, Björn Gorschlüter, Marcus Schmidt-Wolf, Ingo GH |
author_sort | Schakowski, Frank |
collection | PubMed |
description | BACKGROUND: Tumor cells such as leukemia and lymphoma cells are possible targets for gene therapy. However, previously leukemia and lymphoma cells have been demonstrated to be resistant to most of non-viral gene transfer methods. METHODS: The aim of this study was to analyze various methods for transfection of primary leukemia cells and leukemia cell lines and to improve the efficiency of gene delivery. Here, we evaluated a novel electroporation based technique called nucleofection. This novel technique uses a combination of special electrical parameters and specific solutions to deliver the DNA directly to the cell nucleus under mild conditions. RESULTS: Using this technique for gene transfer up to 75% of primary cells derived from three acute myeloid leukemia (AML) patients and K562 cells were transfected with the green flourescent protein (GFP) reporter gene with low cytotoxicity. In addition, 49(+/- 9.7%) of HL60 leukemia cells showed expression of GFP. CONCLUSION: The non-viral transfection method described here may have an impact on the use of primary leukemia cells and leukemia cell lines in cancer gene therapy. |
format | Text |
id | pubmed-331421 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2004 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-3314212004-02-07 Novel non-viral method for transfection of primary leukemia cells and cell lines Schakowski, Frank Buttgereit, Peter Mazur, Martin Märten, Angela Schöttker, Björn Gorschlüter, Marcus Schmidt-Wolf, Ingo GH Genet Vaccines Ther Research BACKGROUND: Tumor cells such as leukemia and lymphoma cells are possible targets for gene therapy. However, previously leukemia and lymphoma cells have been demonstrated to be resistant to most of non-viral gene transfer methods. METHODS: The aim of this study was to analyze various methods for transfection of primary leukemia cells and leukemia cell lines and to improve the efficiency of gene delivery. Here, we evaluated a novel electroporation based technique called nucleofection. This novel technique uses a combination of special electrical parameters and specific solutions to deliver the DNA directly to the cell nucleus under mild conditions. RESULTS: Using this technique for gene transfer up to 75% of primary cells derived from three acute myeloid leukemia (AML) patients and K562 cells were transfected with the green flourescent protein (GFP) reporter gene with low cytotoxicity. In addition, 49(+/- 9.7%) of HL60 leukemia cells showed expression of GFP. CONCLUSION: The non-viral transfection method described here may have an impact on the use of primary leukemia cells and leukemia cell lines in cancer gene therapy. BioMed Central 2004-01-12 /pmc/articles/PMC331421/ /pubmed/14715084 http://dx.doi.org/10.1186/1479-0556-2-1 Text en Copyright © 2004 Schakowski et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. |
spellingShingle | Research Schakowski, Frank Buttgereit, Peter Mazur, Martin Märten, Angela Schöttker, Björn Gorschlüter, Marcus Schmidt-Wolf, Ingo GH Novel non-viral method for transfection of primary leukemia cells and cell lines |
title | Novel non-viral method for transfection of primary leukemia cells and cell lines |
title_full | Novel non-viral method for transfection of primary leukemia cells and cell lines |
title_fullStr | Novel non-viral method for transfection of primary leukemia cells and cell lines |
title_full_unstemmed | Novel non-viral method for transfection of primary leukemia cells and cell lines |
title_short | Novel non-viral method for transfection of primary leukemia cells and cell lines |
title_sort | novel non-viral method for transfection of primary leukemia cells and cell lines |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC331421/ https://www.ncbi.nlm.nih.gov/pubmed/14715084 http://dx.doi.org/10.1186/1479-0556-2-1 |
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