Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells
| Autor: | Niek P. van Til, Marshall W. Huston, Gerard Wagemaker, J. Henk de Jong, Ian C.D. Johnston, Arzu Cengizeroglu, Yolanda Sanchez-Antequera, Zygmunt Pojda, Martina Anton, Christian Plank, Olga Mykhaylyk |
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| Přispěvatelé: | Hematology |
| Rok vydání: | 2011 |
| Předmět: |
Immunology
Cell Population Genetic Vectors 02 engineering and technology Cell Separation Gene delivery Biology Transfection Biochemistry Jurkat cells 03 medical and health sciences Transduction (genetics) Jurkat Cells Magnetics Mice medicine Animals Antigens Ly Humans education 030304 developmental biology 0303 health sciences education.field_of_study Mesenchymal stem cell Gene Transfer Techniques Hematopoietic stem cell Membrane Proteins Mesenchymal Stem Cells Cell Biology Hematology 021001 nanoscience & nanotechnology Hematopoietic Stem Cells Molecular biology medicine.anatomical_structure Nanoparticles 0210 nano-technology K562 Cells Interleukin Receptor Common gamma Subunit |
| Zdroj: | Blood, 117(16), E171-E181. American Society of Hematology Blood; Vol 117 |
| ISSN: | 1528-0020 0006-4971 |
| DOI: | 10.1182/blood-2010-08-302646 |
| Popis: | Research applications and cell therapies involving genetically modified cells require reliable, standardized, and cost-effective methods for cell manipulation. We report a novel nanomagnetic method for integrated cell separation and gene delivery. Gene vectors associated with magnetic nanoparticles are used to transfect/transduce target cells while being passaged and separated through a high gradient magnetic field cell separation column. The integrated method yields excellent target cell purity and recovery. Nonviral and lentiviral magselectofection is efficient and highly specific for the target cell population as demonstrated with a K562/Jurkat T-cell mixture. Both mouse and human enriched hematopoietic stem cell pools were effectively transduced by lentiviral magselectofection, which did not affect the hematopoietic progenitor cell number determined by in vitro colony assays. Highly effective reconstitution of T and B lymphocytes was achieved by magselectofected murine wild-type lineage-negative Sca-1+ cells transplanted into Il2rg−/− mice, stably expressing GFP in erythroid, myeloid, T-, and B-cell lineages. Furthermore, nonviral, lentiviral, and adenoviral magselectofection yielded high transfection/transduction efficiency in human umbilical cord mesenchymal stem cells and was fully compatible with their differentiation potential. Upscaling to a clinically approved automated cell separation device was feasible. Hence, once optimized, validated, and approved, the method may greatly facilitate the generation of genetically engineered cells for cell therapies. |
| Databáze: | OpenAIRE |
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