Transfection of eukaryontic cells with linear polynucleotides by electroporation
AssigneesUniversity of Antwerp, Gerold Schuler, Isolde Strobel
InventorsGerold Schuler, Zwi N. Berneman, Viggo F. I. Van Tendeloo, Peter Ponsaerts, Isolde Strobel
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Abstract
The present invention provides an improved method for gene delivery in eukaryotic cells by electroporation, preferably in human hematopoietic cells, particular dendritic cells. The method of the invention is superior to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for gene delivery, including tumor antigen loading of dendritic cells.
Claims17
A method for transfection of hematopoietic cells or stem cells with one or more linear polynucleotides, which method comprises electroporation of a suspension containing the hematopoietic cells or stem cells and the linear polynucleotides to be transfected at a capacitance of 100 to below 300 μF.
A method for transfection of hematopoietic cells or stem cells with one or more linear polynucleotides, which method comprises electroporation of a suspension containing the hematopoietic cells or stem cells and the linear polynucleotides to be transfected with a soft pulse at 300 to 600 V for 100 us to 1 ms.
A method for the transfection of Mo-DC with linear mRNA, which method comprises electroporation of a suspension containing the Mo-DC, and the linear mRNA to be transfected, wherein the transfection is performed at a voltage of 250-300 V, a capacitance below 300 μF and a cell concentration of 1×107 to 4×107 cells/ml.
A method for the transfection of human mature Mo-DC with linear mRNA, which method comprises electroporation of a suspension containing the human mature Mo-DC and the linear mRNA to be transfected at a capacitance below 300 μF, wherein the method may further comprise further maturation of the transfected Mo-DC by providing a maturation stimulus.
A method for the transfection of human immature Mo-DC with linear mRNA, which method comprises electroporation of a suspension containing the human immature Mo-DC and the linear mRNA to be transfected at a capacitance below 300 μF, wherein the method may further comprise maturation of the transfected Mo-DC by providing a maturation-inducing compound.
Dependencies are inferred from the claim text.