Retroviral transduction using poloxamers
The present invention relates to a method for transducing a target cell, the method comprising the step of contacting a target cell with a retroviral vector and a poloxamer having a molecular weight of 12.8 kDa to about 15 kDa. Further, the invention relates to the use of a poloxamer as defined herein, optionally in combination with a polycationic substance as defined herein, for transducing a target cell with a retroviral vector and a kit comprising a retroviral vector, a poloxamer as defined herein and, optionally, instructions for use.
1. A method for transducing a target cell, the method comprising the step of contacting a target cell ex vivo or in vitro with a retroviral vector and a poloxamer having a molecular weight of 12.8 kDa to about 15 kDa, wherein the poloxamer is in a fluid state during transduction of the target cell.
2. The method of claim 1 , wherein the target cell is a cell selected from the group consisting of a lymphocyte, a tumor cell, a lymphoid lineage cell, a neuronal cell, an epithelial cell, an endothelial cell, a primary cell, and a stem cell.
3. The method of claim 2 , wherein the lymphocyte is a primary lymphocyte and/or wherein the tumor cell is a hematopoietic tumor cell, a neuronal tumor cell or an epithelial tumor cell.
4. The method of any one of claims 1 to 3 , wherein the retroviral vector is a lentiviral vector.
5. The method of claim 4 , wherein the lentiviral vector is pseudotyped with at least one vesicular stomatitis virus glycoprotein (VSV-G) and/or with an antibody fragment fused to VSV-G.
6. The method of claim 1 , wherein the poloxamer has the formula HO—[CH2CH2O]x-[CH2C2H4O]z-[CH2CH2O]y, wherein x+y=265.45 and z=50.34 on average; or wherein the poloxamer has the formula HO—[CH2CH2O]x-[CH2C2H4O]z-[CH2CH20]y, wherein x+y=236.36 and z=44.83 on average.
7. The method of claim 1 , wherein said target cell is further brought into contact with one or more polycationic substances selected from the group consisting of polycationic polymers or polycationic peptides.
8. The method of claim 7 , wherein said polycationic polymers are selected from the group consisting of poly(ethylene glycol)-poly(L-lysine) block copolymer (PEG-PLL) and 1,5-dimethyl-1,5-diaza-undeca-methyl-polymethobromide (Polybrene); and/or said polycationic peptides are selected from the group consisting of protamine sulphate and poly-1-lysin (PLL) having a mean molecular weight from 1 to 300 kDa.
9. The method of claim 8 , wherein the polycationic substances are 1,5-dimethyl-1,5-diaza-undeca-methyl-polymethobromide and/or protamine sulphate.
10. The method of 1 , wherein said poloxamer is provided at a concentration of about 50 to 5000 μg/ml.
11. The method of claim 10 , wherein said poloxamer is provided at a concentration of about 500 to 1000 μg/ml.
12. The method of claim 1 , comprising the further step of spinoculating said retroviral vector with said target cell prior to, concomitant with or after contacting said target cell with said poloxamer.
13. The method of claim 1 , wherein the poloxamer is in a fluid state during the contacting step.