Method for gene transfer
View Patent ↗Disclosed is a method for introducing a gene into a target cell using a retrovirus vector, which is simple and highly efficient. Specifically disclosed is a method for introducing a gene into a target cell using a retrovirus vector, which comprises the steps of (a) placing a liquor containing a retrovirus vector having a foreign gene carried thereon into a culture vessel on which a retrovirus-binding substance has been immobilized, and incubating the liquor at a temperature lower than 25° C. for 4 hours or longer, thereby producing a culture vessel having the retrovirus vector bound thereto, and (b) adding a target cell to the culture vessel that has been produced in step (a) and incubating the culture vessel. The gene introduction method is simple and highly efficient, and is useful particularly in the fields of medicine, cell technology, gene technology, embryologic technology and the like.
1. A method of transferring a foreign gene into a target cell using a retroviral vector, the method comprising the following steps (a) to (e):
(a) immobilizing a retrovirus-binding substance on an inner surface of a bag for cell culture;
(b) injecting a liquid containing a retroviral vector carrying a foreign gene into the bag obtained by the step (a), followed by incubation with shaking at a temperature of less than 25° C. for 8 hours to 48 hours to obtain a bag to which the retroviral vector is bound;
(c) adding a target cell into the bag obtained by the step (b), followed by incubation for 2 to 8 hours;
(d) turning the bag obtained by the step (c) upside down; and then
(e) incubating the bag obtained by the step (d) while upside down.
2. The method for transferring a gene according to claim 1 , further comprising a step of washing the bag obtained by the step (b) prior to the step (c).
3. The method according to claim 1 , wherein the retrovirus-binding substance is at least one selected from the group consisting of fibronectin, fibroblast growth factor, collagen type V, polylysine, DEAE-dextran, CH-296, H-271, H-296, and CH-271.
4. The method according to claim 1 , wherein the retrovirus-binding substance also has a cell-binding activity.
5. The method according to claim 1 , wherein the step (a) further comprises immobilizing a cell-binding substance on the inner surface of the bag.
6. The method according to claim 5 , wherein the cell-binding substance is at least one selected from the group consisting of a cell-adhesive protein, a hormone, a cytokine, an antibody, a sugar chain, and a carbohydrate.