Method for gene transfer
Disclosed is a simple and highly efficient method for introducing a gene into a target cell using a retrovirus vector. The method comprises the steps of (a) placing a liquor containing a retrovirus vector having a foreign gene carried thereon into a bag for cell culture on which a retrovirus-binding substance has been immobilized, and incubating the liquor at a temperature lower than 25° C. for 8-48 hours, thereby producing a culture bag having the retrovirus vector bound thereto, (b) adding a target cell to the culture bag that has been produced in step (a) and incubating the culture bag for 8 hours or less, and (c) flipping the culture bag upside down and incubating the culture bag. The gene introduction method is useful particularly in medicine, cell technology, gene technology, and embryologic technology.
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) placing a liquid containing a retroviral vector carrying a foreign gene in the bag obtained by step (a), followed by incubation with shaking at a temperature of less than 25° C. for 8 to 48 hours to obtain the 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 1 to 4 hours;
(d) turning the bag obtained by step (c) upside down; and then
(e) incubating the bag obtained by step (d) while upside down.
2. The method according to claim 1 , wherein the bag is incubated for 1 to 2 hours in step (c).
3. The method according to claim 1 , further comprising a step of washing the bag obtained by the step (b) prior to the step (c).
4. 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, and a fibronectin fragment containing heparin-II domain.
5. The method according to claim 1 , wherein the retrovirus-binding substance also has a cell-binding activity.
6. The method according to claim 1 , wherein step (a) further comprises immobilizing a cell-binding substance on the inner surface of the bag.
7. The method according to claim 6 , 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.