A closed system for labelling and selecting live cells

A method for efficient viral-mediated gene transfer in mammalian cells comprising:

• a. Providing a first input bag containing a mammalian cell population and a second input bag containing a transduction buffer comprising a concentrated viral vector that is packaged with genetic material foreign to the mammalian cell population;
• b. Adding the first input bag containing the mammalian cell population and the transduction buffer comprising a concentrated viral vector that is packaged with genetic material foreign to the mammalian cell population to a chamber embedded in a centrifuge rotor while the rotor is in motion and a counterflow in the chamber produces an opposing force within the chamber;
• c. Incubating the mammalian cell population with the concentrated viral vector packaged with genetic material foreign to the mammalian cell population by circulating the transduction buffer comprising the concentrated viral vector that is packaged with the genetic material of interest around the cells, wherein the incubating is effective to transfer genetic material from the viral vector to a subpopulation of the mammalian cell population to form a transfected subpopulation of mammalian cells;
• d. selectively labeling the transfected subpopulation of mammalian cells by
• (i) incubating the mammalian cell population with a capture particle comprising an agent that recognizes and binds specifically a cell antigen expressed selectively by the transfected subpopulation within the heterogeneous cell population;
• (ii) binding the capture particle comprising the agent to the targeted population of cells, to form a labeled transfected subpopulation of cells;
• e. passing a wash buffer through the chamber embedded in the centrifuge rotor while the rotor is in motion and the counterflow produces an opposing force within the chamber, wherein the wash buffer removes unbound cells and unbound capture particles from the chamber;
• f. collecting in an output bag the transfected subpopulation of cells bound to the capture particle comprising the agent that recognizes the specific cell surface marker so that the cells bound to the agent that recognizes the specific cell surface marker are enriched relative to the heterogeneous cell suspension;
• and
• g. dissociating the cells in (f) from the agent that recognizes the specific cell surface marker,
• wherein the method is effective to:
• (i) reduce the risk of contamination of the collected cells;
• (ii) reduce damage to the collected cells;
• (iii) maintain viability of the collected cells;
• or
• (iv) a combination thereof.