Systems and methods for closed loop, real-time modifications of patient cells

A method of introducing a modification to a target nucleated blood cell, the method comprising:

• parenterally connecting a subject in need of such modification to a system via the inlet and outlet conduits, the system comprising:
• an inlet conduit adapted for parenteral communication with the subject and adapted for receiving blood from the subject,
• a cell separation module in fluid communication with the inlet conduit, the cell separation module comprising a cell separator configured to produce a fraction enriched in a target nucleated blood cell type using the blood from the subject, and the cell separation module,
• a cell customization module in fluid communication with the cell separation module so as to receive the nucleated blood cell fraction enriched in the target cell type from the cell separation module, the cell customization module configured to present one or more modifying agent to the target nucleated blood cells, wherein the cell customization module comprises a conduit that permits the target nucleated blood cells exposed to the one or more modifying agent to pass out of the cell customization module, and
• an outlet conduit in fluid communication with the cell separation module or the conduit of the cell customization module and adapted for parenteral communication with the subject, and the inlet conduit, the cell separation module, the cell customization module, and the outlet conduit of the system are connected in a fluid-sealed closed-loop;
• permitting the blood of the subject to flow into the cell separation module and producing a fraction enriched in the target nucleated blood cell type;
• permitting a blood cell fraction enriched in the target nucleated blood cell to flow from the cell separation module to the cell customization module and contacting the blood cell fraction enriched in the target nucleated blood cell type with one or more modifying agents;
• and
• permitting the target nucleated blood cells exposed to the one or more modifying agent to parenterally flow to the subject via the outlet conduit,
• wherein the target nucleated blood cell type is not cultured or otherwise expanded in population prior to the target nucleated blood cells exposed to the one or more modifying agent to parenterally flowing to the subject via the outlet conduit.