A method of collecting mononuclear cells includes separating plasma from cellular components of whole blood. The cellular components, which include mononuclear cells and red blood cells, are combined with plasma replacement fluid to form a first mixture. The mononuclear cells of the first mixture are separated from the red blood cells of the first mixture, which may be followed by extracorporeal photopheresis being performed on the mononuclear cells.

Blood in a separation chamber is separated into a red blood cell layer, a plasma constituent, and a mononuclear cell-containing layer. A portion of the plasma constituent exits the chamber via a plasma outlet, while a first portion of the red blood cell layer exits via a red blood cell outlet. A second portion of the red blood cell layer exits the chamber via the red blood cell outlet and is collected. At least a portion of the collected red blood cell layer may then be conveyed to the chamber via the red blood cell outlet to convey at least a portion of the mononuclear cell-containing layer out of the chamber via the plasma outlet for collection. A second portion of the plasma constituent may be conveyed out of the chamber via the plasma outlet to more fully collect the mononuclear cell-containing layer without the use of collected plasma.

The present invention relates to stem-cell derived hematopoietic cells, in particular, myeloid cells, preferably, macrophages, their generation and use. In particular, the invention relates to a method of producing hematopoietic, preferably, myeloid cells, comprising cultivating embryoid bodies, which are, e.g., derivable from pluripotent stem cells such as induced pluripotent stem cells (iPSC), in suspension culture, to produce myeloid cell forming complexes, which are further cultivated in suspension culture to produce myeloid cells such as macrophages. This allows for a scalable and continuous production, e.g., in industry-compatible stirred tank bioreactors. Macrophages, e.g., macrophages produced with this method that have unique characteristics, can be used in pharmaceutical compositions for treatment of patients, e.g., for treatment of infection such as bacterial infection. The invention further provides application systems suitable for spraying, comprising myeloid cells such as macrophages, which can have a reduced size, for use in treatment of patients for treatment of infection, e.g., bacterial infection or wound healing.

An extracorporeal photopheresis system includes a separator with a disposable fluid circuit including a treatment container, an irradiation device configured to treat the contents of the treatment container, and a controller configured to control the system to perform a procedure including drawing anticoagulated whole blood into the fluid circuit from a blood source and returning to the blood source a treated target cell component, a portion of a red blood cell component remaining in the fluid circuit, and/or a portion of a plasma component remaining in the fluid circuit. The controller is further configured to estimate an end-of-procedure fluid balance estimated based on manual or automatic inputs including a patient body weight associated with the blood source and a total blood volume of the blood source, indicate the fluid balance to an operator, and receive one or more changes that affect the fluid balance after indicating the fluid balance.

A method of collecting mononuclear cells, comprising separating whole blood into cellular components and platelets suspended in plasma, separating the platelets suspended in plasma into platelet concentrate and platelet-poor plasma, combining the cellular components with the platelet-poor plasma to form a first mixture, and separating the first mixture into mononuclear cells and at least one component.

Described is a method for controlling fluid volume balance. A controller is configured with a first set of inputs comprising a hematocrit, a total blood volume, and an ACD ratio. A maximum extracorporeal RBC amount during the procedure is estimated based on the first set of inputs. A fluid circuit is primed with a priming fluid. Whole blood is drawn from a blood source and separated into a RBC component, a target cell component, and a plasma component. The target cell component is directed to a product container. The product container comprising the target cell component is treated. A treated target cell component, a portion of the RBC component remaining in the fluid circuit, and/or a portion of the plasma component remaining in the fluid circuit are returned to the blood source. A first response action is provided if the maximum extracorporeal RBC amount estimated is above a programmed limit.

Described is a method for controlling fluid volume balance. A controller is configured with a first set of inputs comprising a hematocrit, a total blood volume, and an ACD ratio. A maximum extracorporeal RBC amount during the procedure is estimated based on the first set of inputs. A fluid circuit is primed with a priming fluid. Whole blood is drawn from a blood source and separated into a RBC component, a target cell component, and a plasma component. The target cell component is directed to a product container. The product container comprising the target cell component is treated. A treated target cell component, a portion of the RBC component remaining in the fluid circuit, and/or a portion of the plasma component remaining in the fluid circuit are returned to the blood source. A first response action is provided if the maximum extracorporeal RBC amount estimated is above a programmed limit.

A blood separation system is provided that includes a blood separation device that includes a centrifugal separator and a spinning membrane separator drive unit incorporated into a common case and a fluid flow circuit having both a separation chamber configured to be mounted in the centrifugal separator of the blood separation device and a spinning membrane separator configured to be received in the spinning membrane separator drive unit. In an exemplary procedure, the system is used to collect concentrated platelets and/or concentrated platelets and plasma, and to further permit harvesting of the mononuclear cells from the centrifugal separator at the conclusion of platelet collection, and transfer of the mononuclear cells to the spinning membrane separator.

Blood in a separation chamber is separated into a red blood cell layer, a plasma constituent, and a mononuclear cell-containing layer. A portion of the plasma constituent exits the chamber via a plasma outlet, while a first portion of the red blood cell layer exits via a red blood cell outlet. A second portion of the red blood cell layer exits the chamber via the red blood cell outlet and is collected. At least a portion of the collected red blood cell layer may then be conveyed to the chamber via the red blood cell outlet to convey at least a portion of the mononuclear cell-containing layer out of the chamber via the plasma outlet for collection. A second portion of the plasma constituent may be conveyed out of the chamber via the plasma outlet to more fully collect the mononuclear cell-containing layer without the use of collected plasma.

A method for separating cells in a biofluid includes pretreating the biofluid by introducing an additive, flowing the pretreated biofluid through a microfluidic separation channel, and applying acoustic energy to the microfluidic separation channel. A system for microfluidic cell separation, capable of separating target cells from non-target cells in a biofluid includes at least one microfluidic separation channel, a source of biofluid, a source of additive, and at least one acoustic transducer coupled to the microfluidic separation channel. A kit for microfluidic cell separation includes a microfluidic separation channel connected to an acoustic transducer, a source of an additive, and instructions for use.