Therapeutic cell washing, concentration, and separation utilizing acoustophoresis

Abstract

Multi-stage acoustophoretic devices for continuously separating a second fluid or a particulate from a host fluid are disclosed. Methods of operating the multi-stage acoustophoretic devices are also disclosed. The systems may include multiple acoustophoretic devices fluidly connected to one another in series, each acoustophoretic device comprising a flow chamber, an ultrasonic transducer capable of creating a multi-dimensional acoustic standing wave, and a reflector. The systems can further include pumps and flowmeters.

1 . A method exchanging media, the method comprising:
 providing an initial mixture of a first media and particles to a chamber of an acoustophoretic device, the acoustophoretic device including at least one ultrasonic transducer that includes a piezoelectric material;   driving the at least one ultrasonic transducer to create an acoustic wave an acoustic region in the chamber;   providing the initial mixture to the acoustic region, such that at least a portion of the particles are trapped and held against fluid flow in the acoustic region;   forming clusters of the trapped particles to grow in size in the acoustic region; and   flowing a second media to the chamber while the particles and clusters are retained in the chamber to cause the first media to flow out of the chamber;   wherein an input rate of one or more of the initial mixture or the second media to the chamber is in a range of from about 10 ml/min to about 15 ml/min.   
     
     
         2 . The method of  claim 1 , wherein the second media is a biocompatible wash or a buffer solution. 
     
     
         3 . The method of  claim 1 , wherein the particles are cells. 
     
     
         4 . The method of  claim 1 , wherein the particles are microcarrier/cell complexes. 
     
     
         5 . The method of  claim 1 , wherein the initial mixture has a density of about 0.5 million particles/ml to about 5 million particles/ml. 
     
     
         6 . The method of  claim 1 , further comprising concentrating the particles in the initial mixture. 
     
     
         7 . The method of  claim 6 , further comprising concentrating the particles to a concentrate volume that is about 25 to about 50 times less than a volume of the initial mixture. 
     
     
         8 . The method of  claim 7 , further comprising concentrating the particles in the initial mixture to a concentrated particle density of about 25 to about 50 times greater than a particle density of the initial mixture. 
     
     
         9 . The method of  claim 1 , wherein a cell density of the first media output from the chamber is about 0.0 to about 0.5 million cells/ml. 
     
     
         10 . The method of  claim 9 , wherein the first media output is from a concentrate process and a wash process. 
     
     
         11 . The method of  claim 1 , further comprising conducting a spectrophotometer process on the chamber to determine wash efficacy. 
     
     
         12 . A method of recovering cells from a cell culture, comprising:
 feeding an initial mixture of the cell culture to a flow chamber of an acoustophoretic device, the acoustophoretic device including at least one ultrasonic transducer that includes a piezoelectric material that is configured to be driven to generate a multi-dimensional acoustic wave in the flow chamber; and   driving the at least one ultrasonic transducer to generate a multi-dimensional acoustic wave in an acoustic region in the flow chamber;   providing the initial mixture to the acoustic region; and   retaining the cells from the initial mixture in the acoustic region to form clusters of the cells to grow in size in the acoustic region;   wherein an input rate of the initial mixture to the flow chamber is in a range of from about 10 ml/min to about 15 ml/min.   
     
     
         13 . The method of  claim 12 , wherein the cell density of the concentrated cells is about 25 to about 50 times greater than the cell density of the initial mixture. 
     
     
         14 . The method of  claim 12 , wherein a volume of the concentrated cells is 25 to about 50 times less than a volume of the initial mixture. 
     
     
         15 . The method of  claim 12 , wherein the concentrated cells are obtained in about 35 minutes or less. 
     
     
         16 . The method of  claim 12 , further comprising washing the concentrated cells, wherein a cell density of a wash output of the flow chamber is about 0.0 to about 0.5 million cells/ml. 
     
     
         17 . An acoustophoretic device, comprising:
 a flow chamber with a first outlet;   at least one ultrasonic transducer coupled to the flow chamber and including a piezoelectric material that is adapted to be driven to generate an acoustic wave in an acoustic region of the flow chamber, such that particles are trapped to form particle clusters in the acoustic region that can grow in size when a fluid and particle mixture is provided to the acoustic region;   a diminished particle concentration region adjacent the acoustic region and in fluid communication with the first outlet, the diminished particle concentration region being interposed between the acoustic region and the first outlet; and   a fluid input region on an opposite side of the acoustic region from the first outlet that permits fluid flow into the acoustic region, such that an input fluid flows through the acoustic region while the particles and particle clusters remain trapped in the acoustic region;   wherein the dimensions of the flow chamber and the first outlet are sized to accommodate a flow rate in a range of from about 10 ml/min to about 15 ml/min.   
     
     
         18 . The acoustophoretic device of  claim 17 , wherein the flow chamber can contain a cell capacity of about 4 billion to about 40 billion cells.