Cell concentration devices and methods that include an insert defining a lumen and a cannula assembly

Abstract

A system and associated method for concentrating and separating components of different densities from fluid containing cells using a centrifuge includes a container defining a cavity for receiving the fluid. The container has a top, a sidewall extending from the top, and a bottom disposed opposite the top and in sealing engagement with the sidewall. An insert is slidably disposed in the cavity of the container and defines a lumen through the insert. The lumen, which includes a hole and a funnel-shaped upper portion in fluid communication with the hole, forms an open fluid path between opposite ends of the insert. The insert has a density such that upon centrifugation a selected component of the fluid resides within the lumen. A container port is disposed in the top of the container to transfer the fluid into the container and to withdraw a fluid component other than the selected component from the container. The system includes a manifold that includes a manifold port, a vent to vent the container, and a connector to couple to the container port. A cannula is receivable in the manifold port and extendable through the container port into the container and into the lumen of the insert to withdraw the selected component from the lumen.

1 . A system for separating components of different densities from a fluid containing cells using a centrifuge, the system comprising:
 a container having a top, a sidewall extending from the top, and a bottom disposed opposite the top and in sealing engagement with the sidewall, the container defining a cavity for receiving the fluid;   an insert slidably disposed in the cavity and defining a lumen through the insert, the lumen including a hole and a funnel-shaped upper portion in fluid communication with the hole, the insert having a density such that upon centrifugation a selected component of the fluid resides within the lumen, the lumen forming an open fluid path between opposite ends of the insert;   a container port disposed in the top of the container to transfer the fluid into the container and to withdraw a fluid component other than the selected component;   a manifold including a manifold port, a vent to vent the container, and a connector to couple to the container port; and   a cannula receivable in the manifold port and extendable through the container port into the container and into the lumen of the insert to withdraw the selected component from the lumen.   
     
     
         2 . The system of  claim 1 , wherein the cannula includes a closed end to close the hole in the insert and a side port to withdraw the selected component, the cannula and the insert forming a seal when the closed end of the cannula closes off the hole in the insert. 
     
     
         3 . The system of  claim 1 , wherein the cannula is a first cannula, and further including a second cannula extendable through the container port to withdraw the component other than the selected component. 
     
     
         4 . The system of  claim 3 , wherein the second cannula is receivable in the manifold port. 
     
     
         5 . The system of  claim 1 , wherein the container is a syringe and the bottom is a plunger. 
     
     
         6 . The system of  claim 5 , wherein the plunger has a removable handle. 
     
     
         7 . The system of  claim 5 , wherein the plunger is a first plunger and further including a second plunger disposed in the syringe below the first plunger to move the first plunger. 
     
     
         8 . The system of  claim 1 , further including a clamping mechanism to hold the insert in place after centrifugation, the clamping mechanism configured to press the sidewall inward against the insert. 
     
     
         9 . The system of  claim 1 , wherein the selected component is buffy coat and the component other than the selected component is blood plasma. 
     
     
         10 . The system of  claim 1 , wherein the insert is rigid. 
     
     
         11 . The system of  claim 1 , wherein the volume contained in the lumen upper portion of the insert is between 5% and 20% of the volume of the container cavity. 
     
     
         12 . A method of separating components of different densities from a fluid containing cells using a centrifuge, the method comprising:
 receiving the fluid in a separation system, the system comprising
 a container having a top, a sidewall extending from the top, and a bottom disposed opposite the top and in sealing engagement with the sidewall, the container defining a cavity for receiving the fluid, 
 an insert slidably disposed in the cavity and defining a lumen through the insert, the lumen including a hole and a funnel-shaped upper portion in communication with the hole, the insert having a density such that upon centrifugation a selected component of the fluid resides within the lumen of the insert, the lumen forming an open fluid path between opposite ends of the insert, and 
 a container port disposed in the top of the container; 
   applying centrifugal force to the separation system;   after centrifugation, withdrawing a fluid component other than the selected component through the container port;   coupling a manifold to the container port, the manifold including a manifold port and a vent to vent the container; and   extending a cannula through the container port into the container and into the lumen of the insert, the cannula receivable in the manifold port; and   withdrawing the selected component with the cannula from the lumen of the insert.   
     
     
         13 . The method of  claim 12 , further comprising closing the hole in the insert with a closed end of the cannula. 
     
     
         14 . The method of  claim 13 , wherein closing the hole in the insert comprises forming a seal between the cannula and the insert. 
     
     
         15 . The method of  claim 12 , wherein the container is a syringe and the bottom is a plunger. 
     
     
         16 . The method of  claim 15 , wherein the plunger is a first plunger, and further including moving the first plunger with a second plunger disposed in the syringe below the first plunger to transfer the fluid into the container. 
     
     
         17 . The method of  claim 12 , wherein withdrawing the selected component includes withdrawing the selected component through a side port in the cannula. 
     
     
         18 . The method of  claim 12 , wherein the cannula is a first cannula and wherein withdrawing the component other than the selected component includes extending a second cannula through the container port, the second cannula receivable in the manifold port, and withdrawing the component other than the selected component with the second cannula. 
     
     
         19 . The method of  claim 12 , wherein the manifold is coupled to the container before the withdrawing of the fluid component other than the selected component. 
     
     
         20 . The method of  claim 12 , further comprising, with a clamping mechanism, holding the insert in place after centrifugation. 
     
     
         21 . A method of separating components of different densities from a fluid containing cells using a centrifuge, the method comprising:
 receiving the fluid in a separation system, the system comprising
 a container, having a bottom, a top disposed opposite the bottom, and a sidewall extending from the top, the container defining a cavity for receiving the fluid, 
 an insert slidably disposed in the cavity, the insert including a funnel-shaped upper portion and a hole therethrough, the insert having a density such that upon centrifugation a selected component of the fluid resides within the upper portion of the insert, and 
 a container port disposed in the top of the container; 
   applying centrifugal force to the system;   inserting a cannula into the container through the container port to butt against the insert, the cannula having one or more side ports displaced from a distal end of the cannula;   withdrawing the selected component through the side ports in the cannula;   ejecting at least a portion of the withdrawn component through the side ports causing one or more fluid jets in the funnel-shape upper portion of the insert to release cells that adhere to the insert; and   withdrawing the ejected portion and cells released by the fluid jets through the side ports.   
     
     
         22 . A method of separating components of different densities from a fluid containing cells using a centrifuge, the method comprising:
 receiving the fluid in a container having a top, a sidewall extending from the top, and a bottom disposed opposite the top and in sealing engagement with the sidewall, the container defining a cavity for receiving the fluid;   applying centrifugal force to the container;   after centrifugation, inserting a cannula through an elastomeric membrane and into the container;   withdrawing a selected component of the fluid through the cannula; and   with a vent, allowing air through the elastomeric membrane and into the container to displace fluid withdrawn through the cannula.   
     
     
         23 . The method of  claim 22 , wherein the container includes an insert slidably disposed in the cavity, the insert, upon centrifugation, allowing fluid components of a density lower than a certain density to rise above the insert and fluid components of a density higher than the certain density to sink below the insert. 
     
     
         24 . The method of  claim 23 , wherein the insert includes a funnel-shaped upper portion and a hole therethrough, the insert having a density such that upon centrifugation the selected component of the fluid resides within the upper portion of the insert. 
     
     
         25 . The method of  claim 22 , further comprising coupling a manifold to the container, the manifold including the elastomeric membrane and the vent.