Cell Freezing Kits and Their Use Thereof

Abstract

The disclose provides a cell freezing kit having a device equipped with a filter, and a container having hermetic closing means, which is capable of containing the device equipped with a filter, and methods of use thereof.

1 - 22 . (canceled) 
     
     
         23 . A cell freezing kit, comprising:
 (i) a device equipped with a filter; and   (ii) a container having hermetic closing means, which is capable of containing the device equipped with a filter,   
       wherein the filter has hydrophobic properties and/or the kit also comprises a hydrophobic solution. 
     
     
         24 . The cell freezing kit of  claim 23 , wherein the average diameter of the pores of the filter is between 2 and 10 microns. 
     
     
         25 . The cell freezing kit of  claim 23 , wherein the filter has hydrophobic properties so that the freezing medium does not pass through the filter in the absence of a driving force greater than gravitational force. 
     
     
         26 . The cell freezing kit of  claim 23 , which comprises a hydrophobic solution capable of passing through the filter only when a driving force greater than gravitational force is applied thereto. 
     
     
         27 . The cell freezing kit of  claim 26 , wherein the hydrophobic solution is a solution of glycerol. 
     
     
         28 . The cell freezing kit of  claim 26 , wherein the hydrophobic solution is a sterile oil free of endotoxins. 
     
     
         29 . The cell freezing kit of  claim 23 , wherein the device equipped with a filter is a centrifugal filter. 
     
     
         30 . The cell freezing kit of  claim 23 , wherein the hermetic closing means of the container comprise a screw cap. 
     
     
         31 . The cell freezing kit of  claim 23 , wherein the assembly of the container and the device equipped with a filter constitutes a centrifugation unit suitable for centrifugation between 50 and 1500 g. 
     
     
         32 . The cell freezing kit of  claim 31 , which comprises one or several centrifugation units, wherein said centrifugation units are sterile or sterilizable. 
     
     
         33 . A method for freezing cells using the device of  claim 23 , comprising:
 (i) introducing cells in a freezing medium in the device equipped with a filter;   (ii) putting the device equipped with a filter into the container;   (iii) hermetically closing the container; and   (iv) placing the container at a freezing temperature.   
     
     
         34 . The method of  claim 33 , wherein step (ii) is performed before step (i). 
     
     
         35 . The method of  claim 33 , wherein a drop of a hydrophobic solution is added to the filter before step (i). 
     
     
         36 . The method of  claim 33 , wherein the device equipped with a filter and the container are sterilized before step (i). 
     
     
         37 . The method of  claim 33 , wherein the amount of cells placed in each device equipped with a filter is between 10 4  and 10 7 . 
     
     
         38 . A method for thawing cells frozen using the device of  claim 23 , comprising:
 (i) placing the container, containing a frozen cell suspension in a device equipped with a filter, at a temperature which allows the cell suspension to thaw;   (ii) applying to the cell suspension a force which drives the passing of the freezing medium through the filter;   (iii) resuspending the cells in cell medium.   
     
     
         39 . The method of  claim 38 , wherein step (ii) is performed by centrifugation. 
     
     
         40 . The method of  claim 39 , wherein the centrifugation is performed during less than or equal to 10 minutes, at a speed which makes it possible to apply to the cell suspension an acceleration of between 50 and 1500 g, preferably between 100 and 200 g.