Cell suspension preparation technique and device

Abstract

The present invention provides a unique method and or device suitable for producing a transplantable cellular suspension of living tissue suitable for grafting to a patient. In applying the method and or in using the device donor tissue is harvested, subjected to a cell dissociation treatment, cells suitable for grafting back to a patient are collected and dispersed in a solution that is suitable for immediate dispersion over the recipient graft site.

What is claimed is:  
     
         1 . A method for preparing a cell suspension suitable for application to a patient, which method comprises the steps of: 
 (a) subjecting a tissue sample including cells suitable for grafting to a patient, to at least a physical and or chemical dissociating means capable of dissociating cellular stratum in the tissue sample;    (b) removing the tissue sample from the dissociating means used in step (a) and harvesting in the presence of a nutrient solution cells from the tissue sample, cells suitable for grafting on to a patient wherein the nutrient solution is (i) free of xenogenic serum, (ii) capable of maintaining the viability of the cells until applied to a patient and (iii) is suitable for direct application to a region on a patient undergoing tissue grafting; and    (c) filtering the cellular suspension produced according to step (b) to remove large cellular conglomerates.    
     
     
         2 . A method according to  claim 1  wherein the enzyme suitable for dissociating cohesive pieces of tissue stratum in the sample is trypsin or a trypsin-like enzyme.  
     
     
         3 . A method according to  claim 2  wherein the enzyme is selected from the group consisting of trypsin, trypsin-EDTA, dispase, collagenase, thermolysin, pronase, hyaluronidase, pancreatin, elastase and papain.  
     
     
         4 . A method according to  claim 1  wherein the nutrient solution is Hartmann's solution.  
     
     
         5 . A cell suspension produced according to the method of  claim 1 .  
     
     
         6 . A cell suspension according to  claim 5  prepared from autologous cells.  
     
     
         7 . A method of treating a patient in need of graft surgery, said method comprising the steps of: 
 (a) preparing a cell suspension according to the method of claim 1 ; and    (b) administering the suspension directly to a region on the patient that requires a cell graft in a manner that facilitates spreading of the cell suspension in a relatively even distribution over the graft region.    
     
     
         8 . Use of a cellular suspension suitable for grafts, which suspension is prepared according to the following steps: 
 (a) subjecting a tissue sample including cells suitable for grafting to a patient, to an enzyme suitable for dissociating cohesive pieces of the tissue stratum in the sample;    (b) removing the sample from the enzyme solution used in step (a) and harvesting in the presence of a nutrient solution cells from the tissue sample, which cells are suitable for grafting on to a patient wherein the nutrient solution is (i) free of xenogenic serum, (ii) capable of maintaining the viability of the cells until applied to a patient and (iii) is suitable for direct application to a region on a patient undergoing tissue grafting; and    (c) filtering the cellular suspension produced according to step (b) to remove large cellular conglomerates;    for the preparation of therapeutic preparation suitable for the treatment of tissue disorders requiring grafting.    
     
     
         9 . The use according to  claim 8  wherein the nutrient solution is Hartmann's solution.  
     
     
         10 . An apparatus for developing a tissue regeneration solution, said apparatus comprising: 
 (a) a heating means suitable for heating an enzyme solution to a required temperature and which is capable of maintaining that solution at the desired temperature for a suitable amount of time; and    (b) a filter recess comprising a filter means capable of separating large cellular congregates from a cellular suspension.    
     
     
         11 . The apparatus according to  claim 10 , which additionally comprises a reservoir capable of holding a tissue sample and a nutrient solution.  
     
     
         12 . The apparatus according to  claim 10 , which includes one or more fluid containment wells for storage of fluids.  
     
     
         13 . An apparatus for developing a tissue regeneration solution, comprising a first and second member wherein: 
 (i) the first member includes: 
 (a) a heating means suitable for heating an enzyme solution to a required temperature and which is capable of maintaining that solution at the desired temperature for a suitable amount of time;  
 (b) a filter recess comprising a filter means capable of separating large cellular congregates from a cellular suspension;  
 (c) at least a fluid containment well for storage of nutrient solution;  
   (ii) the second member forms a reservoir capable of withholding a tissue sample and nutrient solution in fluid containment; and    wherein the first member provides a seat upon which the second member may be placed during manipulation of the tissue.