Normothermic, hypothermic and cryopreservation maintenance and storage of cells, tissues and organs in gel-based media

Abstract

Gel-based medium compositions and a method of use thereof in normothermic, hypothermic or cryopreservative storage and transport of cell samples are described. These gel-based compositions contain a cell maintenance and preservation medium together with a gelling agent. Such gel-based medium compositions protect various cell samples, such as animal or plant organs, tissues and cells, from the mechanical, physiological and biochemical stresses inherently associated with liquid preservation techniques.

What is claimed is:  
     
         1 . Gel-based medium compositions for transport or storage of cell samples, the gel-based compositions comprising at least one cell maintenance and preservation medium and at least one gelling agent.  
     
     
         2 . The gel-based medium compositions according to  claim 1 , wherein transport or storage of cells occurs at a temperature selected from the group consisting of normothermic, hypothermic and cryopreservative temperatures.  
     
     
         3 . The gel-based medium compositions according to  claim 2 , wherein transport or storage of cell samples occurs at a temperature ranging from about −196° C. to about 37° C.  
     
     
         4 . The gel-based medium compositions according to  claim 1 , wherein the cell samples are obtained from a source selected from the group consisting of plants, animals, fungi, and microbes.  
     
     
         5 . The gel-based medium compositions according to  claim 4 , wherein the cell samples are selected from the group consisting of organs, tissues, cell monolayers and single cells.  
     
     
         6 . The gel-based medium compositions according to  claim 5 , wherein the cell samples are obtained from humans.  
     
     
         7 . The gel-based medium compositions according to  claim 1 , wherein the at least one cell maintenance and preservation medium is liquid.  
     
     
         8 . The gel-based medium compositions according to  claim 1 , wherein the at least one gelling agent is a member selected from the group consisting of gelatin, carrageenan, agarose, collagen and plant-based gelling agents.  
     
     
         9 . A gel-based medium composition for transport or storage of cell samples, the composition comprising: 
 (a) one or more electrolytes selected from the group consisting of potassium ions at a concentration ranging from about 10-145 mM, sodium ions ranging from about 10-120 mM, magnesium ions ranging from about 0.1-10 mM, and calcium ions ranging from about 0.01-1.0 mM;    (b) a macromolecular oncotic agent having a size sufficiently large to limit escape from the circulation system and effective to maintain oncotic pressure equivalent to that of blood plasma and selected from the group consisting of human serum albumin, polysaccharide and colloidal starch;    (c) a biological pH buffer effective under physiological and hypothermic conditions;    (d) a nutritive effective amount of at least one simple sugar;    (e) an impermeant and hydroxyl radical scavenging effective amount of mannitol;    (f) an impermeant anion impermeable to cell membranes and effective to counteract cell swelling during cold exposure, said impermeant ion being at least one member selected from the group consisting of lactobionate, gluconate, citrate and glycerophosphate-like compounds;    (g) a substrate effective for the regeneration of ATP, said substrate being at least one member selected from the group consisting of adenosine, fructose, ribose and adenine;    (h) at least one agent which regulates cellular levels of free radicals; and    (i) at least one gelling agent.    
     
     
         10 . The gel-based medium composition according to  claim 9 , wherein the gelling agent is a member selected from the group consisting of gelatin, carrageenan, agarose, collagen and plant-based gelling agents.  
     
     
         11 . The gel-based medium composition according to  claim 9 , wherein the transport or storage of cell samples occurs at a temperature ranging from about −196° C. to about 37° C.  
     
     
         12 . The gel-based medium composition according to  claim 9 , wherein the cell samples are obtained from a source selected from the group consisting of plants, animals, fungi, and microbes.  
     
     
         13 . The gel-based medium composition according to  claim 12 , wherein the source of the cell samples is a fetal, neonatal, juvenile or adult animal.  
     
     
         14 . The gel-based medium composition according to  claim 13 , wherein the cell samples are obtained from humans.  
     
     
         15 . A method of storing cell samples in a gel-based medium composition, the method comprising: 
 warming a gel-based medium composition to melt a gelling agent contained therein;    suspending cell samples in a cell preservation solution without a gelling agent;    mixing the suspended cell samples with the warmed gel-based medium composition;    cooling the cell samples to solidify the gelling agent; and    transferring the cooled cell samples to a desired storage temperature.    
     
     
         16 . The method according to  claim 15  further comprising optionally transporting the cooled cell samples in a gelled state.  
     
     
         17 . The method according to  claim 16 , wherein the transport or storage of cell samples occurs at a temperature ranging from about −196° C. to about 37° C.