Normothermic, hypothermic and cryopreservation maintenance and storage of cells, tissues and organs in gel-based media
AssigneeBioLife Solutions, Inc.
InventorsJohn M. Baust, Robert Van Buskirk, John G. Baust
Life of this patent
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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.
Claims17
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.
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.
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.
Dependencies are inferred from the claim text.