WO2020242970A1 - Preservation using sugars - Google Patents

Abstract

Embodiments of the present invention relate to methods of preparing a cell, tissue, organ, plant, or inanimate biomaterial for preservation, wherein the method includes contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition including sucrose, sucralose, trehalose, and/or maltose.

Description

PRESERVATION USING SUGARS

Statement of Priority

This application claims the benefit, under 35 U.S.C. § 119(e), of U.S. Provisional Application Serial No. 62/852,600, filed May 24, 2019, the entire contents of which are incorporated by reference herein.

Reservation of Copyright

A portion of the disclosure of this patent document contains material which is subject to copyright protection. The copyright owner, East Carolina University, Greenville, North Carolina, a constituent institution of the University of North Carolina, has no objection to the reproduction by anyone of the patent document or the patent disclosure, as it appears in Patent Office patent files or records, but otherwise reserves all copyright rights whatsoever.

Field of the Invention

The present invention relates to methods and compositions useful for preservation of cells, tissues, organs, plants, and inanimate biomaterial.

Background

Typically, preservation without adding some protective chemical will result in degradation. For example, in cryopreservation the cause of death is generally ice crystal formation, which punctures cells leading to cell death. The techniques for preservation currently being used may employ toxic chemicals which must be removed from the cells before they are used or transfused. Glycerol, a common cryopreservative used for storing frozen red blood cells (RBCs), must be removed using specialized equipment before transfusion of the cells into patients. Dimethylsulfoxide (DMSO), another common cryopreservative, has limited use due to adverse effects on many cell lines.

The present invention overcomes previous shortcomings in the art by providing preservation methods with nontoxic compounds.

Summary

One aspect of the present invention provides methods of preparing a cell, tissue, organ or plant for preservation, comprising contacting the cell, tissue, organ or plant with a composition comprising from about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose; and subjecting the cell, tissue organ or plant to a temperature from about - 30°C to about 20°C.

Another aspect of the present invention provides methods of reducing cell death during preservation of a cell, tissue, organ or plant, comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C.

Another aspect of the present invention comprises methods of preserving cell function during preservation of a cell, tissue, organ or plant, comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C, wherein the cell function is reduced by less than about 50% ( e.g ., by less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50%, etc.) as compared to a control.

Another aspect of the present invention provides methods of reducing bacterial contamination during preservation of a cell, tissue, organ or plant, comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C, wherein the bacterial contamination is reduced by more than 50% (e.g., more than about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, or 100%) as compared to a control.

A further aspect of the present invention provides methods for the preservation of an inanimate biomaterial, comprising: (a) contacting the inanimate biomaterial with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose; and (b) subjecting the inanimate biomaterial to a temperature from about -80°C to about 50°C.

Another aspect of the present invention provides methods of reducing bacterial contamination during preservation of an inanimate biomaterial, comprising contacting the inanimate biomaterial with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the inanimate biomaterial to a temperature from about -80°C to about 50°C, wherein the bacterial contamination is reduced by more than 50% (e.g, more than about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, or 100%) as compared to a control. An additional aspect of the present invention provides methods for lyopreservation of liposomes, comprising (a) contacting a liposome comprising a drug with a composition comprising sucrose, sucralose, trehalose and/or maltose; and (b) subjecting the liposome to a temperature from about -30°C to about 20°C.

Further aspects of the present invention include the use of sucrose, sucralose, trehalose and/or maltose for the preparation of pharmaceutical compositions, transfusable preparations and/or transplantable preparations for the prevention and/or treatment of diseases and disorders for human and veterinary purposes, as well as preservation media comprising sucrose, sucralose, trehalose, and/or maltose in a suitable carrier for preservation, preserved preparations of cells, tissues, organs or plants or inanimate biomaterial prepared by the methods of the present invention; pharmaceutical compositions comprising a cell subjected to a method of the present invention; and kits comprising preserved cells, tissues, organs, plants, or inanimate biomaterial as described herein or components to effectuate preservation of cells, tissues, organs, plants, or inanimate biomaterial.

These and other aspects of the invention are addressed in more detail in the description of the invention set forth below.

Brief Description of the Drawings

FIGS. 1A-1B show hemolysis data of red blood cells treated with sucralose (FIG.

1A) or sucrose (FIG. IB). The graphs show percent hemolysis of blood treated with 0.00% sucralose (squares), 0.50% sucralose (circles) and 1.00% sucralose (triangles). Error bars indicate standard error on the mean (n=3).

FIGS. 2A-2B show two exemplar experiments of the % hemolysis of sheep red blood cells treated with sucralose in varying concentrations, as a function of storage time in a refrigerator. FIG. 2A shows percent hemolysis of blood treated with sucralose at various concentrations (%w/v) indicated on the graph. Error bars indicate standard error on the mean (n=3). FIG. 2B shows percent hemolysis of blood treated with sucrose at various

concentrations (%w/v) indicated on the graph. Error bars indicate standard error on the mean (n=3).

FIG. 3 shows that increasing concentrations of sucralose reduce sheep’s blood hemolysis over the course of 70 days at 4°C.

FIG. 4 shows that AS1 in combination with sucralose demonstrate a synergistic effect in reducing relative hemolysis in sheep’s blood past 20 days at 4°C. Relative absorbance is displayed, which is directly related to percent hemolysis. Detailed Description

This invention may be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

The terminology used in the description herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used in the description and the appended claims, the singular forms“a,”“an” and“the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.

Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the present application and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. The terminology used in the description herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

All patent, patent application references and other references referred to in this patent application are incorporated by reference herein in their entirety as if set forth fully herein.

Also as used herein,“and/or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of

combinations when interpreted in the alternative (“or”).

The term "about," as used herein when referring to a measurable value such as an amount of dose ( e.g an amount of a fatty acid) and the like, is meant in some embodiments to encompass variations of ± 20%, ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of the specified amount or value.

Unless the context indicates otherwise, it is specifically intended that the various features of the embodiments of the invention described herein may be used in any

combination. For example, features described in relation to one embodiment may also be applicable to and combinable with other embodiments and aspects of the invention.

Moreover, the embodiments of the present invention also contemplate that in some embodiments, any feature or combination of features set forth herein may be excluded or omitted. To illustrate, if the specification states that a complex comprises components A, B and C, in some embodiments, any of A, B or C, or a combination thereof, may be omitted and disclaimed.

The term“comprise,”“comprises” and“comprising” as used herein, specify the presence of the stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.

As used herein, the transitional phrase“consisting essentially of’ (and grammatical variants) is to be interpreted as encompassing the recited materials or steps“and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. See, In re Herz , 537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463 (CCPA 1976) (emphasis in the original); see also MPEP § 2111.03. Thus, the term“consisting essentially of’ as used herein should not be interpreted as equivalent to“comprising.”

“Preservation” refers to a process whereby cells, tissues, organs, or any other substances susceptible to damage caused by chemical reactivity or time ( e.g ., inanimate biomaterial) can be preserved by the methods disclosed herein. In some embodiments, preservation may be preserved by cooling to sub-zero Celsius temperatures (e.g.,

cryopreservation).

“Cryopreservation” or“cryoconservation” refers to a process whereby cells, tissues, organs or any other substances susceptible to damage caused by chemical reactivity or time can be preserved by cooling to sub-zero Celsius temperatures.

Cryogenic preservation of cells in suspension is a well-established and accepted technique for long term archival storage and recovery of live cells. As a general method, cells may be suspended in a cryopreservation media typically including salt solutions, buffers, nutrients, growth factors, proteins, and cryopreservatives. The cells are then distributed to archival storage containers of the desired size and volume, and the containers are then reduced in temperature until the container contents are frozen. Generally, long-term archival conditions may include liquid nitrogen gas storage where temperatures can be approximately -190°C.

The recovery of live cells and suitable tissues, organs and plants preserved by such methods is largely dependent upon minimizing injurious ice crystal growth in the intracellular region during both the freezing and thawing processes. A combination of two methods for reducing intracellular ice crystal growth is typically practiced in the freezing process. A cryoprotectant to permeate the cell membrane (or plant cell wall) and inhibit ice crystal nucleation and growth both extracellularly and intracellularly is desired as well as managing the reduction in sample temperature over time.

“Vitrification” refers to zero ice formation.

“Freezing” refers to a phase transition from which a liquid or semi-solid becomes a solid when its temperature is lowered to or below its freezing point.

“Crystallization” refers to the formation of a crystalline solid from or within a uniform liquid or semi-liquid resulting from the removal of heat.

“Subjects” as used herein are generally human subjects and include, but are not limited to,“patients.” The subjects may be male or female and may be of any race or ethnicity, including, but not limited to, Caucasian, African-American, African, Asian, Hispanic, Indian, etc. The subjects may be of any age, including newborn, neonate, infant, child, juvenile, adolescent, adult, and geriatric. Subjects may also include animal subjects, particularly mammalian subjects such as canines, felines, bovines, caprines, equines, ovines, porcines, rodents ( e.g ., rats and mice), lagomorphs, primates (including non-human primates), etc., for prevention and treatment purposes as well as veterinary medicine and/or pharmaceutical drug development purposes.

The term“cells” as used herein, refer to prokaryotic and eukaryotic cells. The cells used in accordance with the methods described herein may include primary cells that have been isolated from a tissue or organ, using one or more art-known proteases, e.g., collagenase, dispase, trypsin, or the like. The cells may be embryonic stem cells, embryonic germ cells, induced pluripotent stem cells, mesenchymal stem cells, bone marrow-derived mesenchymal stem cells (BM-MSCs), tissue plastic-adherent placental stem cells (PDACs), umbilical cord stem cells, amniotic fluid stem cells, amnion derived adherent cells

(AMDACs), osteogenic placental adherent cells (OPACs), adipose stem cells, limbal stem cells, dental pulp stem cells, myoblasts, endothelial progenitor cells, neuronal stem cells, exfoliated teeth derived stem cells, hair follicle stem cells, dermal stem cells,

parthenogenically derived stem cells, reprogrammed stem cells, amnion derived adherent cells, or side population stem cells. The cells used herein may also include blast cells, cloned cells, fertilized ova, placental cells, keratinocytes, basal epidermal cells, hair shaft cells, hair- root sheath cells, surface epithelial cells, basal epithelial cells, urinary epithelial cells, salivary gland cells, mucous cells, serous cells, von Ebner's gland cells, mammary gland cells, lacrimal gland cells, ceruminous gland cells, eccrine sweat gland cells, apocrine sweat gland cells, Moll gland cells, sebaceous gland cells, Bowman's gland cells, Brunner's gland cells, seminal vesicle cells, prostate gland cells, bulbourethral gland cells, Bartholin's gland cells, Littre gland cells, uterine endometrial cells, goblet cells of the respiratory or digestive tracts, mucous cells of the stomach, zymogenic cells of the gastric gland, oxyntic cells of the gastric gland, insulin-producing b cells, glucagon-producing a cells, somatostatin-producing D cells, pancreatic polypeptide-producing cells, pancreatic ductal cells, Paneth cells of the small intestine, type II pneumocytes of the lung, Clara cells of the lung, anterior pituitary cells, intermediate pituitary cells, posterior pituitary cells, hormone secreting cells of the gut or respiratory tract, thyroid gland cells, parathyroid gland cells, adrenal gland cells, gonad cells, juxtaglomerular cells of the kidney, macula densa cells of the kidney, peripolar cells of the kidney, mesangial cells of the kidney, brush border cells of the intestine, striated duct cells of exocrine glands, gall bladder epithelial cells, brush border cells of the proximal tubule of the kidney, distal tubule cells of the kidney, nonciliated cells of ductulus efferens, epididymal principal cells, epididymal basal cells, hepatocytes, fat cells, type I pneumocytes, pancreatic duct cells, nonstriated duct cells of the sweat gland, nonstriated duct cells of the salivary gland, nonstriated duct cells of the mammary gland, parietal cells of the kidney glomerulus, podocytes of the kidney glomerulus, cells of the thin segment of the loop of Henle, collecting duct cells, duct cells of the seminal vesicle, duct cells of the prostate gland, vascular endothelial cells, synovial cells, serosal cells, squamous cells lining the

perilymphatic space of the ear, cells lining the endolymphatic space of the ear, choroids plexus cells, squamous cells of the pia-arachnoid, ciliary epithelial cells of the eye, corneal endothelial cells, ciliated cells having propulsive function, ameloblasts, planum semilunatum cells of the vestibular apparatus of the ear, interdental cells of the organ of Corti, fibroblasts, pericytes of blood capillaries, nucleus pulposus cells of the intervertebral disc, cementoblasts, cementocytes, odontoblasts, odontocytes, chondrocytes, osteoblasts, osteocytes,

osteoprogenitor cells, hyalocytes of the vitreous body of the eye, stellate cells of the perilymphatic space of the ear, skeletal muscle cells, heart muscle cells, smooth muscle cells, myoepithelial cells, red blood cells, megakaryocytes, monocytes, connective tissue macrophages, Langerhan's cells, osteoclasts, dendritic cells, microglial cells, neutrophils, eosinophils, basophils, mast cells, plasma cells, helper T cells, suppressor T cells, killer T cells, immunoglobulin M, immunoglobulin G, immunoglobulin A, immunoglobulin E, killer cells, rod cells, cone cells, inner hair cells of the organ of Corti, outer hair cells of the organ of Corti, type I hair cells of the vestibular apparatus of the ear, type II cells of the vestibular apparatus of the ear, type II taste bud cells, olfactory neurons, basal cells of olfactory epithelium, type I carotid body cells, type II carotid body cells, Merkel cells, primary sensory neurons specialized for touch, primary sensory neurons specialized for temperature, primary neurons specialized for pain, proprioceptive primary sensory neurons, cholinergic neurons of the autonomic nervous system, adrenergic neurons of the autonomic nervous system, peptidergic neurons of the autonomic nervous system, inner pillar cells of the organ of Corti, outer pillar cells of the organ of Corti, inner phalangeal cells of the organ of Corti, outer phalangeal cells of the organ of Corti, border cells, Hensen cells, supporting cells of the vestibular apparatus, supporting cells of the taste bud, supporting cells of olfactory epithelium, Schwann cells, satellite cells, enteric glial cells, neurons of the central nervous system, astrocytes of the central nervous system, oligodendrocytes of the central nervous system, anterior lens epithelial cells, lens fiber cells, melanocytes, retinal pigmented epithelial cells, iris pigment epithelial cells, oogonium, oocytes, spermatocytes,

spermatogonium, ovarian follicle cells, Sertoli cells, and thymus epithelial cells, or combinations thereof.

In some embodiments, the cells may be enucleated cells, for example, red blood cells. In other embodiments, the cells may be nucleated cells. In other embodiments, the cells may be plant cells. In yet other embodiments, the cells may comprise a tissue, a plant, or an organ, in whole or in part.

In some embodiments, cells may be cooled at a rate to a temperature from about -30 to about -80°C or less. In some embodiments, cells may be frozen in liquid nitrogen, for example, at about -196°C. In some embodiments, the cells may be stabilized at higher temperatures such as a room temperature (e.g, about 20-22°C) and up to about 30°C. In some embodiments, cells may be subjected to a temperature from about -30°C to about 20°C (e.g, about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein), e.g, cells may be cooled at rate to a temperature from about -30°C to about 20°C (e.g, about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein). For example, in some embodiments, cells may be subjected to a temperature between -196°C about 30°C, between -80°C and 22 °C, between 0°C and 30°C, or between - 30°C and 20°C.

The term“tissue” as used herein, refers to a collection of single or multiple cell types. Non-limiting examples include connective tissue, muscle tissue ( e.g ., visceral (smooth) muscle tissue, skeletal muscle tissue, or cardiac muscle tissue), neural tissue generated in accordance with the methods described herein can comprise central nervous system tissue (e.g., brain tissue or spinal cord tissue) or peripheral nervous system tissue (e.g, cranial nerves and spinal nerves) and epithelial tissue (e.g, endothelium). “Plant tissue” as used herein means a group of plant cells organized into a structural and functional unit. Any tissue of a plant in planta or in culture is included. This term includes, but is not limited to, whole plants, plant organs, plant seeds, tissue culture and any groups of plant cells organized into structural and/or functional units. The use of this term in conjunction with, or in the absence of, any specific type of plant tissue as listed above or otherwise embraced by this definition is not intended to be exclusive of any other type of plant tissue.

In some embodiments, tissues may be cooled at a rate to a temperature from about -30 to about -80°C or less. In some embodiments, tissues may be frozen in liquid nitrogen, for example, at about -196°C. In some embodiments, the tissues may be stabilized at higher temperatures such as a room temperature ( e.g ., about 20-22°C) and up to about 30°C. In some embodiments, tissues may be subjected to a temperature from about -30°C to about 20°C (e.g., about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein), e.g, tissue may be cooled at rate to a temperature from about -30°C to about 20°C (e.g, about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein). For example, in some embodiments, tissues may be subjected to a temperature between -196°C about 30°C, between -80°C and 22 °C, between 0°C and 30°C, or between - 30°C and 20°C.

The term“organ” as used herein refers to a collection of tissues associated with any of the known mammalian organ systems, i.e., the digestive system, circulatory system, endocrine system, excretory system, immune system, integumentary system, muscular system, nervous system, reproductive system, respiratory system, and/or skeletal system. Exemplary organs that can be used in accordance with the methods described herein include, without limitation, lungs, liver, heart, brain, kidney, skin, bone, stomach, pancreas, bladder, gall bladder, small intestine, large intestine, prostate, testes, ovaries, spinal cord, pharynx, larynx, trachea, bronchi, diaphragm, ureter, urethra, esophagus, colon, thymus, and spleen.

In some embodiments, organs may be cooled at a rate to a temperature from about -30 to about -80°C or less. In some embodiments, organs may be subject to hypothermic temperatures. In some embodiments, organs may be frozen in liquid nitrogen, for example, at about -196°C. In some embodiments, the organs may be stabilized at higher temperatures such as a room temperature (e.g, about 20-22°C) and up to about 30°C. In some

embodiments, organs may be subjected to a temperature from about -30°C to about 20°C (e.g, about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein), e.g, organs may be cooled at rate to a temperature from about -30°C to about 20°C ( e.g ., about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein). For example, in some embodiments, organs may be subjected to a temperature between -196°C about 30°C, between -80°C and 22 °C, between 0°C and 30°C, or between -30°C and 20°C.

It is noted that a plant can include a“plant part” including, but not limited to, embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, stalks, roots, root tips, anthers, and / or plant cells including plant cells that are intact in plants and/or parts of plants, plant protoplasts, plant tissues, plant cell tissue cultures, plant calli, plant clumps, and the like.

Exemplary plants include, but are not limited to corn (Zea mays), canola (Brassica napus, Brassica rapa ssp.), alfalfa (Medicago saliva), rice (Oryza sativa, including without limitation Indica and/or Japonica varieties), rape (Brassica napus), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), sunflower (Helianthus annus), wheat

(Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tobacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihot esculenta), coffee (Cofea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), apple (Malus pumila), blackberry (Rubus), strawberry (Fragaria), walnut (Juglans regia), grape (Vitis vinifera), apricot (Prunus armeniaca), cherry (Prunus), peach (Prunus persica), plum (Prunus domestica), pear (Pyrus communis), watermelon (Citrullus vulgaris) duckweed (Lemna), oats (. Avena sativa ), barley ( Hordium vulgare ), vegetables, ornamentals, conifers, and turfgrasses (e.g, for ornamental, recreational or forage purposes), and biomass grasses (e.g, switchgrass and miscanthus).

Vegetables include solanaceous species (e.g, tomatoes; Lycopersicon esculentum), lettuce (e.g, Lactuea sativa), carrots (Caucus carota), cauliflower (Brassica oleracea), celery (apium graveolens), eggplant (Solanum melongena), asparagus (Asparagus officinalis), ochra (Abelmoschus esculentus), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), members of the genus Cucurbita such as Hubbard squash (C. Hubbard), Butternut squash (C. moschata), Zucchini (C. pepo), Crookneck squash (C.

crookneck), C. argyrosperma , C. argyrosperma ssp sororia, C. digitata, C. ecuadorensis, C. foetidissima, C. lundelliana, and C. martinezii, and members of the genus Cucumis such as cucumber (Cucumis sativus), cantaloupe (C. cantalupensis), and musk melon (C. melo).

Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (dianthus caryophyllus), poinsettia (Euphorbia pulcherima), and chrysanthemum.

Turf grasses include but are not limited to zoysia grasses, bent grasses, fescue grasses, blue grasses, St. Augustine grasses, Bermuda grasses, buffalo grasses, rye grasses, and orchard grasses.

Also included are plants that serve primarily as laboratory models, e.g., Arabidopsis.

“Reduce,”“reducing,”“reduction,” and grammatical variants thereof, as used herein, refer to a decreased effect relative to what would occur in the absence of the methods of the present invention. In particular, such effects according to the present invention include a decrease in ice crystal formation, a decrease in contamination (e.g, bacterial contamination) a decrease in cell death, and/or a decrease in function (e.g, biological function).

In some embodiments, plants may be cooled at a rate to a temperature between -30 and -80°C or less. In some embodiments, plants may be subject to hypothermic temperatures. In some embodiments, organs may be frozen in liquid nitrogen, for example, at -196°C. In some embodiments, the plants may be stabilized at higher temperatures such as a room temperature (e.g, about 20-22°C) and up to 30°C. In some embodiments, plants may be subjected to a temperature between -30°C and 20°C (e.g, about -30, -29, -28, -27, -26, -25, - 20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein), e.g, plants may be cooled at a rate to a temperature between -30°C and 20°C (e.g, about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein). For example, in some embodiments, plants may be subjected to a temperature between -196°C about 30°C, between -80°C and 22 °C, between 0°C and 30°C, or between -30°C and 20°C.

The term“biomaterial” and/or“inanimate biomaterial” as used herein may refer to any material in any form (e.g, whole (e.g, wood, e.g, a wooden implement) and/or otherwise processed (e.g, paper product; composite)) originally sourced from an organic source (e.g, biological source, e.g, living material, e.g, a plant), including, but not limited to, wood, cellulose, cartilage, and/or gelatin. The term“article of manufacture comprising inanimate biomaterial” as used herein, refers to inanimate objects at least partially generated from and/or otherwise comprising organic material (i.e., inanimate biomaterial). Non-limiting examples of articles of manufacture comprising inanimate biomaterial may include, e.g, paper and/or wooden implements. In some embodiments, inanimate biomaterial and/or an article of manufacture comprising inanimate biomaterial may comprise degraded inanimate biomaterial such as, but not limited to, waterlogged wood and/or archeological wood.

Without wishing to be bound to theory, it is believed that preservation of sugars may be able to preserve waterlogged or otherwise degraded wood by filling in the pockets (for example, by crystalizing in the pockets) created in the wood structure following degradation of cellulose.

In some embodiments, inanimate biomaterial may be cooled at a rate to a temperature from about -30°C to about -80°C or less. In some embodiments, inanimate biomaterial may be subject to hypothermic temperatures. In some embodiments, inanimate biomaterial may be frozen in liquid nitrogen, for example, at about -196°C. In some embodiments, inanimate biomaterial may be stabilized at higher temperatures such as a room temperature (e.g, about 20-22°C) and up to about 50°C. In some embodiments, inanimate biomaterial may be subjected to a temperature from about -80°C to about 50°C (e.g, about -80, -75, -70, -65, -60,

-55, -50, -45, -40, -35, -30, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 20, 25,

30, 45, or 50°C or any value or range therein), e.g, inanimate biomaterial may be cooled at a rate to a temperature from about -80°C to about 50°C (e.g, about -80, -75, -70, -65, -60, -55,

-50, -45, -40, -35, -30, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 45, or 50°C or any value or range therein). For example, in some embodiments, inanimate biomaterial may be subjected to a temperature from about -80 °C to about 45°C, about -65°C to about 35°C, about -45°C to about 30°C, or -30°C to about 20°C, e.g., inanimate

biomaterial may be cooled at a rate to a temperature from about -80 °C to about 45°C, about - 65°C to about 35°C, about -45°C to about 30°C, or -30°C to about 20°C.

In some embodiments, prior to preservation an inanimate biomaterial may be treated according to methods known in the art. For example, wood may be dried and/or seasoned by being heated to a particular temperature range to reduce moisture prior to preservation, such as, but not limited to, freeze-dried, air-dried while heated at ambient temperature or higher (e.g, heated to a temperature of about 20 °C to about 60 °C), and/or kiln-dried at a temperature of about 60 °C or higher (e.g, about 60 °C to about 120 °C).

“Suitable carrier for preservation” refers to a carrier that does not cause significant irritation to a cell, tissue, organ, or inanimate biomaterial and does not abrogate the biological activity and/or properties of the administered compound thereby facilitating preservation. “Suitable carrier for cryopreservation” refers to a carrier that does not cause significant irritation to a cell, tissue or organ and does not abrogate the biological activity and/or properties of the administered compound thereby facilitating cryopreservation and/or successful thawing.

“Kit” as used herein refers to an assembly of components. The assembly of components can be a partial or complete assembly. Instructions for use of the kit or use of various components of the kit are optionally included.

Embodiments of the present invention provide methods and compositions useful for preservation of cells, tissues, organs, and/or inanimate biomaterial with such methods and compositions useful for the treatment of diseases and disorders in a subject in need thereof in addition to the preservation of plants.

Compounds suitable for use according to the present invention include sucrose, sucralose, trehalose, and maltose. In some embodiments, the compound has the following structure for sucrose (A), sucralose (B), trehalose (C), and/or maltose (D):

In some embodiments, the compound is I ,6-Dichloro-1 ,6-dideoxy-|3-D- fructofuranosyl-4-c!iloro-4~deoxy-a-D-galactopyranoside; (2R,3R,4R,5R,6R)-2- [(2/^3,V,4,V,5,V)-2,5-Bis(chloromethyl)-3,4-dihydrox_\Oxolan-2-yl]oxy-5-chloro-6- (hydroxymethyl)oxane-3,4-diol; l',4,6'-trichlorogalactosucrose; trichlorosucrose; E955; 4, r,6'-Trichloro-4, r,6'-trideoxygalactosucrose or TGS.

In some embodiments the sucrose, sucralose, trehalose and/or maltose is present in a concentration in range from about 0.01% w/v to about 100% w/v, 10% w/v/ to 100% w/v, 30% w/v to 75% w/v, 40% w/v to 100% w/v, 15% w/v to 65%w/v, 0.01% w/v to 30% w/v, 0.01% w/v to 20% w/v, 0.01% w/v to 10% w/v, 0.01% w/v to 1% w/v, 0.1% w/v to 1% w/v, 0.1% w/v to 1.5% w/v, 0.1% w/v to 2% w/v, or 0.1% w/v to 2.5% w/v, and including any numerical values included within the specified range limits. For example, sucrose, sucralose, trehalose and/or maltose concentrations may be about 0.125% w/v, 0.250% w/v, 0.375% w/v, 0.500% w/v, 0.625% w/v, 1.000% w/v, 1.250% w/v, 1.500% w/v, 1.750% w/v, 2.000% w/v, 2.25% w/v, 2.500% w/v, 25.000% w/v, 40.175% w/v, or 60.575% w/v.

In particular embodiments, sucrose may be used alone, sucralose may be used alone, trehalose may be used alone, maltose may be used alone, or sucrose, sucralose, trehalose and/or maltose may be used in combination in various ratios as determined by one of ordinary skill in the art to obtain the desired result.

In some embodiments, sucrose, sucralose, trehalose and/or maltose may dehydrate the cell membrane thereby reducing or eliminating ice crystal formation and/or growth.

Embodiments of the present invention include methods of preparing a cell, tissue, organ, plant, or inanimate biomaterial for preservation, the method comprising, consisting essentially of, or consisting of contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising, consisting essentially of, consisting of sucrose, sucralose, trehalose and/or maltose. Cells, tissues, organs, plants, and inanimate biomaterial used in accordance with this invention are described above. In some embodiments, the cells are red blood cells.

In some embodiments, the present invention provides methods for the preservation of a cell, tissue, plant or organ, comprising, consisting essentially of, or consisting of (a) contacting the cell, tissue, plant or organ with a composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose; and (b) subjecting the cell, tissue, plant or organ to a temperature from about -30°C to about 20°C (e.g, about - 30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein). In some embodiments, the cell, tissue, organ or plant are contacted with the composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose before step (b), during step (b), and/or after step (b).

In some embodiments, the present invention provides methods for the preservation of inanimate biomaterial, comprising, consisting essentially of, or consisting of (a) contacting the inanimate biomaterial with a composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose; and (b) subjecting the inanimate biomaterial to a cryogenic temperature ( e.g ., frozen in liquid nitrogen, e.g., at about -196°C) and/or a temperature from about -80°C to about 50°C (e.g, hypothermic temperatures up to room temperature or warmer, e.g, about -80, -75, -70, -65, -60, -55, -50, -45, -40, -35, -30, - 25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 45, or 50°C or any value or range therein). For example, in some embodiments, the method may comprise (b) subjecting the inanimate biomaterial to a temperature from about -80°C to about 30°C, a temperature from about -60°C to about 40°C, a temperature from about -30°C to about 20°C, or a temperature from about -30°C to about 10°C. In some embodiments, the inanimate biomaterial is contacted with the composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose before step (b), during step (b), and/or after step (b).

Embodiments of the present invention also provide methods of reducing ice crystal formation in a cell, tissue, organ or plant, the method comprising, consisting essentially of, or consisting of contacting the cell, tissue, organ or plant with a composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a cryogenic temperature. In some embodiments, the methods provide vitrification. In addition to reducing ice crystal formation, the methods provide greater success rates of obtaining suitable thawed cells, tissues, organs or plant for use after cryopreservation, i.e., improved recovery upon thawing.

In accordance with the present invention, further embodiments provide methods of reducing cell death during preservation of a cell, tissue, organ or plant, the method comprising, consisting essentially of, or consisting of contacting the cell, tissue, organ or plant with a composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C. In some embodiments, the cell, tissue, organ or plant are contacted with the composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose before, during and/or after being subjected to the temperature range. Thus, in addition to reducing cell death during preservation, the methods provide greater success rates of maintaining suitable cells, tissues, organs or plants for use after storage, i.e., improved retention upon storage.

In some embodiments, the present invention provides a method of preserving cell function during preservation of a cell, tissue, organ or plant, the method comprising contacting the cell, tissue, organ or plant with a composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature ( e.g ., a freezing temperature from about -30°C to about -80°C, a cryogenic temperature, a temperature from about -30°C to about 20°C, and/or room temperature), wherein the cell function is reduced by less than about 50% (e.g., by less than about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50%, or any value or range therein) as compared to a control. In some embodiments, a control may be, e.g, not contacting the cell, tissue, organ or plant with a composition comprising sucrose, sucralose, trehalose, and/or maltose prior to subjecting the cell, tissue, organ or plant to a temperature (e.g, a freezing temperature from about -30°C to about -80°C, a cryogenic temperature, a temperature from about -30°C to about 20°C, and/or room temperature), and/or e.g, an unpreserved cell, tissue, organ or plant. For example, in some embodiments, the method of the present invention may comprise wherein the cell function is reduced by less than about 5% to about 15%, or about 2% to about 50%, or about 0.5% to about 40%, etc. In some embodiments, the cell may be red blood cells, and the function may be carrying oxygen.

In some embodiments, the present invention provides a method of reducing contamination during preservation of a cell, tissue, organ, plant, or inanimate biomaterial, the method comprising contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature (e.g, from about -80°C to about 50°C, e.g, from about -30°C to about 20°C), wherein the contamination is reduced by more than about 50% (e.g, by more than about 50, 55, 60, 65,

70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 100, 150, or 200% or more) as compared to a control (e.g, not contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising sucrose, sucralose, trehalose, and/or maltose prior to subjecting the cell, tissue, organ, plant, or inanimate biomaterial to a particular temperature (e.g, from about -80°C to about 50°C , e.g, from about -30°C to about 20°C), and/or e.g, an unpreserved cell, tissue, organ, plant, or inanimate biomaterial). In some embodiments, the contamination may be bacterial and/or fungal (e.g, mold) contamination.

In some embodiments of the present invention, methods of improved preservation of a cell, tissue, organ, or inanimate biomaterial as compared to preservation of a cell, tissue, organ, plant, or inanimate biomaterial with a saline-adenine-glucose-mannitol preservative or derivative thereof (e.g, SAGM, AS1, and/or AS3), are provided, the method comprising, consisting essentially of, consisting of contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising, consisting essentially of, consisting of sucrose, sucralose, trehalose, and/or maltose. In some embodiments of the present invention, methods for lyopreservation of liposomes are provided, comprising (a) contacting a liposome comprising a drug with a composition comprising sucrose, sucralose, trehalose and/or maltose; and (b) subjecting the liposome to a temperature from about -30°C to about 20°C (e.g, about -30, -29, -28, -27, -26, -25, -20, -15, -10, -5, -4, -3. -2, -1, 0, 1, 2, 3, 4, 5, 10, 15, 16, 17, 18, 19, or 20°C or any value or range therein).

Embodiments of the present invention further provide a preservation medium comprising, consisting essentially of, or consisting of sucrose, sucralose, trehalose and/or maltose in a carrier suitable for preservation and/or cryopreservation.

Embodiments of the present invention also provide preserved and/or cryopreserved preparations of cells, tissues, organs, or inanimate biomaterial prepared by the methods recited herein.

In particular embodiments, the present invention provides compositions such as pharmaceutical compositions comprising, consisting essentially of, or consisting of the cells subjected to the methods described herein; transfusable preparations comprising, consisting essentially of, or consisting of the preserved and/or cryopreserved cells described herein; transplantable preparations comprising, consisting essentially of, or consisting of the tissues, organs, or inanimate biomaterial subjected to the methods described herein.

Embodiments of the present invention further provide kits comprising, consisting essentially of, consisting of one or more containers comprising, consisting essentially of, or consisting of the cells subjected to the methods described herein and/or components for the preservation and/or cryopreservation of cells, tissues, organs, plants, and/or inanimate biomaterial.

Some embodiments of the present invention are directed to use in subjects such as those described above. Additionally, subjects further include, but are not limited to, those in need of a cell transfusion and/or tissue and/or organ transplant.

When administered or used in a pharmaceutical manner, the preserved cells, tissues, organs, and/or inanimate biomaterial may be maintained and/or thawed and suitable for the intended use. They may be combined with a physiologically acceptable carrier. In some embodiments, the physiologically acceptable carrier can include, but is not limited to, sterile water, saline, glucose, dextrose, stabilizers (e.g, sugars and amino acids), preservatives, wetting agents, emulsifying agents, and pH buffering agents. Suitable carriers for pharmaceutical compositions are described in Remington’s Pharmaceutical Sciences by E.

W. Martin. Suitable ranges of temperatures for the methods recited herein are discussed above. Cooling rates are generally fast, for example, up to 100°C/min. In terms of storage, once stabilized, storage time is generally directly related to the sugar stability/shelf life.

Having described the present invention, the same will be explained in greater detail in the following examples, which are included herein for illustration purposes only, and which are not intended to be limiting to the invention.

EXAMPLES

EXAMPLE 1: Refrigeration of non-diluted blood

Blood washing was performed on defibrillated sheep blood (30 mL) purchased from Hemostat Laboratories. A sample of blood was washed by centrifugation at 500 RCF at 22°C for 6 minutes and removing the supernatant and replacing it with 150 mM NaCl. The washing process was repeated until the supernatant was clear.

Aliquots of washed blood were treated with stock sugar solution of either sucralose or sucrose and buffer to obtain the required sugar concentration (0.0%-1.0% w/v) while maintaining equal sample volumes. The sugar was added in 20 pL aliquots followed by gentle mixing after each addition to minimize osmotic shock. After the addition of sugar, samples were incubated for 1 hour at room temperature and then transferred to the refrigerator. Aliquots of each sample were removed at varying time intervals and analyzed for percent hemolysis using previously described methods by measuring the absorbance of the supernatant in a 96 well plate reader at 540 nm. Control data for 0% and 100% hemolysis were obtained using buffer and blood treated with triton, respectively. The data obtained are shown in FIGS. 1A-1B and 2A-2B.

EXAMPLE 2

FIGS. 3 and 4 show the results of long term studies where blood was stored in a refrigerator after treatment with various protective solutions.

Prior to treatment, packed red sheep blood was washed with sodium chloride until the buffy coat was clear and each time the top layer was removed and replaced with PBS buffer. The blood was then dispensed into Eppendorf tubes and treated with an appropriate solution to achieve the final concentration of AS1 , sucralose (% w/v), or a combination of AS1 combined with sucralose (%w/v). The final volume of each sample was equal and samples were stored undisturbed in the refrigerator until required for analysis. Hemolysis was determined by measuring free hemoglobin using UV absorption spectrophotometry in a 96 well plate, versus buffer for negative control and samples treated with 1% Triton-X for maximum hemolysis.

FIG. 3 displays hemolysis of samples treated with varying quantities of sucralose. The sucralose concentrations are given as % w/v. It can be seen that samples treated with 1% sucralose have considerably lower hemolysis than untreated samples after 70 days storage. It also appears that sucralose may offer protection in a dose dependent manner.

FIG. 4 shows relative absorbance (a surrogate for percent hemolysis) for samples treated with a combination of AS1 and sucralose, compared to AS1 alone and untreated blood. Data seem to indicate improved protection when small quantities of sucralose are added to AS 1. The data appear to indicate that sucralose does not interfere with the protective ability of AS 1.

The foregoing is illustrative of the present invention and is not to be construed as limited to the specific embodiments disclosed, and that modifications to the disclosed embodiments, as well as other embodiments, are intended to be included within the scope of the appended claims. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

That which is claimed is:

1. A method for the preservation of a cell, tissue, organ or plant, comprising:

(a) contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose; and

(b) subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C.

2. A method of reducing cell death during preservation of a cell, tissue, organ or plant, comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C.

3. A method of preserving cell function ( e.g ., carrying oxygen) during preservation of a cell, tissue, organ or plant, comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C, wherein the cell function is reduced by less than about 50% (e.g., by less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50%, etc.) as compared to a control (e.g, not contacting the cell, tissue, organ or plant with a composition comprising sucrose, sucralose, trehalose, and/or maltose prior to subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C, and/or e.g, an unpreserved cell, tissue, organ or plant).

4. A method of reducing bacterial contamination during preservation of a cell, tissue, organ or plant, comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C, wherein the bacterial contamination is reduced by more than about 50% (e.g, by more than about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 100, 150, 200% or more) as compared to a control (e.g, not contacting the cell, tissue, organ or plant with a composition comprising sucrose, sucralose, trehalose, and/or maltose prior to subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C, and/or e.g, an unpreserved cell, tissue, organ or plant).

5. The method of claim 1, wherein the method comprises improvement in the preservation of a cell, tissue, organ or plant compared to preservation of a cell, tissue, organ or plant with a saline-adenine-glucose-mannitol preservative or derivative thereof (e.g, SAGM, AS1, AS3), comprising contacting the cell, tissue, organ or plant with a composition comprising about 0.01% to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the cell, tissue, organ or plant to a temperature from about -30°C to about 20°C.

6. The method of any one of claims 1-5, wherein the cell is a red blood cell.

7. The method of any one of claims 1-5, wherein the plant is a tomato plant.

8. A method for the preservation of inanimate biomaterial, comprising:

(a) contacting the inanimate biomaterial with a composition comprising about 0.01 % to about 2.5% w/v sucrose, sucralose, trehalose, and/or maltose; and

(b) subjecting the inanimate biomaterial to a temperature from about -80°C to about

50°C.

9. A method of reducing bacterial contamination during preservation of an inanimate biomaterial, comprising contacting the inanimate biomaterial with a composition comprising 0.01% to 2.5% w/v sucrose, sucralose, trehalose, and/or maltose and subjecting the inanimate biomaterial to a temperature from about -80°C to about 50°C, wherein the bacterial contamination is reduced by more than about 50% ( e.g ., by more than about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 100, 150, 200% or more) as compared to a control (e.g., not contacting the inanimate biomaterial with a composition comprising sucrose, sucralose, trehalose, and/or maltose prior to subjecting the inanimate biomaterial to a temperature from about -80°C to about 50°C, and/or e.g, an unpreserved inanimate biomaterial).

10. The method of any one of claims 1-9, wherein the method comprises contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising sucrose.

11. The method of any one of claims 1-9, wherein the method comprises contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising sucralose.

12. The method of any one of claims 1-9, wherein the method comprises contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising trehalose.

13. The method of any one of claims 1-9, wherein the method comprises contacting the cell, tissue, organ, plant, or inanimate biomaterial with a composition comprising maltose.

14. A preserved preparation of cells, tissues, organs or plants or inanimate biomaterial prepared by the method of any one of claims 1-5 or claim 8 or 9.

15. A pharmaceutical composition comprising a cell subjected to the method of any one of claims 1-5.

16. A method for the lyopreservation of liposomes, comprising:

(a) contacting a liposome comprising a drug with a composition comprising sucrose, sucralose, trehalose and/or maltose; and

(b) subjecting the liposome to a temperature from about -30°C to about 20°C.

17. The method of claim 16, wherein the drug is selected from the group consisting of a cardiovascular, analgesic, antibiotic, antiviral, antimicrobial,

chemotherapeutic, gastrointestinal, central nervous system, antidepressant, respiratory, anti inflammatory, hormone, contraceptive, vitamin, antihistamine, anti-allergy, insulin, antidiabetic and antimalarial drug.

18. The method of claim 16, wherein the drug is doxorubicin, daunorubicin, amphotericin, vincristine, verteporfm, cytarabine, morphine amikacin, cisplatin and/or paclitaxel.