Abstract: The invention can be summarized as follows. The present invention provides cell culture compositions and cell cryopreservation compositions for culturing and cryopreserving enteric system (ENS) neurons and central nervous system (CNS) neurons. The present invention also provides a method of culturing and cryopreserving ENS and CNS cells. The compositions of the present invention comprise nutrient medium, serum, glutamine, glucose an antibiotic and one or more of the following components: one or more mitotic inhibitors, one or more antioxidants, and a cryopreservative.

Abstract: A sample support (100, 101, 102, . . . ), in particular for cryopreservation of biological samples, is described and includes: a holder part (10) with a sample receiving chamber (11), a retaining device (12, 13) on which a sample can be disposed, and support connecting elements (14, 15) which are directed at opposite holder part ends (16, 17) and are formed to fit together so that a plurality of holder parts (16) can be put together. A sample store (200), which includes an assemblage of a large number of such sample supports, and a method for the cryopreservation of biological samples, are also described.

Abstract: A cryoprotective composition which comprises nanostructures, liquid and at least one cryoprotective agent is provided.

Abstract: A cryopreservation storage device (100), in particular for cryopreservation of biological samples, comprises a plurality of multi sample modules (20) being adapted for accommodating the biological samples and sample memories, a module control device (30) controlling an access to sample memories accommodated by the multi sample modules (20), and a data interface (41) for accessing to the module control device (30), wherein the module control device (30) includes a data management processor (31), which can be controlled via the data interface (41). Furthermore, a cryopreservation apparatus including at least one cryopreservation storage device (100) and a method for cryopreservation of biological samples are described.

Abstract: The present invention relates to a method for improving viability and/or stress tolerance of viable biological material and using the said material comprising applying hydrostatic pressure to said biological material; keeping the said viable biological material at the hydrostatic pressure for a predetermined time period; releasing the hydrostatic pressure; and using the said material for any desired purpose in accordance with any useful protocol. The usage of the said biological material incorporates any techniques, protocols that are applicable in the field of assisted reproductive techniques, biotechnical and/or biotechnological manipulations.

Abstract: A double-structured tissue implant and a method for preparation and use thereof for implantation into tissue defects. The double-structured tissue implant comprising a primary scaffold and a secondary scaffold consisting of a soluble collagen solution in combination with a non-ionic surfactant generated and positioned within the primary scaffold. A stand alone secondary scaffold implant or unit. A process for preparation of the double-structured implant or the stand alone secondary scaffold comprising lyophilization and dehydrothermal treatment.

Abstract: The present invention provides a paraffin-penetration treatment method for a tissue preparation, which is capable of enabling sufficient penetration of paraffin even into a tissue preparation, into which paraffin can hardly penetrate by a conventional paraffin-penetration treatment method. When the penetration of the paraffin into the tissue preparation is performed by immersing the tissue preparation in molten paraffin after a dehydrating and a degreasing treatments, the method comprises the steps of: immersing the tissue preparation in the molten paraffin so as to allow the paraffin to penetrate into the tissue preparation; cooling the tissue preparation so as to solidify the paraffin having penetrated thereinto; and immersing the tissue preparation in the molten paraffin again so as to melt the paraffin having penetrated thereinto, and the above described solidification treatment and melting treatment are conducted at least once for one tissue preparation.

Abstract: [Problem] To provide a preservative solution for cells, tissues and organs using a rare sugar and a preservation method using the solution. [Means for Resolution] A preservative solution for cryopreservation of animal or human organs and animal or plant tissues or cells, the solution containing a rare sugar D-allose. A preservative solution for cryopreservation of the human kidney, cells derived from animals or humans, or human or animal sperm and/or ovum and/or fermented ovum in which the cryopreservation is conducted at from ?5 to 20° C. A preservative solution for cryopreservation of cells derived from animals or humans, or human or animal sperm and/or ovum and/or fermented ovum in which the cryopreservation is conducted at a temperature at which to start freezing to ?196° C.

Abstract: The subject invention pertains to compositions and methods for promoting repair of damaged nerve tissue using nerve grafts and preparation of nerve grafts. The compositions and methods of the subject invention can be employed to restore the continuity of nerve interrupted by disease, traumatic events or surgical procedures. Compositions of the subject invention comprise one or more chondroitin sulfate proteoglycan (CSPG)-degrading enzymes that promote axonal penetration into damaged nerve tissue and nerve graft. The invention also concerns methods for promoting repair of damaged nerve tissue using the present compositions and nerve tissue treated according to such methods. The invention also includes storage solutions for nerve tissue.

Abstract: Cryopreserved cultures of post-mitotic neuronal or neural-like cells are provided having a viability after thaw of greater than 10%, typically greater than 50%. Once thawed, the cells are capable of further differentiation. In one embodiment, less than 20% of the cryopreserved cells are self-proliferating cells. The cells can be provided in a kit including a container of the cryopreserved neuronal or neural-like cells, optionally including additional cell culture reagents and materials. Method for preparing the cryopreserved neuronal or neural-like cells derived from embryonic stem cells, preferably human embryonic stem cells, are also provided.

Abstract: A cryopreservation medium comprising a protein-containing plant extract is disclosed. Methods, compositions, uses and kits for cryopreservation of biological material, such as a molecule, organelle, cell, embryo, tissue or organ, are also disclosed.

Abstract: The invention relates to methods of processing amniotic membrane to generate a substantially ‘growth factor free’ membrane (GFF-membrane), and to methods of processing GFF-membrane to generate membrane enriched with specific and quantified levels of growth factors or other desirable membrane enriching molecules or compounds (E-membrane). The invention extends to GFF-membrane per se and E-membrane enriched with specific and quantified membrane-enriching compounds per se. The method also includes first and second medical uses of GFF-membrane and E-membrane. The method also extends to clinical uses of the amniotic membrane spongy layer or components thereof.

Abstract: Methods and compositions for the preservation of cells using glucosaminoglycans and derivatives thereof.

Abstract: The present invention is directed to compositions of fetal hematopoietic stem and progenitor cells and stromal (mesencymal) cells derived from second trimester (16-20 weeks gestation) miscarriages, that are isolated, processed and cryopreserved, and the therapeutic uses of such stem and progenitor cells upon application. Such compositions may be useful for hematopoietic reconstitution and/or replacement in patients with various diseases and metabolic disorders. The invention also relates to methods for collecting, isolating, processing and cryopreserving of the fetal stem and progenitor cells of the invention. Importantly, the invention meets an urgent need in regenerative medicine while at the same time ensures utility of the product compositions because it utilizes fetal materials from second trimester (16-20 weeks of gestation) spontaneously lost pregnancies which are free of moral or ethical burdens.

Abstract: The present invention provides methods for providing cartilage-containing tissue for grafting, comprising providing excised cartilage-containing tissue; and treating said excised cartilage-containing and cryogenically preserving the treated cartilage-containing tissue under appropriate cryogenic preservation conditions so as to yield cryogenically preserved cartilage-containing tissue having at least 10% viable chondrocytes throughout the cartilage portion of the cartilage-containing tissue after preservation, as tested in a live/dead ratio assay. Treatment may comprise providing one or a plurality of incisions in said cartilage portion to a predetermined depth therein and/or introducing a cryoprotectant agent at least into said cartilage portion.

Abstract: A method for determining the effectiveness of a treatment for preeclampsia of a pregnant woman at risk for preeclampsia, the method comprising: (a) determining a first concentration of placental protein 13 (PP13) in a bodily substance of the woman obtained prior to the treatment; (b) determining a second concentration of PP13 in a bodily substance of the woman obtained after initiation of the treatment; and (c) comparing the first and second concentrations to a corresponding normal level of PP13 and, based on the comparison, determining the effectiveness of the treatment. Diagnostic kits for practicing the method are also disclosed.

Abstract: A method of forming and preserving a bioremodelable, biopolymer scaffold material by subjecting animal tissue, particularly fetal or neo-natal tissue, to chemical and mechanical processing. The process includes, but is not limited to, harvesting the tissue, optionally extracting growth and differentiation factors from the tissue, inactivating infective agents of the tissue, mechanically expressing undesirable components from the tissue, delipidizing the tissue, washing the tissue, optionally drying the tissue, optionally cross-linking the tissue not necessarily in the order described. The resulting product, EBM, is characterized by its microbial, fungal, viral and prion inactivated state. EBM is strong, bioremodelable, drapable and does not undergo calcification. EBM supplants previous inventions because of its unique method of preparation and broad applicability in tissue reengineering.

Abstract: The present invention relates, in general, to tissue decellularization and, in particular to a method of treating tissues, for example, heart valves, tendons and ligaments, so as to render them acellular and thereby limit mineralization and/or immunoreactivity upon implementation in vivo.

Abstract: An egg freezing and storing instrument has an egg freezing and storing tube made of a liquid nitrogen-resistant material; and a metal cylindrical protection member for protecting the tube. The tube has a body part; and an egg-storing small-diameter part having an inner diameter of 0.1 mm to 0.5 mm. The tube can be heat-sealed at a front side of the small-diameter part and at a rear side of the body part. The cylindrical protection member has a tubular part for accommodating a front side of the small-diameter part of the tube; and a semi-tubular part for accommodating a portion of the small-diameter part not accommodated in the tubular part and a front portion of the body part.

Abstract: A method is disclosed for reliably stabilizing eukaryotic cells that express the P2X7 receptor channel, particularly mammalian and other vertebrate cells, including human cells, for example mammalian macrophages, or hematopoietic stem cells, in order to introduce otherwise membrane-impermeable compounds that are helpful for stabilizing the cells during drying, chilling, freezing, freeze-drying, or cryopreservation. The cells are exposed to extracellular ATP in concentration sufficient to open pores in the plasma membrane. One or more otherwise membrane-impermeable compounds that aid the survivorship of cells are then introduced, for example, trehalose, and after a brief time the pores are closed—for example, by adding divalent cations, or by diluting the extracellular solution. Once the trehalose or other stabilizing compound has been introduced, the cells may be stably preserved. By taking advantage of an endogenous mammalian receptor and ATP, no antigenic compounds need be introduced.

Abstract: Living cellular material may be preserved by incubating the cellular material in a culture medium containing at least one sugar, particularly for at least three hours, and then subjecting the cellular material to a preservation protocol, such as freezing, vitrification, freeze-drying and desiccation.

Abstract: The invention concerns an apparatus for preparing monolayers of cells. The apparatus comprises a container (6) for a cryosubstitution system and an insert (1) for the container, the insert (1) having a surface (1a) and a plurality of orifices (3). An SCS (2) together with a cellular monolayer (20) is insertable into each of the orifices (3). The SCS (2) is thus arranged perpendicular to the surface (1a) of the insert (1).

Abstract: A disclosure is made of various apparatus and methods for culturing and preserving cells and tissue in ways that minimize contamination potential, direct cells to reside in desired areas, allow uniform cell distribution during seeding, provide optimal growth conditions by controlling the amount of medium residing in proximity of cells, allow desired compounds and molecules to reside in proximity of the cells, allow co-culture, provide for efficient scale up, allow a desired shape of tissue to be created while retaining a closed system, and allow cryopreservation and reconstitution of cell and tissue while retaining a closed system. The apparatus and methods can be combined to prevent the need to remove the tissue from the enclosure at any point during the sterilization, seeding, culturing, cryopreservation, shipping, or restoration process. Also disclosed is an apparatus and method of pipette interface with a container in a manner that blocks contaminants from entering the container.

Abstract: This invention provides compositions and methods for cryoprotection of recombinant live cancer cells. Specifically, an improved cryoprotective medium is provided which includes a hydroxyethyl starch and/or derivative thereof alone or in combination with either DMSO or glycerol.

Abstract: A method for vitrification of a tissue or organ includes immersing the tissue or organ in increasing concentrations of cryoprotectant solution at a temperature greater than ?15° C. to a cryoprotectant concentration sufficient for vitrification; cooling the tissue or organ at an average rate of from 2.5–100° C. per minute to a temperature between ?80° C. and the glass transition temperature; and further cooling the tissue or organ at an average rate less than 30° C. per minute to a temperature below the glass transition temperature to vitrify the tissue or organ. After the vitrified tissue or organ has been stored, the tissue or organ may be removed from vitrification by warming the tissue or organ at an average rate of from 20–40° C. per minute to a temperature between ?80° C. and the glass transition temperature; further warming the tissue or organ at a rate greater than 80° C. per minute to a temperature above ?75° C.; and reducing the concentration of the cryoprotectant.

Abstract: This invention relates to formulations comprising biological samples preserved as dry glassy powders and hydrophobic carriers, the formulations being adapted for the long-term storage and delivery of the bioactive materials, in particular viral and bacterial vaccines, vectors and cells, at ambient or higher temperatures, and to methods for preparing these formulations.

Abstract: The present invention provides a composition for freezing or freeze-drying spermatozoa or sperm heads thereof, wherein the composition enables the spermatozoa or sperm heads thereof to maintain chromosome integrity at a temperature of about +4° C. to about ?200° C. Also provided is a container containing the composition. The present invention further provides a method for maintaining chromosome integrity of spermatozoa or sperm heads during freezing or freeze-drying. The present invention also provides methods for freezing or freeze-drying spermatozoa to obtain at least one spermatozoan capable of fertilizing an oocyte to produce a live offspring. Also provided are frozen or freeze-dried spermatozoa produced by these methods, and containers containing the frozen or freeze-dried spermatozoa. Finally, the present invention provides methods for producing a live mammalian offspring from an oocyte fertilized with a rehydrated freeze-dried (or thawed or freeze-thawed) spermatozoan.

Abstract: This invention relates to the sterilization and preservation of tissue for storage. More specifically, the invention relates to a method of using a solution to form a preserved tissue. The solution includes a radical scavenger at a concentration believed to reduce the damage to the tissue that could otherwise occur during sterilization with ionizing radiation.

Abstract: A preservation method for biological material having cell membranes includes microinjecting the cells with sugar; preparing the cells for storage; storing the biological material; and recovering the stored biological material from storage. The invention also features a method of culturing a cell in vitro using a hypertonic medium. Carbohydrate sugars such as trehalose, sucrose, fructose, dextran, and raffinose, may be used as bio-protective agents or in the culture medium.

Abstract: Methods of fixing and processing tissue and samples on a membrane by using ultrasound radiation as a part of the method are presented. Ultrasound of a frequency in the range of 0.1–50 MHz is used and the sample or tissue receives 0.1–200 W/cm2 of ultrasound intensity. The use of ultrasound allows much shorter times in the methods. Also presented are apparati comprising transducers of one or of multiple heads for producing the ultrasound radiation and further comprising a central processing unit and optionally comprising one or more sensors. Sensors can include those to measure and monitor ultrasound and temperature. This monitoring system allows one to achieve accurate and optimum tissue fixation and processing without overfixation and tissue damage. The system also allows the performance of antigen-antibody reactions or nucleic acid hybridizations to be completed in a very short time while being highly specific and with a very low or no background.

Abstract: The present invention relates to systems for the vitrification of a biological specimen. The systems employ a transfer instrument such as a loop, containing a base medium that includes a cryoprotectant. The biological specimen which has undergone vitrification may be stored for a period of time, and then thawed at a later date. The thawed biological specimen remains viable. Preferred biological specimens according to the present invention are developmental cells.

Abstract: A method of preserving, storing and transplanting mammalian donor organs. The method includes the cooling of refrigeration preservation, loading pre-freezer preservation, cryopreservation and washing solutions at least containing polyvinylpyrrolidone, a calcium channel blocker, a nucleoside, potassium chloride, polyethylene glycol, at least one amino acid, and a steroid to a temperature of 2° to 4° C. and/or of 0° to 2° C., harvesting a donor organ, perfusing it with one or more of the solution, immersing it in one or more of the solutions and storing it at a temperature above 0° C. or at a temperatures below 0° C. The cryopreservation solution also contains cryopreservative agents. Preserved organs may be transplanted directly from refrigeration storage or from freezer storage by cooling the washing refrigeration preservation solutions to 2° to 4° C., perfusing the organ with washing solution and then preservation solution, and transplanting it.

Abstract: In a method for cryopreserving fresh human oocytes, freezing and thawing solutions are used, which solutions include 1,2 propanediol (PROH) and sucrose at a concentration of at least 0.3M. The oocytes are exposed for 13–15 minutes to a solution including 1.5M PROH and 0.3M sucrose before starting the freezing process.

Abstract: This invention relates to sequential methods of cryopreserving bone marrow stromal cells that are transfected and used for gene therapy by transplantation. These methods include the following steps in various orders: obtaining the cells, expanding the cells in culture, transfecting the cells, and cryopreserving the cells. With these methods, populations of bone marrow stromal cells can be acquired that are large enough to be useful in a number of therapies. Further, these large populations can be stored for extended periods of time for immediate use when needed.

Abstract: A new method of cryopreservation based on the very fast cooling rates achieved by the direct contact of small droplets of vitrification solution containing biological sample with a very cold solid surface.

Abstract: A tissue of a multicellular organism is gradually dried during cultivation. After the tissue has been completely dehydrated, water is added to the tissue for its recovery. The tissue of the multicellular organism is submerged in an insect body fluid medium treated with heat, and dried for 48 hours or more.

Abstract: Methods and products relating the preservation of cultured mammalian epithelial or mesenchymal cells are provided. The methods involve the pre-treatment of epithelial cells with a solution containing a cryoprotectant amount of monosaccharides and/or disaccharides. The treated cells then are frozen, dried or freeze-dried in a minimum volume of solution and stored for later use. The invention avoids the use of materials that must be washed away from the preserved tissue prior to application of the tissue to a wound bed. The invention also permits the storage of epithelial tissue in a dry state.

Abstract: Compositions, methods, systems/devices and media are provided for maintaining a harvested organ in a functioning and viable state prior to implantation. The organ perfusion apparatus includes a preservation chamber for storing the organ during the preservation period. A perfusion circuit is provided having a first line for providing an oxygenated fluid to the organ, and a second line for carrying depleted fluid away from the organ. The perfusion apparatus also includes a device operably associated with the perfusion circuit for maintaining the organ at a substantially normothermic temperature.

Abstract: The present invention is directed to a cryoprotective system that comprises an aqueous solution containing polymeric nano- and micro-particles that exhibit a reversible temperature-dependent volume change. It is also directed to a method for providing cryoprotection to an organism or parts of an organism by pumping the cryoprotective system into the vasculature of the organism prior to exposing the organism to a lower, preferably below 0° C., temperature.

Abstract: Polyglycerol, lactose, and a combination of polyglycerol and lactose are effective at preserving cells, tissues, and organs from damage due to hypothermic, ischemic, or other metabolic impairment, and a mixture of polyglycerol plus lactose is particularly useful for the hypothermic storage of cells, tissues, and organs. The mixture of polyglycerol and lactose can be further improved by the addition of chondroitin sulfate, chlorpromazine, calcium, citrate, glutathione, adenine, glucose, magnesium, and a pH buffer.

Abstract: A physiological liquid medium is provided having the basic, synergistic components to allow its universal application in preserving cellular and functional integrity in vitro of different cell, tissue and organ types isolated from different mammalian species. The medium comprises an aqueous solution in sterile purified water of: i) a salt component comprising: a) from 100 to 150 mmoles/L of sodium ions, b) from 2.5 to 6.2 mmoles/L of potassium ions, c) from 1.0 to 2.5 mmoles/L of calcium ions, d) from 0.

Abstract: The invention relates to a two-step method for dehydrating biological tissues for producing preserved transplants. In a first step, the tissue is partially dehydrated with an organic, water-miscible solvent. In a second step, the tissue is dehydrated further by freeze drying.

Abstract: A biocompatible graft material and a method for making the same are disclosed. The method of making the graft material involves freezing and subsequently thawing a donated tissue sample in a bleach solution. The tissue is then washed in a detergent solution, treated with antimicrobial agents, and soaked in a hypertonic solution. The tissue is thereafter treated with sodium hydroxide and later hydrogen peroxide to yield the desired biocompatible graft material.

Abstract: The invention provides methods for the hypothermic preservation of cells, tissues and organs. In particular, the invention provides methods for the hypothermic preservation or storage of cells, tissues or organs in the vitreous state.

Abstract: A method of preserving functionality of an organ. The method includes removing a whole organ and associated vasculature, cryo-preserving the whole organ, allowing a period of time to elapse, thawing the whole organ including vasculature and introducing the whole organ into a recipient so that the organ is supplied with blood by vasculature belonging to the recipient. Further disclosed is a method of preserving fertility of a patient undergoing a treatment expected to cause sterility. The method include removing a whole gonadal organ from the patient, cryo-preserving the whole gonadal organ, conducting the treatment and waiting for an effect thereof to subside, thawing the whole gonadal organ and introducing the whole gonadal organ where it is supplied with blood by the vasculature system of the patient.

Abstract: A preservation method for biological material having cell membranes includes reversibly porating the cell membranes; loading a bio-protective agent having bio-preservation properties to a predetermined intracellular concentration; preparing the bio-protective agent loaded biological material for storage; storing the biological material; recovering the stored biological material from storage; and reversing the cell membrane poration. H5 ?-toxin, a genetically engineered mutant of Staphylococcus aureus ?-hemolysin, may be used as a porating agent. Non-permeating sugars such as trehalose and sucrose may be used as the bio-protective agent.

Abstract: The present invention provides a method for creating a frozen tissue array. An oil in a liquid form is added into a recipient block containing frozen tissue cores where the oil has a freezing point lower than the freezing point of the tissue cores; and the recipient block containing the oil is cooled to a temperature about equal to or below the freezing point of the oil; the subsequently frozen oil locks the frozen cores in the recipient block. The oil may also be added to the recipient block prior to inserting the frozen tissue cores. The recipient block may be formed using a cryoarray device comprising a mold plate, an ejector plate, mold alignment pins, ejector pins, and cryoarray pins. Such method may be used for preparing frozen sections with multiple tissue specimens for assays such as in-situ hybridization and immunohistochemistry.

Abstract: This invention relates to formulations and methods for preserving sensitive biologicals, viruses, bacteria and eukaryotic cells by drying. More particularly, the invention relates to preservation mixtures containing viruses or cells and protectants, including a combination of a methylated monosaccharide and a disaccharide, or oligosaccharide, wherein the mixtures are adapted to stabilize these during dehydration and subsequent storage at ambient and higher temperature.

Abstract: Novel methods and devices for removing cryoprotectant from cryoprotectant-containing liquids, and from cells residing therein, are disclosed. In one aspect, the method comprises passing the cryoprotectant-containing liquid through at least one semipermeable hollow fiber membrane contained in a hollow module in a first direction, while passing a liquid which is substantially free of cryoprotectant through the hollow module in a second direction to remove cryoprotectant across a diffusion gradient. In another aspect, a device is described for removing cryoprotectant from a liquid, comprising a hollow module with at least one semipermeable hollow fiber membrane therein for accomplishing such counter-current diffusion removal of cryprotectant. A software program is also provided for predicting optimal flow rates through the device of the invention, thereby allowing optimal cryoprotectant removal regardless of the cryoprotectant used or the material from which the semipermeable hollow fiber membrane is fabricated.

Abstract: Disclosed herein is a carrier solution for cryoprotectants that is useful for use with cells, tissues, and whole organs and for a variety of cryoprotectant solutions and that permits antinucleators to be fully effective in vitrification solutions, thereby allowing vitrification solutions to attain extreme effectiveness, and compatible vitrification solution compositions for use with this carrier solution. The carrier solution comprises lactose and mannitol as well as other beneficial ingredients.