Abstract: A cryoprotective device protects an aqueous biological material from mechanical damage due to ice formation during cryogenic freezing and/or cryostorage by preventing direct contact of the biological material with cell-damaging large ice crystals, the cryoprotective storage device having a housing with an internal cavity. The housing is configured to receive a freezable medium with the biological material within the internal cavity. The housing includes a semi-permeable membrane. The membrane is impermeable to ice crystals that are larger than an average pore size of the membrane to prevent such ice crystals from passing into the internal cavity from outside the housing, such that ice crystals formed in the medium within the housing have a smaller crystal size from ice crystals formed in the medium outside of the housing. As such, the biological material is protected from mechanical damage generated by direct contact with large ice crystals.

Abstract: An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane that is impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. One such cryopreservation vessel is a flexible vial having a closed port at one adapted to be pierced by a needle to withdraw the liquid within, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. In another embodiment, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Abstract: An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane that is impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. One such cryopreservation vessel is a flexible vial having a closed port at one adapted to be pierced by a needle to withdraw the liquid within, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. In another embodiment, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Abstract: An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. The cryopreservation vessel can be a flexible vial having a closed port adapted to be pierced by a needle, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. Alternatively, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Abstract: A method for cryopreservation of animal cells with high level of intracellular lipid content, comprises the steps of conducting a delipation procedure using one or more lipolytic agent(s) and/or lipogenesis inhibitors during culture of the animal cells to stimulate the hydrolysis of intracellular lipids to reduce the lipid content, and vitrifying the treated animal cells using a modified vitrification solution and a modified warming solution.

Abstract: An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. The cryopreservation vessel can be a flexible vial having a closed port adapted to be pierced by a needle, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. Alternatively, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Abstract: An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane that is impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. One such cryopreservation vessel is a flexible vial having a closed port at one adapted to be pierced by a needle to withdraw the liquid within, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. In another embodiment, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Abstract: Methods are disclosed in which the expression of a specific gene, or combinations of genes, is controlled spatially and temporally to develop intra- and interspecies chimeras. A transgenic EC/ES/P/iPS cell line is created which conditionally expresses a suicide or compromiser gene configured to compromise all cell lineages except that corresponding to a target tissue/organ. The EC/ES/P/iPS cell line is injected into donor embryos having a specific target gene deficiency or embryos genetically engineered to be complementary compromised in lineages corresponding to the target tissue/organ cell lineages of the EC/ES/P/iPS line. One or more stimuli is provided to the embryo to activate compromiser genes for ablation of non-target tissues/organs of the EC/ES/P/iPS line and target tissues/organs of the host embryo, resulting in a chimeric animal having target tissues/organs derived from the genotype of the transgenic cell line and all remaining tissues/organs derived from the donor embryo.

Abstract: An auto-nucleating device includes a hollow tube containing a crystalline cholesterol matrix therein. The ends of the tube are closed by a membrane that is impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device is disposed within the vessel and provides a site for ice nucleation during freezing of the liquid within the vessel. One such cryopreservation vessel is a flexible vial having a closed port at one adapted to be pierced by a needle to withdraw the liquid within. The opposite end of the vial is initially open to receive the liquid. The end is then sealed to form a closed system for cryopreservation. Another cryopreservation vessel includes an adaptor mounted to a port of a liquid container. The adaptor includes one tubular branch that is closed by a needle septum while another tubular branch includes a barbed fitting for engaging a flexible tube. The flexible term terminates in a needle septum.

Abstract: A device and method suitable for the cryopreservation of all types of biological cells is described. In this method, an ultra-fast cooling/warming device system is used to achieve vitrification of individual cells or cell suspensions without cryoprotectant agents (CPA) or with a low concentration of CPAs (<1M), to attenuate the formation of intracellular ice crystal formation during cooling, and to minimize devitrification during subsequent warming. The device system applies oscillating heat pipe (OHP) and nanofluid techniques, and is built through microfabrication. Several devices may be networked to increase the total volume of cell samples that the cryopreservation system can process simultaneously.

Abstract: A method for cryopreservation of animal cells with high level of intracellular lipid content, comprises the steps of conducting a delipation procedure using one or more lipolytic agent(s) and/or lipogenesis inhibitors during culture of the animal cells to stimulate the hydrolysis of intracellular lipids to reduce the lipid content, and vitrifying the treated animal cells using a modified vitrification solution and a modified warming solution.

Abstract: Methods are disclosed in which the expression of a specific gene, or combinations of genes, is controlled spatially and temporally to develop intra- and interspecies chimeras. A transgenic EC/ES/P/iPS cell line is created which conditionally expresses a suicide or compromiser gene configured to compromise all cell lineages except that corresponding to a target tissue/organ. The EC/ES/P/iPS cell line is injected into donor embryos having a specific target gene deficiency or embryos genetically engineered to be complementary compromised in lineages corresponding to the target tissue/organ cell lineages of the EC/ES/P/iPS line. One or more stimuli is provided to the embryo to activate compromiser genes for ablation of non-target tissues/organs of the EC/ES/P/iPS line and target tissues/organs of the host embryo, resulting in a chimeric animal having target tissues/organs derived from the genotype of the transgenic cell line and all remaining tissues/organs derived from the donor embryo.

Abstract: An auto-nucleating device includes a tube containing a crystalline cholesterol matrix. The ends of the tube are closed by a membrane that is impermeable to the cholesterol but permeable to liquids contained in a cryopreservation vessel. The auto-nucleating device provides a site for ice nucleation during freezing of the liquid within the vessel. One such cryopreservation vessel is a flexible vial having a closed port at one adapted to be pierced by a needle to withdraw the liquid within, and an opposite end that is initially open to receive the liquid. Another vessel includes an adaptor mounted to liquid container with a tubular branch closed by a needle septum and another tubular branch provided with a barbed fitting for engaging a flexible tube that terminates in a needle septum. In another embodiment, the vessel includes an inlet and vent branch at the top of the container and an outlet septum at a bottom opening.

Abstract: A method to optimize a vitrification procedure for suspended cells uses factors such as the physical properties of solutions, the cell permeability to water and permeable cryoprotectants, and the osmotic tolerance of the cells to identify a method to minimize several stresses associated with vitrification procedures.

Abstract: A method is disclosed for removal of cryoprotectants from preserved suspension of biological cells and tissues. Sorbent materials are used, alone or in combination, to bind the cryoprotectant component of preserved cell suspensions with minimal osmotic stress on the preserved cells. The present method is used to effectively remove the cryoprotectants from cryopreserved cells and tissues prior to their use in transfusion, transplantation, insemination or other applications, with minimal osmotic damage due to cell swelling. Specific devices and methods are described.

Abstract: A method to remove cryoprotectants from cryopreserved biological cells and tissues is described. Enzymes are used to convert compounds used as cryoprotectants for cells during cryopreservation to membrane impermeable products. This process effectively removes the cryoprotectant chemicals from the cells prior to their use in transfusion, transplantation, insemination or other applications, without causing osmotic damage due to cell swelling. Methods are described for subsequent removal of the enzymes and enzyme conversion products from the cell and tissue preparations.

Abstract: A device for analyzing the in-vitro growth properties of cells is disclosed. The device comprises an upper and lower chamber separated by a growth substrate interface comprising submucosa, preferably tunical submucosa, from a warm-blooded vertebrae. Methods for culturing eukaryotic cells and studying their growth characteristics, including the invasive growth characteristics of tumor cells, on the submucosal matrix are described.

Abstract: A mathematical model, the membranes and devices based upon that model to optimize protocols for the addition or removal of cryoprotectant to or from biological cells, and a method to observe the biological cells and obtain the data to implement the models. This disclosure describes the use of four equations to predict optimal protocols to add or remove cryoprotectant to or from biological cells. The equations particularly require experimentally found data, specific to cell-type and species, regarding the osmotic tolerance of the cells, where osmotic tolerance refers to the cells ability to shrink or swell to various changes in osmolality without injury. The equations further require the cryoprotectant permeability coefficient and the water permeability coefficient of the particular cells' plasma membrane.

Abstract: A mathematical model, and membranes and devices based upon that model, to optimize protocols for the addition or removal of cryoprotectant to or from biological cells. This disclosure describes the use of four equations to predict optimal protocols to add or remove cryoprotectant to or from biological cells. The equations particularly require experimentally found data, specific to cell-type and species, regarding the osmotic tolerance of the cells, where osmotic tolerance refers to the cells ability to shrink or swell to various changes in osmolality without injury. The equations further require the cryoprotectant permeability coefficient and the water permeability coefficient of the particular cells' plasma membrane. These coefficients are found with experimental data of the knetic volume change of the cell-type to a known concentration and temperature of cryoprotectant.

Abstract: A mathematical model, and membranes and devices based upon that model, to optimize protocols for the addition or removal of cryoprotectant to or from biological cells. This disclosure describes the use of four equations to predict optimal protocols to add or remove cryoprotectant to or from biological cells. The equations particularly require experimentally found data, specific to cell-type and species, regarding the osmotic tolerance of the cells, where osmotic tolerance refers to the cells ability to shrink or swell to various changes in osmolality without injury. The equations further require the cryoprotectant permeability coefficient and the water permeability coefficient of the particular cells' plasma membrane. These coefficients are found with experimental data of the knetic volume change of the cell-type to a known concentration and temperature of cryoprotectant.