Methodist Hospital of Indiana, Inc.

Cook General Biotechnology LLC

Purdue Research Foundation

University of Missouri

General Biotechnology, LLC

Clarian Health Partners, Inc.

Bio Technology, LLC

Critser, John K.

Woods, Erik John

Methodist Health Group, Inc.

John K Critser

Erik John Woods

Robert J Demeter

Dayong Gao

D Y Gao

Stephen Francis Badylak

Sherry L Voytik-Harbin

George B Boder

Chi Liu

Peter Koulen

Xu Han

Hongsheng Men

Sherry Voytik

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.

Efficient cryopreservation device preventing the direct contact between samples and extracellular ice

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.

Systems and methods for cryopreservation of cells

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.

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.

Methods for the cryopreservation of animal cells that contain high levels of intracellular lipids

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. The container is initially at below-atmospheric pressure. In use, the liquid sample is injected by needle through the septum in the second branch. Once the container is filled with the sample, the flexible tube is heat sealed and severed just above the barbed fitting. The sealed container may then be subject to a cryopreservation protocol. After thawing, the sample liquid may be withdrawn by a needle puncturing the septum in the first branch.

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.

Sorbent method for removal of cryoprotectants from cryopreserved animal cells

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.

Enzymatic method for removal of cryoprotectants from cryopreserved animal cells

Device and method for analyzing tumor cell invasion of an extracellular

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. These coefficients are found with experimental data of the knetic volume change of the cell-type to a known concentration and temperature of cryoprotectant, and one method is particularly presented as to how this data may be obtained. Also disclosed are particularly preferred methods and devices to add or remove cryoprotectant to or from these cells based upon these equations.

Cell-type-specific methods and devices for the low temperature

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. Also disclosed are particularly preferred methods and devices to add or remove cryoprotectant to or from these cells based upon these equations.

Cell-type specific methods and devices for the low temperature

General method to quickly add cryoprotectants to animal cells while

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. Also disclosed are particularly preferred methods and devices to add or remove cryoprotectant to or from these cells based upon these equations.

General method to quickly remove cryoprotectants from animal cells while

A cell culture growth substrate comprising submucosal tissue of a warm-blooded vertebrate and a method for culturing eukaryotic cells are described. Submucosal tissue used in accordance with the present invention supports the proliferation and differentiation of eukaryotic cells when said cells are contacted with submucosal tissue under conditions conducive to cell proliferation.

Submucosa as a growth substrate for islet cells

A mathematical model to optimize protocols for the addition or removal of cryoprotectant to or from sperm cells. This disclosure describes the use of four equations to predict optimal protocols to add or remove cryoprotectant to or from sperm cells. The equations particularly require experimentally found date, specific to a species of sperm, regarding the osmotic tolerance of sperm cells, where osmotic tolerance refers to the sperm 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 sperm cells' plasma membrane. Also disclosed are two particularly preferred methods to add or remove cryoprotectant to or from sperm. These preferred methods are Fixed-Volume-Step addition/removal of cryoprotectant or Fixed-Molarity-Step addition/removal of cryoprotectant.

Growing community of inventors

Columbia, MO, United States of America

John K Critser