607 results about "Cryoprotectant" patented technology

The present invention is in the area of administration forms and delivery systems for drugs, vaccines and other biologically active agents. More specifically the invention is related to the preparation of suspensions of colloidal solid lipid particles (SLPs) of predominantly anisometrical shape with the lipid matrix being in a stable polymorphic modification and of suspensions of micron and submicron particles of bioactive agents (PBAs); as well as to the use of such suspensions or the lyophilizates thereof as delivery systems primarily for the parenteral administration of preferably poorly water-soluble bioactive substances, particularly drugs, and to their use in cosmetic, food and agricultural products. SLPs and PBAs are prepared by the following emulsification process: (1) A solid lipid or bioactive agent or a mixture of solid lipids or bioactive agents is melted. (2) Stabilizers are added either to the lipid or bioactive agent and to the aqueous phase or to the aqueous phase only depending on their physicochemical characteristics. Stabilizers may also be added or exchanged after homogenization. (3) Drugs or other bioactive substances to be incorporated into the SLPs may be melted together with the lipids if the physicochemical characteristics of the substance permit or may be dissolved, solubilized or dispersed in the lipid melt before homogenization. (4) The aqueous phase is heated to the temperature of the melt before mixing and may contain for example stabilizers, isotonicity agents, buffering substances, cryoprotectants and/or preservatives. (5) The molten lipid compounds and the bioactive agents are emulsified in an aqueous phase preferably by high-pressure homogenization.

The present invention comprises the discovery and development of an effective cryoprotectant composition, without containing skim milk or any other animal-derived compounds, to achieve long-term stability of freeze-dried lactic acid bacteria (LAB), at different temperatures, whereby the retention of viability of the freeze-dried LAB after 6 months of storage, preferably after 9 months of storage, more preferably after 12 months of storage is more than 50%. The invention is in the field of producing freeze dried bacteria, in particular Lactic acid bacteria. More in particular, the invention relates to the use of a novel combination of cryoprotectants for increasing the viability of bacteria after freeze drying, improving the texture of the lyofilized cake for easy grinding and improving the long term stability of the freeze dried bacteria at different temperature conditions. The invention further relates to such freeze dried bacteria for use in food industry or in human or animal health applications. More in particular, the invention relates to the increased viability and long-term storage of recombinant bacteria capable of expressing heterologous proteins or peptides and administered to humans or animals for therapeutic or vaccination purposes.

A cryopreservation medium and a cryopreservation method which make it possible to cryopreserve ES cells from primates simply with high viability are provided. A cryopreservation medium containing a cryoprotectant at a Concentration of from 12% (W/V) to 50% (W/V) and a cryopreservation method for primate embryonic stem cells, which comprises a step of suspending primate embryonic stem cells in the cryopreservation medium, and a refrigeration step of freezing the suspension of the primate embryonic stem cells in the cryopreservation medium by cooling it to −80° C. or below at a rate of from 0.5° C. to 10° C. per minute, and a preservation step of storing the frozen suspension of primate embryonic stem cells in the cryopreservation medium enable simple cryopreservation of primate embryonic stem cells with high viability.

The inventive technology relates to methods and apparatus for reducing protein content in sperm cell extenders and may include one or more of the following features: techniques for reducing protein content in a sperm cell extender; techniques for reducing protein content in a cryoprotectant-containing B fraction of a sperm cell extender; techniques for preparing sperm cell extenders that do not require clarification; techniques for preparing low density gradient sperm cell extenders suitable for centrifugation; techniques for reducing protein content between individual steps in preparing a sperm cell extender, and techniques for establishing novel values of reduced protein content in sperm cell extenders.

The invention relates to the field of stem cells, discloses a method for preparing exosome freeze-dried powder of stem cells, and particularly relates to a method for preparing exosome freeze-dried powder of human amniotic mesenchymal stem cells. The method for preparing the exosome freeze-dried powder of the human amniotic mesenchymal stem cells disclosed by the invention comprises the following steps: mixing trehalose with exosomes of the human amniotic mesenchymal stem cells as a cryoprotectant; filtering and removing bacteria by virtue of a filter membrane; and firstly carrying out ultralow temperature pre-freezing, and then carrying out low-temperature freezing at a certain vacuum degree. The preparation method disclosed by the invention is simple to operate, safe and effective. An experiment proves that compared with a conventional preparation method of the freeze-dried powder, the morphologies and the activity of the exosomes of the human amniotic mesenchymal stem cells can be well kept by the method for preparing the exosome freeze-dried powder of the human amniotic mesenchymal stem cells disclosed by the invention; and the method is suitable for long-term preservation and application of the exosomes of the human amniotic mesenchymal stem cells.

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. Tissues or organs treated in this manner exhibit near normal functions, for example, blood vessels exhibit near normal smooth muscle contractility and normal graft functions.

Provided herein are use of polymeric excipients, specifically polyvinyl alcohols, optionally in conjunction with sugars, as cryoprotectants to prevent aggregation of PEG-containing particles. Also provided are PEG-containing particles comprising such polymeric excipients.

Cultures of keratinocyte cells are provided which are free from nonautologous fibroblasts and organ extracts, and which have a high speed of cell amplification for a minimum seeding density. The cultures can be cryopreserved in a buffered isotonic medium containing serum and a cryoprotectant. The cultures are produced by a process that does not involve the use of a feeder layer and organ extracts. A culture medium which can be used contains Medium 199, serum, epidermal growth factor, cholera toxin and/or hydrocortisone, and optionally insulin. A substance for wound healing and for cosmetic applications is derived from cultured human keratinocytes. A non-viable total keratinocyte lysate for use in promoting wound healing is produced by growing keratinocyte cells on a support, detaching the cells from the support, and lysing the detached cells to obtain the lysate which may be frozen and lyophilized. The cells may be grown without using a support to produce the lysate, or to produce a non-viable keratinocyte cell culture lyophilisate or spray dried non-viable keratinocyte cell composition for use in healing wounds.

Acoustophoretic devices are disclosed. The devices include a flow chamber, an ultrasonic transducer, a reflector, an inlet, a filtrate outlet, a concentrate outlet, and optionally a lipid collection trap. The ultrasonic transducer and reflector create a multi-dimensional acoustic standing wave in the flow chamber that traps and separates red blood cells and/or lipids from blood. Concentrated red blood cells can be recovered via the concentrate outlet, the lipids can be recovered via the lipid collection trap, and the remaining blood can be recovered via the filtrate outlet. Methods for separating blood components (e.g., red blood cells, lipids, platelets, white blood cells) from blood are also disclosed. The red blood cells can undergo washing with a solvent to remove undesired admixtures. Cryoprotectants can be added or removed from the blood.

A non-linear cooling cryopreservation method for improving cryopreservation protocols for cells that involves producing a simulation of cellular responses to a range of cooling parameters; determining optimal cooling parameters required to minimize cryoinjury to the cells using simulation of cellular responses and experimental results; and incorporating optimal parameters into the protocol. The simulation is based on mathematical models of cellular parameters. A non-linear cooling cryopreservation protocol for cryopreserving stem cells is also disclosed that does not require cryoprotectants.

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.

A method for cryopreservation of cells characterized by the use of a solution containing, as essential components, a sodium salt, a potassium salt, a saccharide, a cryoprotectant, and a hydrogencarbonate salt and/or a carbonate salt, the solution further optionally containing a component selected from the group consisting of proteins, magnesium salts, and calcium salts. According to the present invention, a method for cryopreservation of cells capable of maintaining a high viability rate after thawing, and a cryopreservation solution used in the cryopreservation are provided. In addition, by using the method for cryopreservation, the cells can be preserved in a state of maintaining a high viability rate even after thawing, and also maintaining their physiological functions; further, a cell washing step after freezing and thawing is unnecessary, thereby making it very useful in the fundamental researches of cells and the application studies on the medical treatment.

The invention relates to a method for preparing a freeze-dried microbial agent of kluyveromyces marxianus and application of the freeze-dried microbial agent. The freeze-dried microbial agent is prepared by the aid of high-density fermentation culture and freeze-drying technologies. The kluyveromyces marxianus is preserved in the Common Microorganism Center of the China Committee for Culture Collection of Microorganisms, called as CGMCC for short, on April 27th, 2010, the preservation number of the kluyveromyces marxianus is CGMCC NO.3781, and the strain number is A3. The method and the application have the advantages that the viable cell number of high-density culture solution of the kluyveromyces marxianus can reach 3.2*10<8> cfu/mL at least and is higher than culture results of YEPD (yeast peptone dextrose) culture media by 1.1 order of magnitude at least; zymocyte solution is centrifuged, then bacterial sludge is collected and is added into cryoprotectant solution, the freeze-dried microbial agent can be prepared by the aid of the vacuum freeze-drying technologies, protectants for the freeze-dried microbial agent comprise maltose and skimmed milk powder, and the freeze-dried viable bacterium rate can reach 75% at least; the freeze-dried microbial agent is high in bacterium content, long in storage time, convenient to use and free of pollution, is environmentally friendly and can be used for fermenting milk beer and cheese and making whey wine, fruit vinegar, fruit juice and the like, processes for preparing the freeze-dried microbial agent are simple, and the like.

The invention relates to cryopreservation protection liquid for Wharton jelly tissues of a human umbilical cord. The cryopreservation protection liquid comprises 5-10 percent of a permeable cryoprotectant, 1-5 percent of a non-permeable cryoprotectant, 10-20 percent of a knockout serum replacement and 70-80 percent of a serum-free basal culture medium. The invention also discloses a preparation method of the cryopreservation protection liquid. The preparation method specifically comprises the steps of (1) precooling the serum-free basal culture medium, the non-permeable cryoprotectant and the knockout serum replacement, mixing and uniformly shaking; (2) adding the permeable cryoprotectant; and (3) performing refrigeration under a temperature condition of 2-6 DEG C for more than 30min. The invention also discloses a cryopreservation method for the Wharton jelly tissues of the human umbilical cord by the cryopreservation protection liquid. The cryopreservation method comprises the following steps of (1) pretreating the umbilical cord; (2) peeling off the Wharton jelly tissues; and (3) cryopreserving the Wharton jelly tissues of the human umbilical cord. The cryopreservation protection liquid and the cryopreservation method which are used for treating the Wharton jelly tissues of the human umbilical cord have the advantages that injury to umbilical cord mesenchymal stem cells in a cryopreservation process can be alleviated, so that the activity and the clinical use safety of the umbilical cord mesenchymal stem cells are guaranteed.

The present invention features novel methods for the cryopreservation of mammalian cell that combine the advantages of the slow-freezing and vitrification approaches while avoiding their shortcomings. Generally, the methods include the use of a capillary tube made of a thermally conductive wall material and a thin wall such that the ratio of the thermal conductivity of the wall material to the wall thickness is at least 1,000-500,000. The solution is then exposed to temperatures equal to or less than −80° C. and the vitrification solution containing the mammalian cells is cooled at a rate equal to or greater than 30,000-100,000,000° C./minute. The exposure of the capillary tube with a thermally conductive and thin wall allows for vitrification of the solution in the absence of ice formation. Cryoprotectants can also be added to the vitrification solution to further prevent ice formation.