147 results about "Cryopreserved Cell" patented technology

The present invention is directed to methods of cryopreserving cells and cryopreserved cells prepared according to the methods. In specific embodiments, the method comprises combining cells with a cross-linked hydrogel matrix in particulate form, the matrix comprising a polyglycan cross-linked to a polypeptide and subjecting the combination to cryopreservation conditions. In further embodiments, the invention provides cell-seeded compositions comprising cells and a cross-linked bioactive hydrogel matrix in particulate form, the matrix comprising a polyglycan cross-linked to a polypeptide, wherein the composition has been subjected to cryopreservation conditions. The cryopreserved cells can be thawed and used in methods of treatment without the need for intervening steps to make the cells viable for in vivo use.

The present invention relates to methods for cryopreserving plant cells and to methods for recovering viable plant cells from long or short term cryopreservation. Plant cells to be cryopreserved can be grown in culture and pretreated with a solution containing an cryoprotective agent and, optionally, a stabilizer. Stabilizers are preferably membrane stabilizers such as ethylene inhibitors, oxygen radical scavengers and divalent cations. Cells can also be stabilized by subjecting the culture to a heat shock. Pretreated cells are acclimated to a reduced temperature and loaded with a cryoprotective agent such as DMSO, propylene glycol or polyethylene glycol. Loaded cells are incubated with a vitrification solution which, for example, comprises a solution with a high concentration of the cryoprotective agent. Vitrified cells retain less than about 20% water content and can be frozen at cryopreservation temperatures for long periods of time without significantly altering the genotypic or phenotypic character of the cells. Plant cells may also be cryopreserved by lyophilizing cells prior to exposure to a vitrification solution. The combination of lyophilization and vitrification removes about 80% to about 95% of the plant cell's water. Cells can be successfully cryopreserved for long periods of time and viably recovered. The invention also relates to methods for the recovery of viable plant cells from cryopreservation. Cells are thawed to about room temperature and incubated in medium containing a cryoprotective agent and a stabilizer. The cryoprotective agent is removed and the cells successfully incubated and recovered in liquid or semi-solid growth medium. The invention also relates to the cryopreserved cells and to viable plant cells which have been recovered from long or short term cryopreservation.

The invention relates to the field of biological medicines and in particular to cocktail type cryoprotectant and application thereof. The cocktail type cryoprotectant comprises the following components: 1-60% (w / v) of a polysaccharide, 1-50% (w / v) of an amino acid, 1-20% (w / v) of a polyhydric alcohol and 0-10% (w / v) of a hydroxyl-containing macromoleclar polymer. The polysaccharide, the amino acid, the polyhydric alcohol and the hydroxyl-containing macromoleclar polymer of the cryoprotectant are relatively low in toxicity with high concentrations, formation of intracellular ice crystals can beeffectively inhibited with the polysaccharide, the amino acid, the polyhydric alcohol and the hydroxyl-containing macromoleclar polymer of the proportions, cryopreserved cells can be alleviated fromdamage caused by low temperature environments, the survival rates of the cryopreserved cells after anabiosis culture can be increased, the toxicity of the cryopreserved cells can be greatly reduced, and thus the cocktail type cryoprotectant provided by the invention has very high security feasibility and application and popularization values.

A composition and method for cryopreserving cells in a non-toxic medium is described. The composition and method provide cells for cell therapy and other in vivo applications.

The invention discloses a cryoprotective agent of peripheral blood mononuclear cells. The cryoprotective agent is prepared from the following ingredients in parts by weight: 43-53 parts of a culture medium, 30-40 parts of auto-plasma, 12-17 parts of dimethyl sulfoxide, 5-7% of human serum albumin, and the balance being 0.17-0.23mol / L trehalose. The invention discloses a preservation method of the cryoprotective agent. The preservation method comprises the steps of collecting fresh anticoagulation blood, preparing peripheral mononuclear cells into cell suspension by using human serum albumin, adding the cryoprotective agent with the same volume as that of the cell suspension, mixing uniformly, preserving for 10-20h at below 90-below-70-below DEG C, and preserving in liquid nitrogen. According to the cryoprotective agent and the preservation method thereof, a cryopreserved cell has the high motility rate and the strong cell activity and can shorten the Cytokine Induced Killer (CIT) cell induced amplification period.

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.

The invention relates to a gel preparation for maintaining the activity of cryopreserved cells and a stem cell gel preparation containing stem cells. The gel preparation comprises the following components by the mass percentage: 1-3% of sodium alginate, 1-5% of dimethyl sulfoxide, 1-5% of propylene glycol, 1-5% of epigallocatechin gallate, 2-10% of dextran, 1-5% of human blood albumin, and water or a phosphate solution added to make the total amount be 100%. The gel preparation has the advantages of good biocompatibility and simple operation, can maintain high activity of cells, can be directly used for low temperature preservation, and has no need of traditional liquid nitrogen cryopreservation; the cell gel preparation after resuscitation can still maintain high activity of the cells and dryness of the cells, and can be applied for treatment of skin injury or mucosal injury.

The present invention relates to methods and structures for preparing stem cells for use in cryopreservation methods. Stem cell colonies are provided between first and second matrix portions and are exposed to a carbohydrate-containing cryoprotecting medium and a freezing medium. The methods of the invention yield cryopreserved cells that maintain cell viability and exhibit limited cell differentiation after freezing and thawing, facilitating storage, shipping and handling of embryonic stem cell stocks and lines for research and therapeutics.

A method of preparing and thawing of cryopreserved cells without added DNase and a method of DNase-free isolation of subpopulations of thawed, cryopreserved cells which can be used to prepare and thaw Human Cord Blood cells for immunoaffinity selection and enrichment of CD34+ hematopoietic progenitor cells for expansion and transfusion.

The invention relates to compositions for the cryogenic storage of biological materials and related methods. In an embodiment, the invention includes a cryopreservative composition including a chaotropic agent and a kosmotropic agent. In an embodiment, the invention includes a cryopreservative composition including urea and trimethylamine-N-oxide. In an embodiment, the invention includes a method of cryopreserving cells including contacting cells with a cryopreservative composition, the cryopreservative composition comprising a chaotropic agent and a kosmotropic agent. In an embodiment, the invention includes a method of transplanting cells into a subject, the method including administering a composition to the subject, the composition comprising an effective amount of a chaotropic agent, an effective amount of a kosmotropic agent, and cells. Other embodiments are also included herein.

This invention relates to methods and apparatus for cryopreserving biological materials for extended periods of time. In an exemplary embodiment, the method comprises suspending biological materials in a cryosolution, freezing the biological materials in the apparatus, and removing the frozen biological materials from the apparatus to thaw them for use. In another embodiment, a cell cryopreservation solution is provided which includes 20% Dimethyl Sulfoxide (DMSO) to maintain the viability of cells upon freezing, storage, and thawing. The media can be used for cryopreservation of a wide variety of different cell types from various sources. In addition, an apparatus that facilitates the storage of cells and the subsequent removal of the cells for thawing to permit substantially direct inoculation of a bioreactor is disclosed. Cells frozen using a method according to the present disclosure have been shown to have approximately a 90% survival rate, which is significantly higher than other cryopreservation methods.

The invention provides cell cryopreservation liquid and a preparation method thereof. The cell cryopreservation liquid is prepared from, 8% w / v of dimethyl sulfoxide, 30 ng / ml of total flavonoids of herba epimedii, 1% w / v of polypeptide, 1% w / v of bFGF and 1% w / v of vitamin E. The cell cryopreservation liquid is used for cryopreserving culture cells, and especially the survival rate of the recovered cryopreserved cells is increased. The cell cryopreservation liquid has the advantages of being moderate in price and easy and convenient to operate and is suitable for being widely utilized.

Disclosed herein are methods for cryopreserving cells and tissues under clinical conditions, allowing production of viable cell products suitable for transplantation.

The invention relates to a cryopreservation solution, a cryopreservation method and a recovery method for peripheral blood mononuclear cells. The cryopreservation solution includes, by volume, 40-60%of a serum-free medium, 15-25% of an impermeable protective agent, 5-15% of a permeable protective agent, 5-15% of sodium hyaluronate and 5-15% of an anti-oxidation substance. The cryopreservation solution composed of the serum-free medium, the impermeable protective agent, the permeable protective agent and the anti-oxidation substance can effectively cryopreserve cells, and makes the cryopreserved cells still have a high cell viability after being recovered, so the cryopreservation solution has high application values, and is worth promoting.

A method for cryopreserving cells entails the liposomal delivery of intracellular sugar(s), such as trehalose, sucrose, raffinose, stachyose, and combinations thereof, into cells and tissues, such as red blood cells, for enhancing post-thaw viability. This method enables rapid and easy delivery of protective molecules into cells which thus greatly simplifies the preparation of cells for cryopreservation. Furthermore, as much lower concentrations of intracellular protectant are used, the method allows red blood cells containing the liposomally-delivered intracellular sugar to be transfused into a patient immediately following the thaw without having to first remove any of the cryoprotectant sugar.

Provided are apparatuses for cryopreserving cells which include a vessel comprising a biocompatible substrate, wherein the vessel further comprises an interior and an exterior, and a mechanical ice nucleating device disposed in or on the vessel interior for initiating ice crystal formation. Also provided are kits comprising one or more apparatuses for cryopreserving cells and a biopreservation medium. Further provided are compositions comprising a vessel for holding cells, a mechanical ice nucleating device, a biopreservation medium, and cells disposed in the vessel. The apparatuses, kits, and compositions of the invention can optionally include an insulating material which is disposed on all or a portion of the vessel.

The invention relates to the field of cells, in particular to a cell cryoprotectant and a cryopreservation method. The cell cryoprotectant is prepared from DMSO, human albumin and a serum-free medium. The cryprotectant does not contain animal serum, the risks of introducing contamination and allergens are avoided, and the higher clinical safety is achieved compared with a conventional cell cryoprotectant. Meanwhile, the GMSCs cryoprotectant can well keep the activity of cryopreserved cells.

The invention discloses a method for long-term storage and resuscitation culture of adult peripheral blood mononuclear cells, and relates to a separation, cryopreservation and resuscitation method of adult peripheral blood mononuclear cells. The method comprises the following steps: I, collecting peripheral venous blood of a healthy adult by using a blood taking bag, and separating and purifying the mononuclear cells in the blood; II, adding related cryopreservation reagents necessary for cryopreservating cells so as to cryopreservate the separated mononuclear cells; III, randomly sampling, and detecting bacteria, fungi, endotoxin and mycoplasma in a cell cryopreservation mixed liquid; and IV, resuscitating the cells, suspending in specially prepared cell resuscitation lotion, performing centrifugal washing twice, and further culturing. As the peripheral blood mononuclear cells used in the method are taken from a patient self, the related reagents used in the test are free of exogenous material, related ethical issues can be avoided, and immunological rejection can be avoided after the cells are injected back to the body. The method aims to store healthy adult immune cells for later treatment on diseases related to tumor.

The invention relates to a device and a method for cryopreserving animal cells in batch; the device consists of a cell cryopreservation bag, a cell cryopreservation box and a cell rack; the cell cryopreservation bag is positioned on the cell rack; and the cell rack is positioned inside the cell cryopreservation box. The method for cryopreserving the cells in batch comprises the following steps that: a cell suspension is filled in the cell cryopreservation bag and positioned on the cell rack; the cell rack is put in the cell cryopreservation box; the cell cryopreservation box is thrown into a liquid nitrogen tank or freezing equipment at a temperature of less than 70 below zero for freezing 24 hours; the cell cryopreservation bag is taken out, thrown into liquid nitrogen or the freezing equipment at the temperature of less than 70 below zero and cryopreservated for a long time; when the cells need to be defreezed, the cell cryopreservation bag is pulled out and put in warm water at a temperature of between 38 and 39 DEG C and defreezed; after coagulum is molten into liquid, the liquid is moved to a laminar flow hood; according to a sterile operating procedure, the cell suspension in the cell cryopreservation bag is subpackaged into a centrifuge tube for centrifugation; a supernatant fluid is discarded; and the suspension is operated according to routine cell cryopreservation. The volume of the cryopreserving cell of the cryopreservation bag reaches between 50 and 200 ml; and the whole set of the device is directly thrown into the liquid nitrogen or put in the freezing equipment at the temperature of less than 70 DEG C below zero.

The invention belongs to the field of cell cryopreservation and particularly relates to a periodontal ligament stem cell cryoprotectant and a cryopreservation method thereof. The cryoprotectant disclosed by the invention is prepared from glycerinum, mesenchymal steam cell serum-free medium and human serum albumin. The cryoprotectant disclosed by the invention can effectively solve the technical defects that an existing cryoprotectant has low use effect and a survival rate of post-resuscitation periodontal ligament stem cells is low. The periodontal ligament stem cell cryoprotectant disclosed by the invention can keep activity of cryopreserved cells; furthermore, the survival rate of the post-resuscitation periodontal ligament stem cells is obviously improved, and expression of surface markers of the stem cells and differentiation potential of the stem cells are not affected.

The invention relates to a cryoprotectant based on liquid metal nanoparticles as well as a method and an application of the cryoprotectant. The cryoprotectant comprises the liquid metal nanoparticlesand a basic protectant, wherein the melting point of the raw material of the liquid metal nanoparticles is below 60 DEG C, and the basic protective agent is a mixture of ethylene glycol, propylene glycol and trehalose. The cryoprotectant can realize vitrification in the cooling process, and can be used as a space heat source under the irradiation of near-infrared laser in the rewarming process, thereby greatly enhancing the heating rate, inhibiting anti-vitrification, reducing the formation of ice crystals, and enhancing the survival rate and structural integrity of cryopreserved cells and vascular tissues.

The invention provides a method for quickly cryopreserving and thawing cells, which comprises a cryopreservation step and a thawing step. The cryopreservation step comprises: (1) cells are dissociated, and cell suspension is prepared; (2) the cells are diluted and separately packaged; (3) temperature is gradiently decreased, and is increased, temperature is then gradiently decreased to negative 120 DEG C to negative 180 DEG C, and the cells are stored in liquid nitrogen. The thawing step comprises: the cryopreserved cells are taken out of the liquid nitrogen, quickly immersed in thawing waterwhich is 38 DEG C to 40 DEG C and rapidly shaken until the cells are completely thawed, and the cells can be directly used without removing the cryopreserving liquid. The method disclosed by the invention can guarantee the quality consistency between batches of cryopreserved cells and the stability of cell genetics, ensure that the survival rate of thawed cells can be stably increased to 98 percent or more, and shorten the cell thawing and passage period by 50 to 60 percent.

Primate embryonic stem cells are cryopreserved by resuspension in a freezing medium and slow cooling at a controlled rate. In some embodiments, prior to the controlled freezing step, the suspension of cells is cooled to a temperature just below freezing, and ice crystal formation is induced. The cryopreserved cell aggregates are useful in transplantation, for experimental evaluation, and as a source of lineage and cell specific products, and as targets for the discovery of factors or molecules that can affect them.

The invention provides a cell cryopreservation agent of hematopoietic stem cells. The cell cryopreservation agent comprises 20 mg of glutamine, 300 [mu]g of epidermal growth factors, 0.2% (w / v) of sucrose, 0.5% (w / v) of low density lipoproteins, 5% (w / v) of trehalose, 0.5% (w / v) of lecithin, 100 ppm (w / v) of polypeptides, 0.6% (w / v) of bFGF, 0.5% (w / v) of vitamin E and 2% (v / v) of glycerol, and a DMEM medium and fetal bovine serum are added to the cell cryopreservation agent according to a ratio of the DMEM medium to the fetal bovine serum of 1:1 until the volume is 100 ml. The glycerol, trehalose, vitamin E and polypeptides in the cryopreserved cells can clearly resist cell injuries brought by outside damages, the polypeptides also protect the cells from being damaged by ice crystals during freezing, and several other components are essential for maintaining the survival of specific hematopoietic cells. The cell cryopreservation agent is used for the cryopreservation of cultured cells, especially improves the survival rate of cryopreserved cells after resuscitation, and has a good application prospect.

The invention discloses a mesenchymal stem cell cryopreservation agent which comprises 0.2%w / v cane sugar, 0.5%w / v low-density lipoprotein, 3%w / v trehalose, 2%w / v lecithin, 100ppm w / v polypeptide, 1.5%w / v bFGF, 1%w / v vitamin E, 5% v / v glycerin and the balance of a DMEM (Dulbecco's Modified Eagle Medium) and fetal calf serum in a ratio of 1:1 till 100ml. In the mesenchymal stem cell cryopreservation agent disclosed by the invention, the glycerin, the trehalose, the vitamin E and the polypeptide have definite functions of preventing external damage to cells, the polypeptide also has a function of preventing cells from damage of ice crystal generated in cryopreservation, and the other components are essential for maintaining survival of specific MSCs (Mesenchymal Stem Cells). The cell cryopreservation agent disclosed by the invention is applied to cryopreservation and culture of cells, is particularly capable of increasing the survival rate of revived cryopreservation cells, and has relatively good application prospects.

The invention discloses a clinical NK cell cryopreservation solution and a cryopreservation method. The clinical NK cell cryopreservation solution comprises the following components in percentage by volume: 3%-10% of dimethyl sulfoxide, 2%-4% of glycerol, 13%-20% of dextran, 0.1%-0.2% of polyvinylpyrrolidone, 5%-10% of N-acetylcysteine and 60%-70% of a glucose solution. Wherein the glucose solution is formed by compounding medical-grade glucose and medical-grade pure water; the mass concentration of glucose in the glucose solution is 5wt%; the clinical NK cell cryopreservation solution can bedirectly used for intravenous injection, the DMSO content is low, the toxicity to human bodies or cryopreserved cells is greatly reduced, and the clinical NK cell cryopreservation solution is safer; the cell viability and killing capability can be better preserved, and the damage to the cells in the cryopreservation process is reduced.