36results about How to "Improve cryopreservation effect" patented technology

The invention discloses a direct venous re-transfusion immune cell cryopreservation medium and application thereof. The direct venous re-transfusion immune cell cryopreservation medium is prepared from dimethyl sulfoxide, human serum albumin, dextran, hydroxyethyl starch, heparin sodium and other clinical medicinal level raw materials by optimizing the proportion of the dose of each raw material. In addition, the operation steps are simplified and the cryopreservation effect is improved by optimizing the cryopreservation method. On one hand, the potential risk of introducing an animal source antigenic substance in the cell treatment process is avoided, and on the other hand, the cryopreservation effect of immune cells is better than that of a conventional cryopreservation medium, thereby guaranteeing the safety and validity in clinical use. Two kinds of components are used for preservation, so that the cryopreservation medium can be preserved stably for a long time. The method disclosed by the invention is simple in operation, provides stronger operability for implementation of large-scale cryopreservation, and simultaneously provides a fundamental technical guarantee for implementation of immune cell treatment.

The invention discloses new application of tetrahydropyridines, namely the application of the tetrahydropyridines to the cryopreservation of cells, tissues and organs. The effective concentration is 0.1g / L to 100g / L; and the preferred concentration is 0.5g / L to 20g / L. During use, the tetrahydropyridines can be used separately or can be jointly used with other chemical reagents for the cryopreservation of cells, tissues and organs. Through experiments, the inventor of the application verifies that: the tetrahydropyridines can help the cells in resisting the refrigeration conditions and improve the protective effect on the cryopreservation of cells, tissues and organs. If the tetrahydropyridines is jointly used with dimethyl sulfoxide, the tetrahydropyridines can reduce the dosage of the dimethyl sulfoxide, reduce the toxicity of the dimethyl sulfoxide on the cells, and improve the cryopreservation effect.

The invention belongs to the field of cell biology and in particular relates to a cryopreservation solution and a cryopreservation method of a DC cell. The cryopreservation solution of the DC cell contains trehalose, bovine serum albumin, propylene glycol, DMSO, low molecular dextran, a DMEM and vitamin C. The cryopreservation method of the DC cell is characterized by adding the cryopreservation solution to the DC cell, mixing the cryopreservation solution with the DC cell uniformly and then cryopreserving the mixture. The cryopreservation solution and the cryopreservation method have the beneficial effects that the cryopreservation effects of the DC cell are obviously improved by adopting the cryopreservation solution to cryopreserve the DC cell, so the DC cell can maintain a better state in either the viability after resuscitation or proliferation.

The invention provides a method for rapidly cryopreserving and resuscitating endometrial stem cells. The method is capable of reducing the probability of cell damage and apoptosis and improving the survival rate. According to the method, cryopreservation fluid or resuscitation fluid is added into the cells to perform short-term low-frequency ultrasonic treatment, so that mutually adhesive cells can produce slight loosening and gaps on premise of not damaging the cell structure, and the fluid can be dispersed into droplets and conveniently infiltrates into the cells; cell adhesion molecules arecapable of promoting mutual adhesion of the cells, so that individual cells are further reduced, and the cell survival rate is improved; and a cell activity inhibitor is capable of performing reversible inhibition on some physiological activities in the cells, and the cryopreservation effect is improved on premise of not influencing the cell activity. According to the means, physical and chemicaldamage to the cells in the cryopreservation and resuscitation process can be reduced, so that the cell survival rate and cryopreservation and resuscitation rate of the cells are improved.

The invention belongs to the technical field of biology and particularly relates to protein-free liquid-nitrogen cell frozen preservation liquid and a preparation method thereof. Each 100ml of protein-free liquid-nitrogen cell frozen preservation liquid contains 2.0-15.0g of any one of dextran, saccharosan and hydroxyethyl starch, 0.2-2.0g of poloxamer 188, 10ml of DMSO (Dimethylsulfoxide) and the balance of an RPMI-1640 (Roswell Park Memorial Institute-1640) cell basic culture medium. The protein-free liquid-nitrogen cell frozen preservation liquid provided by the invention is prepared from chemical components and does not contain protein; and the recovery rate of cells is high and the stability is good.

The invention provides an umbilical cord tissue freeze preservation and unfreezing method. According to the umbilical cord tissue freeze preservation and unfreezing method, cell aggregate freeze preservation is adopted instead of single cell freeze preservation, so that cell damage apoptosis rate is reduced, and survival rate is increased; cell adhesion molecules are added to promote bonding of cells, reduce the amount of single cells, and increase cell survival rate; ultrasonic treatment is adopted so as to ensure full penetration of a refrigeration protective solution and an unfreezing protective solution into intercellular spaces, solutions are dispersed into small liquid drops, and the permeability is improved; a cell activity inhibitor is adopted for reversible inhibition of physiological activities in cells, and improving of freeze preservation effect without influencing cell activity. The umbilical cord tissue freeze preservation and unfreezing method is capable of reducing physical and chemical damages on cells in freezing and unfreezing process, and increasing cell survival rate and unfreezing rate, so it is convenient for subsequent using.

The invention relates to a method using umbilical cord tissue cryopreservation to preserve various stem cells, a thawing method and application. The method specifically includes: cleaning umbilical cord tissue, shearing into tissue sections, and stripping mesenchyme Wharton's jelly tissue; using an alpha-MEM protecting agent containing DMSO for preprocessing, performing concentration balance under4-10 DEG C for 10-20 minutes, performing low-temperature centrifuging, and removing supernate; adding dextran or fetal calf serum to allow the dextran or fetal calf serum and the alpha-MEM protectingagent containing DMSO to jointly form cryopreservation liquid, and using a program-controlled temperature lower or graded temperature lowering manner to perform cryopreservation on the tissue. The cryopreservation method has the advantages that the cryopreserved umbilical cord tissue can be effectively protected, subsequent thawing operation is facilitated, and the activity of cells cultured by the thawed tissue is the same as that of cells in fresh tissue.

The invention discloses a boar seman cryoprotectant which is prepared from the following substances: cryopreservation basic liquid, fresh egg yolk, glycerinum, astaxanthin and a radix rehmanniae preparata extract. The boar seman cryoprotectant is subjected to special matching treatment; under the common action of all the components, the finally prepared cryoprotectant obviously enhances the boar seman cryopreservation effect, and increases the boar seman viability, acrosomal integrity and film integrity rate; the fertilization vitality and the use effect of boar semans are guaranteed to the maximum extent.

The invention discloses a predicting method of best cooling rate of red blood cell in cryopreservation state, comprising the steps as follows: according to the linear relationship between the heat release of the cell suspension solution during cooling process and the cell volume, ensuring the volume change of the cell in a specific storage solution under constant cooling rate; according to the mathematical model of water transmembrane transport, fitting a percolation parameter corresponding to the red blood cell, simulating the volume changes of the cell under different cooling rates to predict the best cooling rate scope. The cell recovery demonstration tested by the cell cytometer shows that the DSC method can correctly predict the best cooling rate of the cell in the specific storage solution and can greatly reduce the work intensity and is convenient to guide and create a more reasonable low-temperature storing solution.

The invention relates to the technical field of cryopreservation solutions, in particular to a cell cryopreservation solution, a method for cryopreserving hematopoietic stem cells by using the cell cryopreservation solution and a stem cell preparation. The invention discloses a cell cryopreservation solution which is composed of DMSO, a human serum albumin injection and a plasmon A injection. Thecell cryopreservation liquid is simple and safe in components, the components are matched with one another, a synergistic effect is achieved, the cell cryopreservation effect can be remarkably improved, the survival rate of cryopreserved hematopoietic stem cells after recovery is high, the cell stability is good, and treatment of diseases is facilitated.

The invention relates to an ultrasonic plant freezing and storing device, in particular to an ultrasonic plant freezing and storing device for hepatocytes and a freezing and storing method of the ultrasonic plant freezing and storing device. The ultrasonic plant-freezing storage device comprises an ultrasonic container, a temperature detection system, an ultrasonic induced nucleation system and a cooling system, the liver cell ultrasonic plant-freezing preservation method comprises the following steps: culturing liver cells, determining an upper threshold of a plant-freezing temperature, selecting a temperature-controlled phase-change liquid raw material, and carrying out liver cell ultrasonic plant-freezing preservation. Compared with a conventional hepatocyte cryopreservation mode, the ultrasonic plant-freezing cryopreservation device and the ultrasonic plant-freezing cryopreservation method have the advantages that the hepatocyte cryopreservation effect is remarkably improved, the use amount of the cryoprotectant is small, the ultrasonic plant ice and the cryopreservation protectant have a synergistic effect, the recovery survival rate of the cryopreserved hepatocytes is higher than 90%, and no significant difference exists between the cryopreserved hepatocytes and fresh groups; and an unexpected technical effect is achieved.

The invention provides an immune cell cryopreservation solution and an immune cell cryopreservation method, which are characterized in that the immune cell cryopreservation solution is composed of twoindependent cryopreservation solutions, namely, a cryopreservation solution A and a cryopreservation solution B, and the two cryopreservation solutions are mixed for use according to a ratio of 1: 1(v / v); and when immune cells are cryopreserved by using the immune cell cryopreservation solution, the immune cells are re-suspended first by using the cryopreservation solution A, the cryopreservation solution B with the same volume is then added, the mixed solution is uniformly blown and beaten and sub-packaged into cryopreservation containers, and the cryopreservation containers are transferredinto an ultralow-temperature refrigerator and then transferred into liquid nitrogen for storage after 24 hours. Compared with the existing cell cryopreservation technology, the immune cell cryopreservation solution and the immune cell cryopreservation method have the advantages that immune cells are effectively protected, the operation is simple, the cryopreservation effect is good, the immune cells are not damaged in the cryopreservation process, the safety is high, the physiological function and biological characteristics of the recovered immune cells are guaranteed, the completeness of surface antigens of the immune cells is guaranteed, and the survival time of the immune cells is prolonged.

The invention provides a stem cell cryopreservation liquid and a stem cell cryopreservation method, and the stem cell cryopreservation liquid comprises the following components by volume percentage: a serum-free GT-T551H3 culture medium containing 50%-60% of an alpha-MEM basic culture medium; 15%-20% of human serum albumin with a concentration of 15-35 g / l; 1% to 5% of sulfosalicylic acid; 5% to 10% of acetamide; 8%-12% of dimethyl sulfoxide; wherein the volume ratio of sulfosalicylic acid to acetamide is (1: 1)-(1: 10). The stem cell cryopreservation liquid provided by the invention has the advantages; the cell cryopreservation liquid does not contain animal serum, so that the risk of introducing pollution and allergens can be avoided, and the cell cryopreservation liquid has higher clinical safety compared with a conventional cell cryopreservation liquid; and the sulfosalicylic acid and acetamide components can prevent the cells from being damaged due to formation of ice crystals in the cryopreservation process.

The invention discloses a freezing preservation solution used for adipose-derived stem cells as well as a preparation method for the freezing preservation solution and a culture medium. The culture medium comprises the following components in parts by weight: 7200-7300 parts of inorganic salt composite, 980-1000 parts of amino acid complex, 40-45 parts of vitamin composite, 1000-1100 parts of glucose, 100-110 parts of sodium pyruvate, 60-75 parts of butyric acid, 80-110 parts of sodium butyrate, 0.15-0.22 part of ferric citrate and 1,000,000 parts of water. The freezing preservation solution has the beneficial effects that: the culture medium has excellent freezing preservation effect in the freezing preservation solution which is compounded by a serum substitute component KSR and dimethyl sulfoxide, and is used for the adipose-derived stem cells.

The invention relates to a swine semen freezing protecting agent and a cryopreserving method for swine semen by using the same. The swine semen freezing protecting agent is prepared from the followingcomponents in parts by weight: 2.5 parts of glucose, 2 parts of citric acid, 3 parts of coconut oil monoethanolamide, 2.5 parts of hyaluronic acid, 1.5 parts of skimmed milk, 1.2 parts of trehalose,0.07 part of penicillin sodium, 0.2 part of streptomycin sulfate, 95 parts of double distilled water, 23 parts of fresh yolk, 3.5 parts of glycerin, 1.5 parts of astaxanthin and 5 parts of extract ofradix rehmanniae praeparata. The cryopreserving method comprises the following steps of firstly, collecting the swine semen, centrifuging, and filtering, so as to obtain the swine semen A; pouring onepart of swine semen A into an experiment bottle, and adding two parts of freezing protecting agent to thin; finally, adding into liquid nitrogen, and cryopreserving. The swine semen freezing protecting agent has the advantages that the freezing protecting agent is reasonably compounded, so that the cryopreserving effect of the swine semen is improved under the function of the components; the survival rate, acrosome integrity and plasmalemma integrity of the swine semen are improved; the insemination activity and use effect of the swine semen are guaranteed.

The invention relates to the technical field of embryo transplanting and in particular relates to an embryo production and refrigeration technology, aiming at improving the refrigeration and preservation effect of embryos and improving the activity of the embryos when the embryos are unfrozen and transplanted. The embryo production and refrigeration technology comprises the following steps: (1) selecting a donor ewe and a donor ram; (2) collecting sperm from the donor ram; (3) enabling the donor ewe to superovulate; (4) inseminating; (5) collecting the embryos; (6) identifying the embryos; (7)preparing a solution needed by refrigeration; (8) pre-treating the embryos before refrigeration; (9) filling the embryos into tubes; (10) refrigerating; (11) preserving.

The invention discloses a sheep seminal fluid cryoprotectant and an application thereof, wherein the sheep seminal fluid cryoprotectant comprises a freezing base diluent and an AMPK activator, and the AMPK activator is Met or AICAR. By adding the AMPK activator Met or AICAR with a proper concentration into the sheep seminal fluid cryopreservation diluent, the vitality, movement performance and structural integrity of unfrozen sperms can be improved by activating AMPK, and the seminal fluid quality is improved by maintaining acrosin activity, improving antioxidant enzyme activity, promoting sperm metabolism and maintaining mitochondrial functions.

The invention relates to macrophage cryopreservation liquid and a macrophage cryopreservation method. The macrophage cryopreservation liquid is prepared from the following components: glucan, bovine serum albumin, sorbitol, low molecular dextran, chitosan and RPMI 1640. The macrophage cryopreservation method utilizes the macrophage cryopreservation liquid to cryopreserve macrophages. According to the macrophage cryopreservation liquid and the macrophage cryopreservation method, the cryopreservation effect of the macrophages is improved, the survival rate is relatively high after recovery, the growth state of cells is good and the generation time is relatively long; the macrophage cryopreservation liquid does not utilize FBS (Fasting Blood Sugar) so that the safety of clinical transfusion is greatly improved; and the content of DMSO (Dimethylsulfoxide) is reduced and the toxin risks to human bodies are reduced.

The invention discloses a cryopreservation method for pinus elliottii embryogenic calluses resistant to pine needle spot brown spot. According to the method provided by the invention, proper pre-culture and a mixed cryoprotectant facilitate improving the cryopreservation effect, after thawing is performed at 32 DEG C, the calluses frozen for 90 days can be regenerated after 1 month, the regeneration rate of the embryogenic calluses is up to 100%, and the regenerated calluses grow well and still retain the ability to differentiate mature somatic embryos; and the method preliminarily establishesa cryopreservation system for the pinus elliottii embryogenic calluses resistant to the pine needle brown spot, and provides a technical reference for embryogenic long-term maintenance of the resistant pinus elliottii embryogenic calluses.

The invention discloses a cell seedling temperature cryopreservation method and a device, and belongs to the technical field of cell cryopreservation, the method comprises the following steps: S1, cooling a cryopreservation tube loaded with cells to be cryopreserved through a refrigeration system according to a preset cooling rate, and detecting the temperature of the cryopreservation tube in the cooling process; S2, judging whether the temperature of the cryopreservation tube reaches a preset seedling temperature or not, if yes, executing S3, and if not, executing S1 continuesly; S3, adopting the refrigeration system to conduct heat preservation on the cryopreservation tube at the preset seedling temperature; S4, applying ultrasonic waves to the cryopreservation tube through an ultrasonic vibrator so as to carry out seedling temperature operation, and lasting the operation for preset threshold time; S5, judging whether seedling temperature operation of the cryopreservation tube succeeds or not by analyzing the temperature of the cryopreservation tube before and after the seedling temperature operation, if yes, executing S6, and if not, continuing to execute S4; and S6, cooling the cryopreservation tube continuesly through the refrigerating system, stopping cooling after the cryopreservation tube is cooled to the preset cryopreservation temperature, and carrying out heat preservation. According to the method, the cell cryopreservation effect is improved by adding the step of judging whether seedling temperature operation succeeds or not.

The invention relates to the field of biotechnology, in particular to a cord blood mononuclear cell (CBMC) cryopreservation solution and application thereof. The CBMC cryopreservation solution is prepared from dimethyl sulfoxide (DMSO) combined hydroxyethyl starch (HES), propylene glycol, human albumin and a buffer solution. Compared with a conventional formula, the CBMC cryopreservation solutioncan avoid contamination of animal-derived pathogens, does not take a cell culture medium as a component of a cryoprotectant, and ensures good cell cryopreservation effect. After cryopreservation for 2months, the average cell viability of CBMC cryopreserved with the cryopreservation solution is 97.47% to 98.43%. The recovered cell can be directly used for clinical transplantation, and the CBMC cryopreservation solution is of practical value in clinical application of cell therapy.

The invention relates to the technical field of stem cell culture, in particular to a protein-free non-programmed freezing medium for umbilical cord mesenchymal stem cells as well as a preparation method and application of the protein-free non-programmed freezing medium. The cryopreservation solution comprises a basic solution, a nutritional supplement, a permeable protective agent, a non-permeable protective agent, a cell sedimentation stabilizer, a cell membrane protective agent, an apoptosis inhibitor and an antioxidant, the basic solution is DMEM / F-12 with the glucose content being smaller than or equal to 1000 mg / L. The cryopreservation liquid is suitable for directly cryopreserving umbilical cord mesenchymal stem cells at-80 DEG C after in-vitro amplification and before stem cell treatment. The cryopreservation liquid is free of serum and protein and clear in chemical component, and the cryopreserved umbilical cord mesenchymal stem cells are free of exogenous pollution risk and safer to use; after the cells are recovered, the viability is high, the adherence rate is high, and the cell expansion is fast; surface marker characteristics (phenotypes) and three-line differentiation potential of the mesenchymal stem cells can be maintained; optimization is carried out according to the cryopreservation and culture characteristics of the umbilical cord mesenchymal stem cells, the cryopreservation effect is improved, programmed cooling is not needed, and time and labor are saved.

The invention provides a method for improving an encapsulation-dehydration ultralow-temperature preservation effect of early pear stem tips. The early pear stem tips are treated in sequence by utilizing four types of culture media containing carbon nanotubes and graphene quantum dots, and the survival rate of the frozen stem tips can be remarkably improved. Compared with the prior art, the methodhas the advantages that the encapsulation-dehydration ultralow-temperature preservation effect of the early pear stem tips is good, the survival rate is high and the stem tips are easy to regenerate.

The invention provides an improved mesenchymal stem cell protective solution. The concentration of electrolytes in an unfrozen water solution inside and outside cells is balanced through a permeatingprotective agent, so that cell damage due to excessive dehydration of cells in the outside high-permeability environment is avoided; added protein composition can supply cells with required nutrients,and accordingly, the cell survival rate is increased; a physiological activity inhibitor of cells can reversibly inhibit physiological activities in the cells, and the cryopreservation effect is improved on the premise that the cell activity is not affected; amifostine is a broad-spectrum cell protective agent and a radiation protective agent and can improve the tolerance of cells to the outsideadverse environment, reduce accumulation of toxic substances in cells and improve the proliferative activity of stem cells. Through combined use of components, the stress resistance of mesenchymal stem cells can be effectively improved, cell activity loss caused in the cryopreservation process can be reduced, and the cell survival rate and the thawing rate after cryopreservation are increased.

The invention discloses a method for cryopreservation of embryogenic callus of pine needle brown spot disease-resistant slash pine. Appropriate pre-cultivation and the use of mixed cryoprotectants can help improve the effect of cryopreservation. After thawing at 32°C, the callus frozen for 90 days can resume regeneration after 1 month, and the regeneration rate of embryogenic callus is the highest. 100%, the regenerated callus grows well and still maintains the ability to differentiate into mature somatic embryos. This application preliminarily established a cryopreservation system for embryogenic callus of pine needle brown spot disease-resistant wetland pine embryogenic callus, providing technical reference for long-term maintenance of embryogenic callus of resistant wetland pine callus.

The invention provides a method for improving a droplet-vitrification ultralow-temperature preservation effect of early pear stem tips. The early pear stem tips are treated in sequence by utilizing six types of culture media containing carbon nanotubes and graphene quantum dots, and the survival rate of the frozen stem tips can be remarkably improved. Compared with the prior art, the method has the advantages that the droplet-vitrification ultralow-temperature preservation effect of the early pear stem tips is good, the survival rate is high and the stem tips are easy to regenerate.