45 results about "Cryopreserved Cell" patented technology

The invention belongs to the technical field of cell cryopreservation, and relates to a cryopreservation method based on ultrasonic ice planting. In the method, the nucleation temperature of ice planting is less than-9 DEG C, and the cooling rate from nucleation to icing is less than 5 DEG C / min. Compared with the prior art, the method has the advantages that the time required for cryopreservation of the cells is remarkably shortened, meanwhile, damage to the cells caused by ice crystals generated in the cooling process is reduced, and the problem that an existing cell cryopreservation cooling method cannot meet the nucleation and freezing uniformity of a cell solution in the cooling process is solved.

Provided are methods for the creation of ultra-high density cryopreserved cell banks. In certain embodiments, these methods employ altered perfusion culture techniques that allow for production of ultra-high density cell cultures that can be cryopreserved at unexpectedly high cell densities without the need for any cell concentration steps, while retaining excellent cell viability and quality.

The invention discloses a novel antifreeze polypeptide with a clear structure and excellent performance and a protection method of cryopreserved cells. The core structure of the polypeptide is formed by covalently connecting two functional modules, the N end is a unit with ice crystal binding performance, the C end is a super-hydrophilic unit, and the two functional modules are bridged through a flexible linker. Experimental verification shows that the synthetic polypeptide shows excellent anti-freezing activity, can significantly inhibit nucleation, growth and recrystallization processes of ice crystals, and has particularly outstanding ice recrystallization inhibition activity. The novel antifreeze polypeptide shows excellent biocompatibility, and is suitable for low-temperature cryopreservation of cells sensitive to freezing damage and other living microbial preparations. Results show that the polypeptide can greatly improve the freeze-drying survival rate of cells and the activity of the cells after recovery, is non-toxic to the cells, and shows the potential of replacing a traditional cryoprotectant. Therefore, the method has wide application prospects in food industry, biological medicine, agricultural breeding and low-temperature preservation technologies of biological sample libraries.

The invention relates to the technical field of cell biology, in particular to an iNKT cell cryopreservation solution and a method for cryopreserving iNKT cells by using the cryopreservation solution. The invention provides an immune cell cryopreservation solution. The immune cell cryopreservation solution comprises human serum albumin, a basic low-temperature protection solution and an optional human platelet lysate. The immune cell freezing medium does not contain animal-derived components except for human bodies, and has the advantages of simple components and good stability. According to the cryopreservation liquid disclosed by the invention, through the combined action of the basic low-temperature protection liquid (such as CryoStor10 (CS10)) and the human serum albumin, the motility rate after cell recovery is improved, and the damage to immune cells (such as iNKT) in the cryopreservation process is reduced. After a small amount of human platelet lysate is further added, the survival rate of cell resuscitation and the amplification capacity can be further improved.

The invention relates to the technical field of biology, in particular to a cryopreserved stem cell diluent and application thereof.The cryopreserved stem cell diluent further comprises cysteine and polyvinylpyrrolidone besides a compound electrolyte injection, the concentration of cysteine in the cryopreserved stem cell diluent is 1-10 mM, and the concentration of polyvinylpyrrolidone in the cryopreserved stem cell diluent is 1-10 mM. The concentration of the polyvinylpyrrolidone in the cryopreserved stem cell diluent is 0.02 to 0.1 g / mL. The cryopreserved stem cell diluent can improve the cell activity and cell distribution after the cryopreserved cell preparation is recovered, and can monitor the application of cells in rat living body imaging.

The utility model provides an anti-pollution cryopreservation tube, and relates to the technical field of cryopreservation tubes. The anti-pollution cryopreservation tube comprises a tube body, the tube body is movably sleeved with a tube cover, a protective sleeve is arranged on the outer wall of the tube body, the outer wall of the protective sleeve is movably sleeved with a protective outer cover, the top face of the protective sleeve makes contact with the bottom face of the tube cover, and the tube cover is located between the protective outer cover and the protective sleeve. The outer wall of the pipe body is sleeved with a floating piece in a sliding mode, and the floating piece is located below the protective sleeve. The sealing performance of the cryopreservation tube can be improved, and water or liquid nitrogen is prevented from permeating into the cryopreservation tube to pollute cryopreserved cells.

Described herein are methods, compositions, and kits for isolation of mitochondria from cryopreserved primary cells (e.g., fibroblast cells, mesenchymal stromal / stem cells) to be used in mitochondrial organelle transplantation (MOT) methods and compositions.

The invention provides a method for quantitatively detecting mitochondrial integrity after cell cryopreservation recovery. The method comprises the following steps: dyeing cells to be detected by adopting a dyeing working solution to obtain dyed cells; the dyeing working solution comprises a JC-10 dyeing working solution or a JC-1 dyeing working solution; mixing the dyed cells with a cryopreservation solution for cryopreservation to obtain cryopreserved cells; wherein the cryopreservation liquid does not contain DMSO (Dimethylsulfoxide) and protein; and after the cryopreserved cells are recovered, directly detecting the mitochondrial integrity of the recovered cells through a dual-emission fluorescence detection method without washing. According to the method for quantitatively detecting the mitochondrial integrity after cell cryopreservation recovery, the cryopreservation liquid which does not contain DMSO and protein is adopted, so that the influence of the cryopreservation liquid on the coloring agent is avoided. The cryopreservation is performed after dyeing, and detection can be directly performed after resuscitation, so that interference caused by mitochondria damage in operation steps such as osmotic pressure change and centrifugation is avoided, and the detection result is more accurate.

The disclosure provides for cryopreservation compositions and methods of use thereof. Aspects of the present disclosure provide for cryopreservation compositions useful in the freezing and thawing of cell therapies. Other aspects of the present disclosure provide for systems for use in the cryopreservation cell therapies. Still other aspects of the present disclosure provide for methods of cryopreserving cell therapies.

The present disclosure provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: dimethyl sulfoxide (DMSO), disaccharide, human serum, and IL-7 and / or IL-15. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w / v % and 10 w / v % dimethyl sulfoxide (DMSO), between about 0.25 w / v % and 5 w / v % disaccharide, and between about 10 w / v % and 90 w / v % human serum. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w / v % and 10 w / v % dimethyl sulfoxide (DMSO), between about 0.25 w / v % and 5 w / v % disaccharide, and between about 0.5 w / v % and 30 w / v % human serum albumin.

The invention discloses an infrared cell resuscitation instrument which comprises a shell and an upper cover movably connected with the shell, an interaction module is arranged on the shell, a control module and one or more resuscitation cavities are arranged in the shell, and an infrared heating module and a temperature sensing module are arranged on the inner wall of each resuscitation cavity. The infrared cell resuscitator is based on an infrared heating principle, does not need to take water as a heat-conducting medium, and can realize efficient cryopreserved cell resuscitation which is not easy to pollute. The infrared cell resuscitation instrument can be further provided with a fan, an optical transmitting unit and an optical receiving unit, so that the cell resuscitation efficiency is improved, and the cell resuscitation effect can be effectively improved.

Described herein are methods, compositions, and kits for isolation of mitochondria from cryopreserved primary cells (e.g., fibroblast cells, mesenchymal stromal / stem cells) to be used in mitochondrial organelle transplantation (MOT) methods and compositions.

The invention relates to a CAR-NK (chimeric antigen receptor-natural killer) cell cryopreservation solution for clinical use as well as a preparation method and application thereof. The CAR-NK cell freezing medium disclosed by the invention is prepared from a washing solution, a dextran 40 glucose injection and Cryostat CS10, or is prepared from the washing solution, human serum albumin and Cryostat CS10; wherein the washing liquid is prepared from sodium chloride injection and human serum albumin. The cryopreservation liquid disclosed by the invention is used for cryopreserving CAR-NK cells, does not have the problem of cell agglomeration, and can effectively maintain the viability and vitality of the cells; the cryopreservation liquid is simple in component, is in a GMP level or a medicine level, can be directly infused back to a human body, is convenient to prepare and easy to prepare in a large scale, and saves the production and manufacturing cost.

The portable self-adaptive multi-specification cryopreserved cell water bath resuscitation device comprises a folding water bath tank and an intelligent buckle cover covering the folding water bath tank, the bottom of the folding water bath is connected with a lining heating film, and the outer periphery of the folding water bath is connected with a plurality of memory metal brackets for folding the folding water bath; the interior of the intelligent buckle cover is connected with a clamping matrix plate layer through a 3D multi-degree-of-freedom rocker arm module. The clamping matrix plate layer is connected with a plurality of sets of hydraulic clamping jaws used for clamping cryopreservation bottles. A distance measuring and temperature measuring module is further connected to the lower portion of the clamping matrix plate layer. The device is convenient to carry, and the 3D multi-degree-of-freedom rocker arm module is adopted to shake a cryopreservation bottle.

The utility model discloses a programmed cooling device for cryopreserved cells. The programmed cooling device is characterized in that an inner container structure is arranged in a box body; a groove is formed in the inner container; the cryopreservation tube frame is arranged in the groove; a plurality of tube grooves for placing cryopreservation tubes are formed in the cryopreservation tube rack; the flip cover is hinged to one side of the box body. According to the cryopreservation tube, the tube sleeve is arranged in each tube groove, so that an independent temperature protection sleeve device is provided for the cryopreservation tube. When the cryopreservation tube is taken out from a low-temperature environment, the tube sleeve can effectively slow down heat exchange between the cryopreservation tube and an external normal-temperature environment, the phenomenon that cells are repeatedly frozen and thawed due to rapid temperature rise is avoided, and the survival rate and activity of the cells are remarkably improved.

The utility model relates to an assembly for taking cryopreserved cells, which aims to solve the technical problems that the current cryopreservation tube is low in temperature and inconvenient to open, and comprises a cryopreservation tube, a connecting ring is integrally formed at the top end of the cryopreservation tube, and a plug is inserted into the top end of the cryopreservation tube; the extraction mechanism is used for extracting the cryopreservation tube, the extraction mechanism comprises tweezers, a connecting groove is formed in the top end of the plug, the fixing pins on the two sides of the tweezers can be inserted into the first fixing grooves in the two sides of the connecting groove in the top end of the plug, and the cryopreservation tube can be driven to be taken out from liquid nitrogen through the tweezers; fixing pins on the two sides of tweezers can be inserted into second fixing grooves in the two sides of a connecting groove, meanwhile, the fixing pins can push out fixing blocks in the second fixing grooves so that the fixing blocks can be contained in limiting grooves, the fixing blocks make contact with fixed connection of the plug and the cryopreservation tube, the tweezers are pulled upwards, and the plug can be pulled out of the top end of the cryopreservation tube; and the cryopreserved cells in the cryopreservation tube can be conveniently and quickly taken by a worker.

The invention discloses a method for accurately determining the motility rate of MSC cells, and belongs to the technical field of motility rate detection of mesenchymal stem cells (MSC). The method mainly comprises the following steps: S1, taking out cryopreserved cells from liquid nitrogen, placing the cryopreserved cells in dry ice, transferring the cryopreserved cells to a laboratory, and carrying out unfreezing resuscitation in a 37 DEG C water bath; s2, taking recovered cells (3E6Cell), averagely transferring the recovered cells (3E6Cell) into three 1.5 ml EP tubes (1E6Cell / tube), adding 1ml of LDPBS, carrying out centrifugation for 5 min, discarding the supernatant, repeating the step once, and then carrying out resuspension by using 100 [mu] l of DPBS for later use; s3, dye preparation: taking one EP tube, adding 400 [mu] l of Annexin V Loading buffer, 10 [mu] l of Annexin V and 10 [mu] l of 7AAD into the EP tube, and carrying out vortex uniform mixing for later use; s4, dyeing: adding 100 microliters of prepared dye into 100 microliters of prepared sample, lightly and uniformly mixing, and incubating in a dark place for 10 minutes; s5, after the dyeing is finished, adding 100 [mu] l of Annexin V loading buffer into each tube, uniformly mixing, and carrying out on-machine detection on a flow cytometer; and S6, taking the average value of the three times of detection data for reporting. The method for accurately determining the cell viability of the MSC achieves the effect of reliable cell counting.

The invention discloses a Super-long fragment DNA extraction kit and application, the Super-long fragment DNA extraction kit is universal, the problem of extraction of DNA super-long fragments of samples such as culture cells, cryopreserved cells, fresh animal tissues, cryopreserved animal tissues, common crops, polysaccharide and polyphenol plants and partial silica gel dry plant samples can be solved, and under the condition that the length reaches 500 kb <-1 > Mb, the extraction rate of the super-long fragments can be greatly improved. The purity can be ensured, and the method is suitable for long-read-length sequencing with high requirements on DNA fragment length and DNA purity.

The utility model relates to the technical field of cryopreservation devices for cryopreservation of cells, in particular to a cryopreservation device for cryopreservation of cells by using liquid betaine, which comprises a liquid betaine storage tank and a freezing tank, and an infusion pump is fixedly connected to one side, far away from the liquid betaine storage tank, of a tank body output pipe. A first injection pipe is fixedly connected to the output end of the infusion pump, a second input port and a third input port are formed in the detachable cover body, a base body is fixedly connected to the bottom end in the freezing tank, a notch is formed in one side of the top of the base body, a freezing cell test tube is arranged on one side in the notch, and a freezing cell is arranged in the freezing cell test tube. A swinging structure fixedly connected with the driving end below the electric telescopic rod is arranged on the other side of the interior of the notch; the frozen cell test tube is driven by the swing structure to move back and forth, so that a cell suspension and liquid betaine in the frozen cell test tube can be mixed, and the trouble of manual shaking is avoided.

The present disclosure provides, among other things, a method of cryopreserving and thawing cells that results in the thawed cells having high cellular viability and functionality post-thawing. In some embodiments, a large-scale method of cryopreserving cells is provided, the method comprising: (a) contacting the cells with a cryopreservation medium; (b) cooling the cells to −80° C. at a controlled rate to minimize latent heat of fusion; and (c) storing the cells in liquid nitrogen vapor phase, thereby cryopreserving the immune cells.

The application discloses a Super-long fragment DNA extraction kit and application. The Super-long fragment DNA extraction kit is universal, can solve the DNA super-long fragment extraction of culture cells, cryopreserved cells, fresh animal tissues, cryopreserved animal tissues, common crops, polysaccharide and polyphenol plants and part of silica gel dry plant samples and the like, can guarantee the purity under the condition that the length reaches 500kb-1Mb, and is suitable for long read length sequencing with high requirements for the DNA fragment length and the DNA purity.

The invention relates to the technical field of cell culture, and discloses a serum-free and DMSO-free human umbilical cord mesenchymal stem cell cryopreservation solution. The invention discloses a serum-free and DMSO (dimethyl sulfoxide)-free human umbilical cord mesenchymal stem cell cryopreservation solution. The serum-free and DMSO-free human umbilical cord mesenchymal stem cell cryopreservation solution is prepared from the following components: ethylene glycol with the final concentration of 1000 to 1500 mmol / L, trehalose with the final concentration of 50 to 150 mmol / L, proline with the final concentration of 50 to 150 mmol / L and 5 to 15 percent of human serum albumin injection. The invention further discloses application of the cell cryopreservation liquid in cryopreservation of cells. The serum-free and DMSO-free human umbilical cord mesenchymal stem cell cryopreservation liquid disclosed by the invention does not contain a dimethyl sulfoxide (DMSO) component and a basic culture medium component, the components are clear, the batch is stable, and the serum-free and DMSO-free human umbilical cord mesenchymal stem cell cryopreservation liquid meets the industrial transformation requirements. Besides, the activity and dryness of the cells can be stably maintained for a long time when the cells are cryopreserved, the recovered cells can be directly used for clinical application, and the safety requirement and the effectiveness requirement of the clinical application are met.

Provided is a method for cryopreserving cells, which is characterized in that the cells are cryopreserved for 30 days or more in a state in which the cells are inoculated on a porous membrane without using liquid nitrogen. Also provided is a cell culture method comprising a step for cryopreserving cells inoculated on the porous membrane for 30 days or more without using liquid nitrogen. Further provided is a cryopreservation base material comprising a porous membrane for cryopreservation of inoculated cells for 30 days or more.

The present invention relates to a frozen cell formulation that can significantly improve the cell viability of cryopreserved cells, preferably stem cells. According to the present invention, the cell viability can be improved when thawing cells after freezing, thereby preventing a decrease in therapeutic efficiency that may occur during the storage and transportation of cell therapy agents.

The present invention relates to a formulation for cell cryopreservation that can significantly improve the cell survival rate of cryopreserved cells, preferably stem cells. According to the present invention, the cell survival rate can be enhanced during the thawing process after freezing, thereby preventing the reduction in therapeutic efficacy that may occur during the storage and transportation of cell therapy products.

The presently disclosed subject matter relates to compositions of ready-to-use cryopreserved populations of dissociated cells which can be directly used for downstream application without after-thaw expansion and / or passage. The presently disclosed subject matter also provides for methods of preparing such compositions, and in vitro methods of using such compositions.

The present disclosure provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: dimethyl sulfoxide (DMSO), disaccharide, human serum, and IL-7 and / or IL-15. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w / v % and 10 w / v % dimethyl sulfoxide (DMSO), between about 0.25 w / v % and 5 w / v % disaccharide, and between about 10 w / v % and 90 w / v % human serum. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w / v % and 10 w / v % dimethyl sulfoxide (DMSO), between about 0.25 w / v % and 5 w / v % disaccharide, and between about 0.5 w / v % and 30 w / v % human serum albumin.

The invention discloses an artificial cell wall system based on a DNA logic gate and application of the artificial cell wall system, the artificial cell wall system comprises a probe 1 (Apt-S-T1) and a probe 2 (T2-S-Apt), Apt is a nucleic acid aptamer targeting a to-be-treated cell membrane receptor, S is a spacer region sequence, and T1 and T2 are a first toe end sequence and a second toe end sequence respectively; a linker; the hairpin comprises a hairpin body 1, a hairpin body 2 and a cross-linking agent, and the tail end of the hairpin body 1 or the tail end of the hairpin body 2 is modified with a hydrogel framework material. Artificial cell walls are constructed based on a DNA logic gate, the integrity of cell membranes is protected in the freezing process, and DMSO-free cryopreservation is achieved; and the resuscitated cell has a biological activation function, so that the resuscitated cell not only can survive, but also has a proliferation capacity superior to that of a fresh cell. Besides, the cell cryopreservation method based on the artificial cell wall system has universality, corresponding artificial cell wall systems can be customized according to different cell types, and cryopreservation requirements of various cell types are met.

The present disclosure provides, among other things, a method of cryopreserving and thawing cells that results in the thawed cells having high cellular viability and functionality post-thawing. In some embodiments, a large-scale method of cryopreserving cells is provided, the method comprising: (a) contacting the cells with a cryopreservation medium; (b) cooling the cells to −80° C. at a controlled rate to minimize latent heat of fusion; and (c) storing the cells in liquid nitrogen vapor phase, thereby cryopreserving the immune cells.

The present invention relates to a preparation for cryopreservation of cells, which can significantly improve the cell viability of cryopreserved cells, preferably stem cells. According to the invention, the cell survival rate can be improved in the thawing process after freezing, so that the possible reduction of the treatment effect of the cell treatment product in the storage and transportation processes is prevented.