49 results about "Platelet lysate" patented technology

The invention relates to the technical field of hydrogel, in particular to composite hydrogel capable of stopping bleeding, resisting bacteria and promoting alveolar bone repair, a preparation method and application. The composition comprises a polyacrylic acid solution, a calcium chloride solution, a disodium hydrogen phosphate solution, platelet-rich plasma (including plasma, platelet lysis buffer, platelet-rich fibrin and fibrin of high-concentration concentrated growth factors) and nano clay (hectorite). The preparation method has the advantages of simple process, easily available raw materials, environment-friendly production process, safety and short preparation period. The composite hydrogel is good in biocompatibility and injectable, and can be widely applied to the field of biomedical materials.

Platelet lysate compositions and cell culture media compositions for maintaining and / or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and / or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and / or flasks pretreated with the cell culture media compositions.

A sterilised platelet lysate obtained with a method for sterilising a platelet lysate in the liquid state comprising at least the endogenous growth factors TGF-beta 1, EGF, PDGF-AB, IGF-1, VEGF and bFGF. The method comprising freezing the liquid platelet lysate in order to obtain a frozen platelet lysate, and irradiating the frozen platelet lysate with ionising radiation in order to obtain a sterilised platelet lysate, the irradiation being adapted so as to preserve at least 80% of the concentration of at least one of the endogenous growth factors chosen from the group consisting of TGF-beta 1, EGF, PDGF-AB, IGF-1 and VEGF.

The invention belongs to the field of medicine health. Specifically, the invention provides preparation and application of a lysate of platelets differentiated in vitro. Specifically, the invention provides application of the lysate in effectively promoting migration of cells to a skin injury, reducing inflammation, limiting matrix degradation and promoting collagen synthesis to support wound healing.

The present invention relates to, inter alia, various scaffold materials including a scaffold material comprising: i) platelet-rich plasma and / or platelet lysate and / or bone marrow concentrate; and ii) electrospun fibres comprising silk protein.

The invention discloses an NK cell amplification culture solution and application thereof, the amplification culture solution comprises a basic culture medium, and the basic culture medium comprises polysaccharide with the final concentration of 1-500 [mu] g / mL, IL-2 with the final concentration of 50-2000 U / mL, platelet lysate with the final concentration of 0.5-5% v / v, a concentrated extract of macaque rhizomes with the final concentration of 0.1-1% v / v and a concentrated extract of gentian rhizomes with the final concentration of 0.1-1% v / v. According to the NK cell amplification culture solution, the concentrated extract components of the macaque rhizomes and the gentian rhizomes can maintain the activity of NK cells, slow down cell aging, promote secretion of cell factors and maintain long-period (24 days or more) cell proliferation; meanwhile, the NK cell culture process is simple, coating is not needed, and operation is convenient.

The application is a preparation of a composite stem cell active biomaterial and its application in artificial skin, effectively solves the problems of high production cost, complex production process and non-use for active substance research of the existing method, and comprises a double-layer composite material, the lower layer is an active material layer in the form of hydrogel composed of active liposomes and stem cells, and the upper layer is an antibacterial layer; the preparation method of the thiolated hyaluronic acid / H active factor composite hydrogel is relatively mild, does not need to add a crosslinking agent and contains a large amount of active ingredients; by using the mechanism, the environment can induce angiogenesis through endothelial cells / fat stem cells combined with active factors and platelet lysate and rapidly construct microtissues in vitro, which is beneficial to the scaffold in large-scale cultivation of artificial skin basement layer in vitro to meet the needs of patients.

The purpose of the present invention is to provide a method for preparing a dendritic cell from a monocyte using a platelet lysate.　Provided is a method for preparing a dendritic cell having cytotoxicity from a monocyte, the method comprising culturing a monocyte separated from peripheral blood by non-adhesive culture using a serum-free culture medium containing a human platelet lysate (HPL), GM-CSF and PEG conjugated interferon-α, adding prostaglandin E2 and OK432 to the resultant culture, and further culturing the resultant mixture by non-adhesive culture.

The application discloses a method for inducing T cell reprogramming into NK-like cells, comprising the following steps: (1) culturing the T cells in a culture system to obtain activated T cells; (2) adding small molecule inducers twice to the activated T cells to obtain NK-like cells; wherein the culture system in step (1) contains 1-10 v / v % serum substitute and / or platelet lysate, and does not contain fetal bovine serum. By optimizing the culture process and precisely adding drugs twice, the small molecule accumulation toxicity caused by multiple drug additions is avoided, the damage to the structure and function of the mitochondrial membrane is reduced, the mitochondrial membrane potential is significantly improved, the energy metabolism capacity of the mitochondria is enhanced, sufficient energy support is provided for the long-term survival and function maintenance of the cITNK cells, and the physiological activity of the cells is ensured from the energy metabolism level.

The present disclosure discloses a method for isolating osteoprogenitors like mesenchymal stem cells (MSCs) from clotted bone marrow and culturing with a platelet lysate obtained from a combination of discarded umbilical cord blood and maternal blood platelet-rich plasma (instead of non-human animal origin serum) and differentiating those MSCs into osteoblasts under sterile conditions for further therapeutic applications. Particularly, the present disclosure relates to a method for expansion of osteoblasts to make cell therapy products with a fixed cell dose, which are characterized and later cryopreserved for future use through its cell culture process. Further, the present disclosure relates to identifying specific gene expression from MSCs to osteoblast formation, an in-vitro differentiation process that replicates the in-vivo bone remodelling system.

The application discloses a composition for inducing T cell reprogramming into NK-like cells and application thereof. The composition comprises a DNA methyltransferase inhibitor, a histone deacetylase inhibitor and a vitamin C phosphate derivative; the molar ratio of the DNA methyltransferase inhibitor and the vitamin C phosphate derivative is 1: (40-2500); and the final concentration of the histone deacetylase inhibitor is 0.05-2 muM. The vitamin C phosphate derivative is used to replace common vitamin C, and the defects of common vitamin C, such as weak reprogramming promotion effect in a platelet lysate system, limited cell proliferation and low receptor expression, are overcome, so that the composition is more suitable for a clinical serum-free large-scale preparation scene.

Disclosed are: a plasma solution containing a plasma coming from a first leukocyte removal filter through which human whole blood has been passed and a plasma coming from a second leukocyte removal filter through which the human whole blood has been passed, wherein the plasma coming from the second leukocyte removal filter is a plasma contained in a solution obtained by passing, through the second leukocyte removal filter, a solution containing the plasma coming from the first leukocyte removal filter; a platelet lysate; a method for producing the plasma solution; and a method for producing the platelet lysate.

The present invention relates to, inter alia, various scaffold materials including a scaffold material comprising: i) platelet-rich plasma and / or platelet lysate and / or bone marrow concentrate; and ii) electrospun fibres comprising silk protein.

The invention relates to methods for extracting, purifying and concentrating mesenchymal stromal cell exosomes enriched with micro-RNA, proteins and lipids, and can be used in cosmetology and pharmacology for rehabilitation and regeneration. The technical result of the invention is a more efficient process for producing exosomes. This technical result is achieved in that the present method for extracting, purifying and concentrating exosomes from mesenchymal stromal cells includes cultivating cells, collecting the conditioned culture medium, depleting high-molecular-weight proteins, extracting, purifying and concentrating exosomes, and freezing and lyophilizing the exosome concentrate; in order to enrich the culture medium with exosomes, multipotent mesenchymal stem cells (MSCs) are grown in a medium having a lowered glucose concentration and containing platelet lysate and 5 mM of L-alanyl-L-glutamine, wherein, in the case of static cell cultivation, the growth medium is placed in a culture vessel and once a level of not less than 80% monolayer confluence is reached, the culture medium is harvested and replaced once every 24 hours, and in the case of flow-through cell cultivation, cells are placed on cell carriers in a bioreactor and 72-96 hours after the start of cultivation, the culture medium is harvested and replaced once every 24 hours, the exosome-enriched culture medium is subjected to concentration, which includes successive stages of centrifugation, tangential flow filtration and isopycnic centrifugation: in the first stage, the culture medium is centrifuged for 10 mins at 2000 g to remove large cell particles and other inclusions; in the tangential flow filtration stage, the supernatant is concentrated using hollow fibre filters having a molecular weight cutoff of 500 kDa, after which the concentrate is subjected to fivefold diafiltration with an equal volume of DPBS phosphate buffer solution using the same hollow fibre cartridge; and in the isopycnic centrifugation stage, the concentrated supernatant is mixed with a saccharose solution and deuterium oxide and subjected to isopycnic centrifugation at 100000 g and 4˚С for 75 mins, after which the concentrate is subjected to further fivefold diafiltration with a buffer solution, then the concentrate is filtered through filters with a pore diameter of 0.22 µm and 0.1 µm, the exosome concentrate is subjected to initial freezing to -75˚C in a freezing chamber, and drying is carried out in a lyophilization chamber, wherein in a first drying stage, the concentrate is cooled to -40˚С and subsequently held for 100 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -30˚С and subsequently held for 610 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -10˚С and subsequently held for 315 mins at an absolute pressure in the chamber of not more than 10 Pa, after which it is cooled to 0˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa, then heated to +10˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa; in a second drying stage, drying is carried out at +22˚С for not less than 130 minutes at an absolute pressure in the chamber of not more than 10 Pa.

The application discloses a mesenchymal stem cell large-multiple expansion scheme and belongs to the technical field of biology. The method uses cryopreserved umbilical cord mesenchymal stem cells as seeds (including P1 / P2 / P3 / P4 / P5), rapidly recovers the seed cells, uses a mixture of platelet lysate and serum substitute as a culture medium additive to carry out culture, and achieves the method of large-multiple expansion of mesenchymal stem cells. Compared with a traditional single culture medium culture mode, the method has the advantages of different culture systems, can achieve large-multiple expansion of cells, improves production efficiency, has low cell damage, has stable and uniform cell diameters, has slow aging speed of cell growth, can still keep the cell state and the cell survival rate at a high level after multiple passage expansion, and can avoid the problems of insufficient quantity of mesenchymal stem cells in a single batch and uneven quality between multiple batches, and thus influences clinical application.

The present invention provides a method for preparing a platelet lysate rich in extracellular vesicles and its application. The platelet lysate prepared by intermittent ultrasonic lysis does not require the exogenous addition of biochemical components such as thrombin, collagen, or calcium ions, making it simpler and safer. By setting reasonable ultrasonic process parameters, the prepared platelet lysate is not only rich in various factors (such as bFGF, VEGF, PDGF-BB, and IL-6, IL-15, IL-17, and INF-γ) but also has a high content of bioactive proteins. Furthermore, the extracellular vesicles are not only high in content but also small in diameter, with an average diameter of as low as 79.9 nm. The proportion of exosomes is relatively large, and the extracellular vesicles have high expression of the transmembrane protein CD9 and low expression of CD81.

A method for proliferating Wharton's jelly mesenchymal stem cells comprises the steps of (a) isolating Wharton's jelly tissues comprising viable Wharton's jelly mesenchymal stem cells; (b) partially digesting the isolated tissues by using first culture medium comprising collagenase type I and antibiotic-antimycotic; (c) preparing a primary culture of Wharton's jelly mesenchymal stem cells in suspension culture by using a second culture medium comprising antibiotic-antimycotic and platelet lysate; (d) expanding the primary culture in the suspension culture; (e) obtaining a subculture from the expanded primary culture; and (f) harvesting viable Wharton's jelly mesenchymal stem cells from the subculture.

The present disclosure relates to a process to assemble proteins derived from platelet lysates in bioactive microparticles, with increased surface organization. The present invention further relates to protein-based biomaterials applicable to biomedical and biotechnology fields, more precisely in tissue engineering strategies, disease modeling, and other biomedical applications. Namely, a method for obtaining a protein microparticle from a platelet lysate comprising the following steps: lyophilizing the platelet lysate; dissolving the lyophilized platelet lysate in phosphate buffer saline to obtain a platelet lysate solution; adding a reducing agent to the platelet lysate solution to obtain a reduced platelet lysate solution; adding an oxidizing agent to the reduced platelet lysate solution to form the microparticles by precipitation.

Methods provided for producing human platelet lysates (HPL) typically from expired platelet units that were initially manufactured to be infused into patients. Whole blood units from which platelets will be prepared for transfusion are maintained at a temperature cooling toward room temperature. The platelet lysate produced by the method can be formed from platelets that have been removed from the refrigerated whole blood unit more than 8 hours after phlebotomy.

The invention provides a non-heterogeneity serum-free culture medium for large-scale culture of human umbilical cord mesenchymal stem cells and a culture method. Specifically, the present invention provides a serum-free cell culture medium comprising a basic culture medium and additive components wherein the additive components comprise transferrin, insulin, selenium or a salt thereof, transforming growth factor-beta (TGF-beta), a platelet lysate composition, and a glutamine supplement. The invention further provides a method for culturing the mesenchymal stem cells through the culture medium and application of the mesenchymal stem cells. The culture medium disclosed by the invention has an excellent supporting effect on cell morphology, cell proliferation capacity, multiplication time and the like.

The invention discloses an in-vitro amplification method of bone marrow mesenchymal stem cells, which comprises the following steps: taking DMEM (Dulbecco Modified Eagle Medium) as a basic culture medium, and adding platelet lysate (PL), L-glutamine, epidermal growth factor (EGF), fibroblast growth factor (FGF) and endothelial growth factor (VEGF); inoculating the separated and purified bone marrow mesenchymal stem cells on a three-dimensional porous scaffold, and placing the three-dimensional porous scaffold in a dynamic culture system; periodic mechanical stimulation is applied under the low-oxygen condition; and replacing the culture medium at set intervals, and carrying out periodic culture until the cell density reaches 1 * 10 < 6 >-5 * 10 < 6 > cells / mL. The invention relates to the technical field of stem cell culture. According to the in-vitro amplification method for the bone marrow mesenchymal stem cells, a culture medium formula is optimized, a three-dimensional dynamic culture system is combined, the cell amplification efficiency and quality are improved, and powerful support is provided for stem cell treatment.

A particulate lyophilized platelet lysate composition suitable for use as a cell culture medium can include growth factors, cytokines, and chemokines released from lysis of source platelets, wherein cellular debris from the source platelets is removed (partially or fully) by filtration. The growth factors, cytokines, and chemokines are lyophilized to form a particulate lyophilized platelet lysate composition.

The application provides a serum-free cell cryopreservation solution for epidermal stem cells and a preparation method and application thereof, and specifically belongs to the technical field of stem cell cryopreservation.The serum-free cell cryopreservation solution for epidermal stem cells comprises the following components in the following amounts: 0 g / L to 50 g / L raffinose, 3 g / L to 30 g / L dextran, 0% to 30% (volume percent) glycerol, 0% to 10% (volume percent) DMSO, 0% to 30% (volume percent) platelet lysate, and the balance of Ham's F-12; wherein the values of DMSO and glycerol are not both 0%. The serum-free cell cryopreservation solution can significantly improve the proliferation activity and colony formation ability of epidermal stem cells after resuscitation.

The invention relates to the technical field of polymer glycosylation analysis, and discloses a VWF polymer glycosylation detection method based on gel protein electrophoresis, and the method comprises the following steps: carrying out denaturation treatment on VWF-rich plasma, platelet lysis buffer or cell supernatant to prepare a sample; the method comprises the following steps: carrying out low-temperature constant-pressure electrophoresis on a sample by adopting agarose gel protein electrophoresis, separating VWF polymers with different glycosylation degrees, transferring colloids after electrophoresis to a hydrophilic elastic polyester film, and carrying out air drying to prepare a plurality of colloids; lectins are sequentially added into the colloids, different colloids correspond to different lectins in a one-to-one mode, and then streptavidin-peroxidase is added for incubation; and after incubation is finished, smearing the chemiluminescence liquid on the colloid, and determining the glycosylation type and quantifying the glycosylation abundance of the luminescence product through an imaging system. According to the present invention, the rapid detection of the VWF glycosylation mode is achieved, the sugar chain coverage range is expanded, the detection specificity is improved, and the observability of the sugar chain signal is improved.

The invention relates to the technical field of platelet lysate monitoring, and discloses a platelet lysate quality real-time monitoring device based on multi-modal sensing, which comprises a monitor main body, and a plurality of test tubes for bearing lysate samples are arranged in the monitor main body. A liquid storage cylinder for bearing pyrogen-free diluent and tachypleus amebocyte lysate is arranged above the test tube, an instillation mechanism for automatically instilling liquid in the liquid storage cylinder into the test tube is arranged in the monitor main body, and a shaking mechanism for shaking and mixing the test tube is arranged between the monitor main body and the liquid storage cylinder; according to the device, individual operation differences and visual fatigue errors which are difficult to avoid and exogenous pollution risks possibly introduced in the pipetting process, which are caused by manual one-by-one sample adding, are eliminated, and strict standardization and whole-course traceability of key parameters in the detection process are realized; therefore, the requirements of high throughput, high reliability and compliance of endotoxin detection in industrial production of the platelet lysate are met.

The invention belongs to the technical field of biological medicine, and particularly discloses injectable hydrogel based on platelet lysate modification, a preparation method of the injectable hydrogel and application of the injectable hydrogel in osteochondral defect repair. The platelet lysate modified injectable hydrogel is photosensitive dual-network hydrogel with a uniform structure, which is constructed by taking platelet lysate (PL), methacrylic anhydride (MA), E7 peptide and polyethylene glycol diacrylate (PEGDA) as raw materials through a photocuring technology, and the hydrogel is cured after in-situ injection at a defect position to promote osteochondral regeneration. The dual-network hydrogel can simultaneously meet the extracellular matrix bionic requirements of cartilage and bone due to the characteristics of rigidity and softness; the combination of the E7 peptide and the PL can effectively utilize a local microenvironment and promote migration and directional differentiation of surrounding stem cells. Therefore, the invention provides a self-adaptive hydrogel scheme which is simple and convenient in preparation process and can promote layered repair of osteochondral defects by using a surrounding matrix environment.

According to the hypothalamic stem cell culture medium and the preparation method thereof provided by the invention, the human platelet lysate is introduced into the basal culture medium for extracting and culturing the hypothalamic stem cells, and the human platelet lysate is long in activity maintenance time and has a remarkable effect on proliferation of the hypothalamic stem cells; the human-derived platelet lysate reduces the possibility of influence of exogenous protein, is rich in a large amount of growth factors, is simple in extraction process and small in loss, and can play a good role in primary extraction culture of cells as a culture medium additive.

Provided are a method for producing a platelet lysate, a production system, and a bag set capable of efficiently producing a platelet lysate with a simple device structure. According to the method for producing a platelet lysate, by which a platelet lysate (PL) that contains a growth factor contained in platelet is produced, buffy coat is collected from whole blood; the buffy coat (BC) is centrifuged to extract a supernatant, thereby collecting a platelet concentrate from which leukocyte has been removed; and, prior to freezing, the platelet concentrate is centrifuged to remove the supernatant, thereby preparing a highly concentrated platelet concentrate. The highly concentrated platelet concentrate is then frozen and thawed, and further centrifuged to recover the platelet lysate.

The application discloses a kind of cell cryopreservation solution and preparation method thereof, the cell cryopreservation solution includes glycerol, platelet lysate, proline, L-glutamine and support material, basal medium.Compared with prior art, the cell cryopreservation solution prepared by the application can provide the nutrients and growth factors required by cells, help maintain the stability and survival rate of cells, reduce the damage to cells during freezing and thawing, and improve the survival rate and viability of cells.

A dressing comprises a delivery vehicle and platelet lysate. The dressing can also include an antimicrobial agent, a hemostatic agent, and a binder. It is contemplated that the dressing can be used for expediting hemostasis, improving antimicrobial activity, minimizing fluid loss, and accelerating wound healing when applied to a wound. The dressing would be useful in applications including military in-theater medical care and for conditions such as diabetic foot ulcers, as well as other applications.