66 results about "Platelet lysate" patented technology

The invention relates to the technical field of NK cell culture, and discloses a serum-free medium for NK cells from umbilical cord blood after cryopreservation and a culture method thereof.The medium comprises a coating solution, an induction medium and an amplification medium; wherein the coating liquid is prepared from an anti-CD16 monoclonal antibody and DPBS (diphenylphosphonic acid); the induction culture medium comprises a basic culture medium, a human-derived platelet lysate and cell factors IL-15, IL-18 and IL-21; the amplification culture medium comprises a basic culture medium, a human-derived platelet lysate and IL-2. The culture method comprises the following steps: obtaining mononuclear cells from fresh umbilical cord blood, and then cryopreserving and resuscitating; and carrying out multiplication culture on the recovered umbilical cord blood mononuclear cells by using the serum-free culture medium. According to the NK cell culture medium, the NK cells with high purity, high killing activity and high cytotoxic activity can be obtained without autoserum culture.

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.

Disclosed are methods for treating platelet compositions (e.g. platelet concentrates and / or platelet lysates) with electron beam radiation, where the compositions are in a frozen state during irradiation with the e-beam radiation. The methods can be conducted using e-beam radiation at doses effective to reduce the pathogen content of the compositions while retaining highly beneficial bioactivities of the compositions. Also disclosed are compositions preparable by the methods, and methods and compositions involving the use of the e-beam treated materials.

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 generally relates to the field of cell culture technology, and specifically, to a culture medium and a culture method for infrapatellar fat pad stem cells. The infrapatellar fat pad stem cell culture medium comprises a basal culture medium, and working concentrations of 3v / v%-7v / v% platelet lysate, 10ng / mL-30ng / mL TGF-β1, 0.5μmol / L-1.5μmol / L dexamethasone, and 0.1μg / mL-1μg / mL non-cytokine proinflammatory factors. The culture medium can promote the growth of IPFP-ASCs and has a significant promoting effect on the in vitro cartilage differentiation ability of IPFP-ASCs.

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 provides a method for efficiently separating and culturing mesenchymal stem cells. The method comprises the steps of separating adipose-derived mesenchymal stem cells, culturing the cells and the like. In the steps of adipose tissue treatment and cell separation, a specific digestive enzyme mixed solution is used for digestion treatment, so that the adipose tissue is fully digested, the cell damage is reduced to the greatest extent, the separation of adipose cells is promoted to the greatest extent, and the yield of the cells is remarkably improved. For cell culture, a serum-free culture medium is innovatively adopted. The formula of the culture medium is elaborately blended, a DMEM-F12 culture medium is used as a matrix, and platelet lysate, human serum albumin, recombinant insulin, EGF, bFGF, special oligopeptide and other components in a specific proportion are added. The components have a synergistic effect, so that the potential risk caused by serum is avoided, a suitable growth environment is provided for cells, the growth and proliferation of the cells are promoted, and the undifferentiated state of the cells is maintained.

The invention discloses a TIL cell culture medium and a TIL cell culture method, an activation and proliferation culture medium of TIL cells comprises the following components: a T cell serum-free culture medium, IL-2, platelet lysate and gentamicin, a CD3 monoclonal antibody, a CD28 monoclonal antibody and phytolectin PHA-P are added for cell activation, and then the activation and proliferation culture medium is used for activation and proliferation culture. Wherein CD3 and CD28 antibodies are used in combination to stimulate TIL cells and simulate the double-signal effect of in-vivo TIL cell activation, meanwhile, PHA-P is utilized, by means of the characteristics of PHA-P, small gonorrhea cells are stimulated in vitro to be converted into gonorrhea cells with immunocompetence, division of the gonorrhea cells is promoted, and the proliferation capacity of the TIL cells is improved through the synergistic effect of PHA-P and IL-2; platelet lysate is used for replacing plasma of a patient and is used as an efficient culture supplement for supporting TIL cell growth; finally, more TIL cells are obtained through the technology, and the number of immune cells required by clinical effective treatment can be sufficiently met.

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.

The present invention relates to compositions, kits and methods for providing mechanical support and / or medicaments to biological targets. Specifically, the compositions include platelet lysates, fibrinogen and thrombin. The compositions can be used to treat wounds / injuries (e.g., cornea) and for other applications involving tissue cultures.

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 provides a keratin 15 type hair follicle stem cell culture medium, an adherent culture method and application, and belongs to the technical field of stem cell culture. The invention provides a keratin 15 type hair follicle stem cell culture medium, which comprises a basic culture medium, a serum substitute and a nutrition additive, a mixed culture medium containing a DMEM basic culture medium and an F12 basic culture medium is used as the basic culture medium, and an epidermal growth factor, a platelet lysate and a bovine pituitary extract are used as the serum substitute. The keratin 15 type hair follicle stem cells are cultured on the basis of the culture medium, proliferation-terminated autologous mesenchymal cells serve as a feeding layer, the activity of the hair follicle stem cells p63 can be reserved, and the keratin 15 positive hair follicle stem cells are amplified.