31 results about "Platelet lysate" patented technology

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 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.

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.

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.

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.

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.

The invention belongs to the technical field of biological product preparation, and particularly relates to a method for preparing a platelet lysate through combination of freeze thawing and ultrasound and application of the platelet lysate. The method provided by the invention comprises the following steps: S1, preparing and obtaining platelet-rich plasma (PRP) by taking umbilical cord blood as a raw material; s2, adding a protective agent into the platelet-rich plasma prepared in the step S1; and S3, preparing platelet lysate (HPL) by using a freeze thawing and ultrasonic combination method. The platelet membrane can be more easily cracked by freezing and thawing through a combined method and then carrying out ultrasonic treatment; according to the protective agent, glutathione and glycerol are added to inhibit active oxygen generated by destroying cell membranes due to the cavitation effect in the ultrasonic process and protein structure damage caused by ice crystal formation in the freezing and thawing process, so that the problems of incomplete platelet lysis, serious growth factor and protein loss and the like in the HPL preparation process are effectively solved, the operation is simple and convenient, and the cost is low. And the cost can be effectively saved.

The invention provides an isolated culture method, and belongs to the field of biology. The isolated culture method comprises the following steps: adding hydroxyethyl starch into the uterine blood sample and uniformly mixing to obtain a mixed solution; standing the mixed solution; when red blood cells are layered, sucking supernate; centrifuging the supernate to obtain a precipitate; resuspending the precipitate by using a complete medium containing platelet lysate to obtain a primary cell suspension; and culturing the primary cell suspension to obtain the uterine blood mesenchymal stem cells. According to the isolated culture method, hydroxyethyl starch is added into the uterine blood sample, erythrocytes in the uterine blood sample can be settled by the hydroxyethyl starch, the extraction rate of leukocytes in the uterine blood sample can be increased by combining with an optimized centrifugal process, the cells can be better centrifugally cleaned, and the separation time is short.

To provide a recovery bag set and a platelet lysate production method which are capable of efficiently producing a platelet lysate with a simple system structure.SOLUTION: A recovery bag set 16 includes: a PL separation bag 16a consisting of a connection tube 16e that receives the flow of highly concentrated platelet concentrate having undergone freezing and thawing processes, and a transfer port 16f that allows the outflow of platelet lysate separated via centrifugation; a transfer tube 16d connected to the transfer port 16f; and a PL recovery bag 16b that is connected to the PL separation bag 16a through the transfer tube 16d and recovers the platelet lysate.SELECTED DRAWING: Figure 1

The invention belongs to the technical field of cell culture, and discloses a bone marrow cell culture medium and a preparation method thereof, and a method for in-vitro culture of bone marrow cells, the bone marrow cell culture medium is composed of a basic culture medium, streptomycin, gentamicin, a platelet lysis buffer, and glutamate enzyme; the final concentration of streptomycin is 50-100 [mu] g / mL, the final concentration of gentamicin is 20-50 [mu] g / mL, the final concentration of platelet lysate is 5-15% (v / v), and the final concentration of glutamate enzyme is 0.1-1 mmol / L. The bone marrow cell culture medium can efficiently support in-vitro proliferation of bone marrow cells, through specific combination and appropriate concentration control of the culture medium, the medium-term cell acquisition amount can be remarkably increased, and medium-term cell chromosomes are complete in morphology and good in dispersity; the standard requirements of clinical bone marrow cell chromosome karyotype analysis, blood disease diagnosis and cell genetics experiments in the scientific research field can be met.

A method for generating platelet lysate from platelet-rich plasma (PRP) includes placing a predetermined volume of PRP into a centrifuge tube. The method further includes sealing the centrifuge tube and placing the sealed centrifuge tube containing the PRP into a water bath. The method further includes operating an ultrasonic generator to direct ultrasonic waves onto the PRP in the sealed centrifuge tube in the water bath for a predetermined amount of time, thereby generating platelet lysate from the PRP. The method further includes spinning the sealed centrifuge tube containing the platelet lysate in a centrifuge, thereby separating byproducts out of the platelet lysate.

The invention provides a composition and a method for culturing mesenchymal stem cells, the mesenchymal stem cells and application of the mesenchymal stem cells. Experiments prove that a culture medium comprising a basal culture medium, platelet lysate, heparin and a beta-FGF2 factor is suitable for culturing the mesenchymal stem cells, and a method for culturing the mesenchymal stem cells is developed on the basis of the culture medium. The mesenchymal stem cells obtained by the method disclosed by the invention are high in yield, purity and activity and have better immunosuppression capability. According to the method, the preparation cost and the damage to the cells are reduced, the high-quality mesenchymal stem cells can be obtained on a large scale through the method, and powerful support is provided for subsequent cell treatment, tissue repair and regenerative medicine application.

The application provides an inflammation response type biomimetic hydrogel as well as a preparation method and application thereof, and belongs to the biomedical field. The inflammation response type biomimetic hydrogel is prepared from crosslinked polymers, hyaluronic acid, platelet lysate and nanoparticles in an aqueous solution. The inflammation response type biomimetic hydrogel can be used as a scaffold to remodel a pathological environment, convert an infarction site into a state promoting regeneration, promote vascular regeneration, regulate the survival and differentiation of neural stem cells, and promote endogenous regeneration repair, and has a good application prospect in the treatment of ischemic stroke.

The application discloses a high-purity NKT cell preparation and a preparation method and a cryopreservation regulation method, and belongs to the fields of cell biology technology and immunotherapy. The method systematically solves multiple challenges of low purity, insufficient amplification efficiency, poor cell viability, high cost and difficulty in large-scale production of NKT cells in the prior art through a unique secondary cryopreservation process, innovative application of platelet lysate and specific induction of alpha-galactosylceramide (alpha-GalCer). The prepared NKT cell preparation has the advantages of high purity, large amplification fold, strong cell function activity and good safety, and provides a high-quality and standardized cell source for clinical applications such as tumor immunotherapy, and has great industrialization prospects.

The invention discloses a serum-free medium for resisting aging of mesenchymal stem cells, a preparation method of the serum-free medium and a mesenchymal stem cell culture method, and relates to the technical field of mesenchymal stem cell culture.The serum-free medium for resisting aging of mesenchymal stem cells comprises a basic medium and additives, the basic medium is a DMEM medium, the additive is prepared from ascorbic acid trisodium phosphate (AA2P), lecithin, biotin, tocopheryl acetate, sodium selenite, EUK134, glutathione, wheat protein hydrolysate, putrescine, AKT2Protein, wnt3a, oleic acid, linoleic acid, albumin and platelet lysate; the preparation method of the serum-free culture medium comprises the following steps: S1, preparing an albumin-fatty acid compound; s2, preparing a culture medium compound; by adopting the serum-free culture medium provided by the invention, cell senescence is effectively reduced, and the senescence process of MSCs can be effectively delayed; the proliferation speed of the MSCs can be obviously improved.

This invention discloses a method for preparing platelet lysis buffer and its application, belonging to the field of cell biotechnology. The invention employs X-ray irradiation inactivation technology, irradiating plasma supernatant at a dose of 25-35 kGy under low-temperature and light-protected conditions (2-8℃) to thoroughly inactivate immunologically active substances and pathogenic microorganisms, while avoiding thermal denaturation of growth factors. This is combined with segmented ultrasonic cyclic lysis and ice-water bath temperature control (≤37℃) to ensure complete platelet lysis and reduce damage to active factors. Further treatment with calcium gluconate removes fibrin, preventing the precipitation of flocculent material. The preparation method of this invention is process-controllable and highly safe, with a core growth factor retention rate ≥80% and a fibrin removal rate exceeding 92%. The prepared platelet lysis buffer can be added to mesenchymal stem cell culture medium, effectively meeting the high-quality requirements of clinical-grade cell culture.

The invention provides an optimized umbilical cord mesenchymal stem cell culture solution and a preparation method thereof, and relates to the technical field of cell culture, the optimized umbilical cord mesenchymal stem cell culture solution comprises a basic culture medium, a platelet lysate, a cell generation factor, L-alanyl-L-glutamine and a mixed solution; the mixed solution consists of an antioxidant solution, an adherent proliferation solution and a metabolic regulation solution. The selected basic culture medium can provide an adaptive basic nutrition environment for cells, the blood platelet lysate is matched to reduce exogenous pollution, and meanwhile, the blood platelet lysate and the cell generating factors have a synergistic effect, so that efficient proliferation of the cells is assisted, the dryness of the cells can be stably maintained, and the loss of a dryness marker in the passage process is avoided; by adding the L-alanyl-L-glutamine, the problem of ammonia toxicity of conventional glutamine can be avoided, stable cell metabolism is guaranteed, and the mixed solution can optimize a cell growth microenvironment from multiple aspects, assist cells to keep a good growth state, ensure high activity of the cells and reduce the risk of cell apoptosis.

The application relates to a preparation method and application of sPL gel for endometrial injury repair, relates to the field of endometrial repair drugs, and aims at solving the problems of short treatment time and poor treatment effect of existing platelet-rich plasma. Method: 1. preparing a platelet lysate PL; 2. diluting placental chorionic trophoblast cells with complete culture medium to obtain a cell suspension, culturing the cell suspension by adding bisabolene, collecting a culture solution, filtering the culture solution to obtain a conditioned medium, and mixing the platelet lysate PL and the conditioned medium to prepare sPL; 3. adding a collagen solution and carbomer to the sPL, uniformly mixing the sPL, collagen solution and carbomer to prepare sPL gel. The sPL gel prepared by the method can be solidified by temperature rise, effectively improves the internal microenvironment of the uterine cavity, and is used for promoting the repair of endometrial injury.