1885 results about "Stem cell" patented technology

A bioenergy generator to increase the life force of a biological system is provided. The bioenergy generator includes fine pieces of natural stone, fine pieces of natural sand, fine pieces of metal, water and a supporting material operable to hold the natural sand, the natural stone, the metal, and the water together. Uses of the bioenergy generator are also provided, in particular, to enhance stem cell production.

The invention discloses a culture medium and application thereof, and a method for in-vitro separation culture and adipogenic differentiation of precursor adipocytes in goat muscle, and relates to the technical field of cell culture, the culture medium comprises the following components: 8-12 [mu] g / mL of insulin, 8-12% by volume of fetal calf serum, 95-105 U / mL of penicillin, 95-105 [mu] g / mL of streptomycin and the like. On the basis of the prior art, a proper amount of liquiritin and pilocarpine are mainly added into a culture medium, and synergistic cooperation of the liquiritin and pilocarpine is found, so that the adipogenic differentiation capacity of cells such as mesenchymal stem cells and precursor adipocytes is remarkably improved, and the application range is wide. On the basis, the invention also provides a complete set of perfect in-vitro isolated culture and adipogenic differentiation method of the goat intramuscular precursor adipocytes, and a method and a way are provided for a series of researches developed through the goat primary intramuscular precursor adipocytes in the future.

The invention relates to the technical field of stem cells, in particular to a lentivirus envelope plasmid combination and application thereof, a lentivirus and a packaging method thereof and a method for transduction of hematopoietic stem cells. The invention provides an envelope plasmid combination for lentivirus packaging, which is composed of lentivirus packaging plasmids containing VSVG glycoprotein and lentivirus packaging plasmids containing BaEV glycoprotein in a ratio of 3: 7. The invention further provides a method for transduction of the hematopoietic stem cells by the lentivirus, the method is simple and convenient to operate, the long-term dryness of the hematopoietic stem cells in vitro can be maintained, efficient and stable transduction of the hematopoietic stem cells can be realized, and the transduction rate is greater than 90%. The lentivirus transduction method hematopoietic stem cells can be used for hematopoietic stem cell gene therapy hematopoietic system genetic diseases, such as severe combined immunodeficiency, beta-thalassemia and sickle cell disease, Wiskott-Aldrich syndrome and the like, and the application prospect is good.

The present disclosure relates to cellular compositions that are modified to introduce a nucleic acid or vector expressing the same. Such compositions may be used to deliver nucleic acid to a target cell and treat disease such as cancer.

The invention belongs to the field of antibody development, and discloses a screening method and application of a human-mouse protein high homologous target antibody based on a fully humanized antibody mouse. Aiming at the problem of weak antibody response caused by immune tolerance of human-mouse high homologous targets (protein homology is greater than or equal to 95%), the following scheme is provided: in embryonic stem cells (ES cells) of HUGO-Mabfully humanized antibody transgenic mice, a mouse target gene (such as ACVR2A) is knocked out through a Turbo Knockout technology, and homozygous knockout ES clones are screened; carrying out microinjection on the clones to the whitened B6 mouse blastocyst, and transplanting a pregnant mouse to obtain a Founder mouse; the Founder mouse is subjected to target antigen immunization for more than or equal to 4 times (the Freund's complete adjuvant is used for the first time), and the titer of the serum antibody is detected. According to the invention, 100% homozygous knockout chimeric efficiency is realized in the Founder stage, the mouse construction period is shortened from traditional 8-10 months to 3-4 months, and the diversity and affinity of the antibody are significantly improved (titer reaches 1: 729,000). The obtained antibody can be used for preparing medicines for treating tumors or autoimmune diseases.

The invention provides a pancreatic islet organ capable of efficiently maintaining pancreatic islet beta cell identity and a preparation method and application thereof. Specifically, the invention provides the preparation method for the in-vitro pancreatic islet organ, and the preparation method comprises the following steps: carrying out induced differentiation culture on the ZnT8-knocked-out pluripotent stem cells, so as to obtain the mature pancreatic islet organ. The invention also provides a corresponding transplantation composition and application thereof. The invention discloses that ZnT8 knockout is a key factor for effectively maintaining the identity stability of beta cells in pancreatic islet organs. In addition, the pancreas islet beta cell identity maintenance agent is combined with the ZnT8-knocked-out pancreas islet organ for use, so that the identity stability of the pancreas islet organ beta cell can be obviously maintained. The invention provides a new strategy for transplantation treatment of type 2 diabetes, and has excellent clinical transformation value.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

The invention relates to injectable temperature-sensitive hydrogel loaded with traditional Chinese medicine active exosomes, and belongs to the technical field of clinical treatment of diabetic foot refractory wounds, and the injectable temperature-sensitive hydrogel is obtained by poloxamer 407 hydrogel and a hyaluronic acid loaded composite carrier; the composite carrier body is formed by coating a traditional Chinese medicine active exosome with a coating solution; the traditional Chinese medicine active exosome is obtained by inducing adipose-derived mesenchymal stem cells with a modified astragalus extract. The injectable temperature-sensitive hydrogel is obtained by loading a composite carrier with poloxamer 407 hydrogel and hyaluronic acid, the poloxamer 407 and the hyaluronic acid quickly form a physical barrier on a wound surface after being injected, a moist healing environment is provided, and continuous and slow release of exosome and active ingredients is realized by means of the temperature-sensitive characteristic and the network structure of the poloxamer 407 and the hyaluronic acid, so that the wound healing effect is improved. And over-high local concentration or premature loss caused by sudden release of the medicine is avoided.

The invention discloses a function-enhanced mesenchymal stem cell preparation obtained by utilizing arthritis effusion and hypoxia pretreatment of mesenchymal stem cells as well as a preparation method and application of the function-enhanced mesenchymal stem cell preparation, and relates to the technical field of biological medicines. The preparation method of the function-enhanced mesenchymal stem cell preparation obtained by utilizing arthritis effusion and hypoxia pretreatment of mesenchymal stem cells comprises the following steps: S1, obtaining mesenchymal stem cells and performing hypoxia amplification passage; s2, pretreatment of arthritis effusion; S3, pre-stimulation of the arthritis effusion of the mesenchymal stem cells to obtain function-enhanced mesenchymal stem cells; s4, index detection and screening; s5, preparing a function-enhanced mesenchymal stem cell preparation; optimized low-oxygen pretreatment is adopted, so that the cell activity is improved; the mesenchymal stem cells subjected to joint pretreatment of arthritis effusion and hypoxia enhance expression and secretion of anti-inflammatory factors and immunomodulatory molecules in the cells; the pre-stimulated mesenchymal stem cells show a stronger immunosuppression function in vitro.

The invention discloses a method for quickly and efficiently differentiating iPSC (induced pluripotent stem cells) into mesenchymal stem cells in a serum-free manner, the method comprises the following steps: firstly inducing the iPSC into a mesoderm precursor, then transferring the mesoderm precursor into a liquid culture medium for suspension culture, and finally obtaining iMSC (iPSC-doped MSC) by using an enrichment culture medium, and the method can shorten the differentiation time, improve the purity of the iMSC and obtain a large number of homogenized iMSCs.

The invention discloses an adipose-derived stem cell exosome compound loaded with silver nanoparticles and application of the adipose-derived stem cell exosome compound loaded with the silver nanoparticles, and ADSC-Exos loaded with the silver nanoparticles is developed by integrating AgNPs into an exosome of ADSCs. In vivo, the Exo-AgNPs can accelerate healing of infected wounds by reducing bacterial colonization and promoting collagen deposition, angiogenesis and M2 type macrophage polarization. In vitro, the Exo-AgNPs activates the functions of fibroblasts and vascular endothelial cells, and shows an excellent antibacterial property. In addition, the Exo-AgNPs induces polarization of M2 type macrophages and inhibits secretion of proinflammatory cytokines by activating autophagy. The results show that Exo-AgNPs is expected to become an ideal biological material for treating infected wounds.

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and / or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

The invention discloses a temperature-sensitive hydrogel loaded with mesenchymal stem cell exosome as well as a preparation method and application of the temperature-sensitive hydrogel, and belongs to the technical field of biological medicines. The method comprises the following steps: (1) adding poloxamer 407 into a pre-cooled dispersion medium to enable the final mass concentration to be 20-30%, and oscillating and dissolving at 4 DEG C overnight to obtain poloxamer hydrogel; (2) collecting a mesenchymal stem cell culture supernatant, and then centrifuging, wherein the centrifuging condition is 200 * g * 15 minutes to 2000 * g * 20 minutes to 12000 * g * 20 minutes; filtering the supernatant, concentrating by using an ultrafiltration tube, further performing ultracentrifugation on the concentrated solution, re-suspending the precipitate by using DPBS, centrifuging again, and re-suspending the finally obtained exosome precipitate by using DPBS to obtain an exosome suspension; the ultracentrifugation condition is as follows; and (3) taking the poloxamer hydrogel prepared in the step (1), adding the exosome suspension obtained in the step (2) under an ice-water bath condition, and uniformly mixing to obtain the poloxamer temperature-sensitive hydrogel.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

The invention belongs to the technical field of exosome freeze-dried powder, and discloses exosome freeze-dried powder and a preparation method thereof, the exosome freeze-dried powder contains mesenchymal stem cell exosome, trehalose, sodium hyaluronate, hydroxypropyl-beta-cyclodextrin, glutathione, ceramide and fructo-oligosaccharide. According to the exosome freeze-dried powder, hydroxypropyl-beta-cyclodextrin, glutathione, ceramide and fructo-oligosaccharide are added, the membrane structure integrity of the exosome is effectively protected, the retention rate of active substances in the exosome freeze-dried powder is high, and therefore the effects of resisting inflammation, regulating cell metabolism, promoting tissue repair and the like of the exosome can be fully exerted. In addition, glutathione can directly remove skin free radicals and inhibit oxidative stress injury, and has an anti-oxidation effect; ceramide can also supplement skin lipid barrier, enhance the water locking capacity and enhance the moisturizing effect of sodium hyaluronate; fructo-oligosaccharide can also regulate skin micro-ecological balance, and has anti-inflammatory and bacteriostatic effects.

The invention relates to a composite hydrogel artificial bile duct as well as a preparation method and application thereof, the artificial bile duct is prepared by taking sodium alginate (SA) loaded with human umbilical cord mesenchymal stem cells and methylacryloylated gelatin (GelMA) composite hydrogel as raw materials, and the artificial bile duct can be used for biological materials or instruments for treating bile duct defects. The composite hydrogel artificial bile duct has better biocompatibility, can be tightly connected with an in-vivo tissue mucous membrane, and reduces stimulation to a local mucous membrane, so that bile duct obstruction caused by local tissue granulation hyperplasia is avoided, and adverse reactions are reduced; in addition, the composite hydrogel artificial bile duct can be automatically degraded after being implanted into a human body; the structure is compact, the mechanical property is good, and a good environment is provided for cell proliferation; the human umbilical cord mesenchymal stem cells can be directionally differentiated into bile duct cell-like cells, and bile duct injury repair is promoted.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some embodiments, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some embodiments, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

The invention provides a method for overspeed preparation of gene edited goldfish. The method comprises the following steps: S1, obtaining gobiocypris rarus without primordial germ cells as receptor fish; s2, designing a target spot for the goldfish amh gene, introducing a Cas9 / gRNA complex into a goldfish fertilized egg, performing sequencing verification to obtain a mutant individual, and feeding the mutant individual as a donor fish; s3, taking gonads of donor fish, cutting into pieces, performing enzymolysis digestion, filtering, and performing density gradient centrifugation to enrich donor germline stem cells; s4, transplanting the germline stem cells of the donor fish into the receptor fish body, and feeding to obtain the gene edited goldfish sperms and ova. S5, hybridizing and feeding goldfish amh mutant sperms and ova produced by gobiocypris rarus to obtain gene edited goldfish individuals. According to the invention, gobiocypris gobiocypris is used as a receptor, the cross-subfamily gene edited goldfish is used as a donor, and an efficient, stable and easy-to-popularize method for preparing the gene edited goldfish is provided by utilizing a belly borrowing reproduction technology.

The invention discloses HA-PAS hydrogel for enhancing cartilage repair of early osteoarthritis and a preparation method of the HA-PAS hydrogel. According to the hydrogel, formylated hyaluronic acid, hydrazide modified low-methoxyl pectin and phenylboronic acid modified alginate form a ternary polysaccharide skeleton, a double-dynamic cross-linked network is formed through acylhydrazone bonds and boric acid ester bonds, and a triple network structure is formed by combining permanent cross-linking catalyzed by horse radish peroxidase. The hydrogel integrates functional components such as nano-selenium, a pulullan-strontium chelate and a mesenchymal stem cell source exosome, so that multiple effects of resisting oxidation, resisting inflammation, promoting cartilage differentiation and repairing subchondral bones are achieved. The compression modulus of the hydrogel is 0.8-1.5 MPa, and the hydrogel can be injected for in-situ gelling and MMP responsive degradation, shows an excellent cartilage repair effect in animal experiments, and provides a new solution for treatment of early osteoarthritis.

The invention discloses a special-shaped titanium alloy capable of enhancing bone regeneration and repair efficacy as well as a preparation method and application of the special-shaped titanium alloy. The special-shaped titanium alloy comprises a special-shaped titanium alloy base body and a porous structure attached to the special-shaped titanium alloy base body, and the porous structure is in multi-stage size distribution and comprises long-strip-shaped gullies (100-150 micrometers wide) and large holes (30-50 micrometers wide). And the morphology in the gullies shows coralline ligaments and a nest-shaped pore channel (0.5-2 microns) structure. This can provide space and sites for the growth and adsorption of osteoblasts. According to the method, a gas phase alloying (VPA) and VPD synergistic process is adopted in a breakthrough mode, a non-oxidation / nitridation coralline-shaped hierarchical pore structure is constructed on the surface of the commercial titanium implant, and it can be achieved through verification of an aging animal model that a special-shaped matrix is uniformly modified in the whole domain, the regeneration function of aging stem cells is activated, and the bone repair and regeneration effect is effectively improved.

The invention provides a nerve growth factor functionalized exosome as well as a preparation method and application thereof. The functionalized exosome can activate a Wnt / beta-catenin signal channel to maintain dryness of corneal limbal stem cells (LSCs) so as to promote corneal epithelium repair, can activate a cAMP signal channel to relieve inflammatory response and promote corneal nerve regeneration, and can be used for treating or relieving corneal chemical burns, including relieving inflammatory response, reducing stromal scar formation and inhibiting corneal neovascularization. Corneal epithelium repair and nerve regeneration are promoted.

The invention relates to the technical field of biology, in particular to application of active polypeptide in maintaining dryness of umbilical cord mesenchymal stem cells. The active polypeptide disclosed by the invention is PNC-27, and the amino acid sequence of the PNC-27 is as shown in SEQ ID NO: 1. Experiments find that the PCN-27 polypeptide can effectively promote proliferation of the umbilical cord mesenchymal stem cells by regulating expression of periodic proteins Cyclin-D1 and CDK1, and can also effectively maintain dryness of the umbilical cord mesenchymal stem cells by promoting expression of dry gene Oct-4 protein in the umbilical cord mesenchymal stem cells. Experiments fully prove that the PCN-27 polypeptide can effectively maintain the dryness of the umbilical cord mesenchymal stem cells and stimulate the potential application value of the umbilical cord mesenchymal stem cells in the fields of cell culture, biomedicine and the like.

The invention relates to a serum-free culture medium for culturing mesenchymal stem cells as well as a preparation method and application of the serum-free culture medium. Wherein the serum-free culture medium comprises a complete culture medium and praeruptorin E, and the content of the praeruptorin E in the serum-free culture medium is 0.25-3 [mu] M. The inventor finds that a serum-free culture medium containing a certain concentration of praeruptorin E can effectively improve the decline of replicative activity of mesenchymal stem cells in in-vitro passage. Specifically, the serum-free culture medium containing the praeruptorin E with a certain concentration can reduce the expression level of a marker beta-galactosidase with reduced replicative activity in the mesenchymal stem cells with reduced replicative activity after multiple in-vitro passage, reduce the expression of a gene CDKN1A with reduced replicative activity, and increase the multiplication capacity of the mesenchymal stem cells; the mitochondrial damage is reduced.

The invention belongs to the technical field of biomedicine, and relates to a method for increasing the yield of pancreas islet organs differentiated by pluripotent stem cells and improving the stability. The method comprises the following steps: (1) inoculating pluripotent stem cells, and culturing by adopting a basic culture medium containing Y-27632; (2) changing the liquid, and continuously culturing by adopting a basic culture medium; (3) culturing for 1 day by using an S1D1 culture medium, and then culturing for 2-3 days by using an S1D2 culture medium; (4) changing the liquid, and culturing for 2-3 days by adopting the S2 culture medium; (5) changing the liquid, and culturing for 1-2 days by adopting an S3 culture medium; (6) changing the liquid, and culturing for 4-7 days by adopting a culture medium S4; (7) changing the liquid, and culturing for 3-10 days by adopting an S5 culture medium; and (8) carrying out cell sphere repolymerization, and culturing for 3-21 days by using an S6 culture medium to obtain the pancreas islet organ. The present invention increases the yield and differentiation efficiency at each differentiation stage and increases the scale yield at the cellular level.

The application relates to a cartilage organoid based on a DNA-silk fibroin hybrid hydrogel sustained-release system and a preparation method and application thereof. The DNA-silk fibroin hybrid hydrogel sustained-release system is a hydrogel system after a DNA-silk fibroin hybrid hydrogel sustained-release system premixing solution is printed through a digital light processing system, wherein the DNA-silk fibroin hybrid hydrogel sustained-release system premixing solution contains DNA, silk fibroin, acrylated RGD peptide, acrylated polyethylene glycol NHS ester, glucosamine, TD-198946 and 2,4,6-trimethylbenzoyl lithium phosphate. Compared with the prior art, the glucosamine and TD-198946 introduced in the DNA-silk fibroin hybrid hydrogel sustained-release system can significantly improve the cartilage differentiation efficiency of bone marrow mesenchymal stem cells; and the application can be transplanted to a cartilage defect as a cartilage graft to promote cartilage regeneration and repair.

Radiotherapy is the most widely used and effective anti-tumor therapy, however it can damage healthy tissues surrounding the tumor. One approach to reducing damage to healthy tissue involves irradiation at dose rates far exceeding those currently used in clinical contexts reduces radiation-induced toxicities while maintaining an equivalent tumor response. This is known as the FLASH effect. The mechanism responsible for reduced tissue toxicity following FLASH radiotherapy (FLASH-RT) is still undetermined; multiple hypotheses have been suggested by linking the high dose rate to rapid oxygen depletion, immune response, reduction of peroxyl radical lifetime, preservation of normal tissue stem cells, etc. While effective, proton therapy is underutilized, contributing to fewer than 2% of all external beam radiation treatments. To overcome this shortcoming, provided are methods which utilize the highest energy layer and a universal range shifter (URS) to facilitate rapid spot arc therapy (RAPIDSPARC™).

The invention discloses application of a 3D hypoxia mesenchymal stem cell derived exosome preparation in preparation of an immune cell proliferation inhibitor or a medicine for treating acute myocardial infarction, belongs to the technical field of biology, and solves the problems of low cell survival rate and immunological rejection in application of a cell treatment method in the prior art. The 3D hypoxia mesenchymal stem cell derived exosome is used as a medium for treating acute myocardial infarction, and the problem of immunological rejection can be solved.

The application provides an improved hyaluronic acid injection preparation for inflammation targeting and stem cell capture, a preparation method and use thereof, and belongs to the field of medicine. The injection preparation is prepared from unsaturated bond modified hyaluronic acid, stem cell targeting DNA tetrahedron and thiol modified inflammation targeting aptamer as raw materials. The injection preparation can be targeted to accumulate at an osteoarthritis site, capture mesenchymal stem cells in situ, recruit mesenchymal stem cells to the osteoarthritis site, promote differentiation of stem cells into cartilage, promote cartilage repair, play a role in treating osteoarthritis, and has a good application prospect.

The application discloses a flavonoid compound in eaglewood as well as a preparation method and application of the flavonoid compound, and relates to the technical field of medicines. The application extracts and separates a new compound from natural eaglewood, and the new compound is used in the preparation of anti-osteoporosis drugs. The compound is detected in vitro to obtain an induction effect of the compound on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts, so as to affect the formation of osteoclasts. Meanwhile, the compound can promote the bone development of zebra fish through RT-qPCR detection, which indicates that the compound has anti-osteoporosis activity. The separation method of the compound is simple and easy to operate.

The invention relates to a bioactivity-related characteristic marker of umbilical cord mesenchymal stem cell-derived small extracellular vesicles and application of the bioactivity-related characteristic marker. The bioactivity-related characteristic marker of the umbilical cord mesenchymal stem cell-derived small extracellular vesicles comprises one or a combination of more than two of a PSMB3 protein, an SAA4 protein, a VPS33B protein and an AKR7A2 protein. The invention provides a screening method of the characteristic markers, and particularly, the biological activity of the human umbilical cord mesenchymal stem cell source extracellular vesicles can be effectively judged by detecting the characteristic markers or the combination of the characteristic markers of the umbilical cord mesenchymal stem cell source extracellular vesicles. An evaluation means and an important reference are provided for product quality control, clinical transformation and the like based on the umbilical cord mesenchymal stem cell-derived small extracellular vesicles.