239results about "Cell membrane modification" patented technology

The invention relates to the technical field of biological medicines, in particular to an iRGD targeted and anti-fibrosis small molecule dual-modified exosome as well as a construction method and application thereof. The exosome utilizes the specific binding penetration enhancement effect of iRGD (the amino acid sequence is CRGDKGPDC) and a fibroblast high-expression integrin receptor, so that the enrichment efficiency of the exosome at a focus part is improved; the anti-fibrosis micromolecule Let-7a-5p can inhibit a TGF-beta signal channel by blocking Smad2 / 3 phosphorylation, and the anti-fibrosis micromolecule Let-7a-5p and the anti-fibrosis microRNA carried by the exosome generate a synergistic anti-fibrosis effect; the exosome phospholipid bilayer can also protect Let-7a-5p from enzymolysis and prolong the half-life period, and iRGD modification can reduce the non-targeted uptake rate. An animal model verifies that the double-modified exosome has a good skin fibrosis resisting effect, and side effects such as liver and kidney function damage are not found. The invention provides an effective and safe treatment scheme for resisting skin fibrosis.

The invention provides brain cell specific targeting delivery based on exosome, and the exosome secreted by 293T cells is used for engineering transformation, so that the capabilities of penetrating BBB and specifically targeting neurons are achieved. According to the present invention, the L1CAM is loaded on the exosome, the exosome is obtained by transfecting a 293T cell of a Lamp2b-L1CAM PCDH-HygroHA plasmid through a 293T-Lamp2b-L1CAM cell line, and the BBB cell-penetrating peptide 4F-T7 and the neuronal targeting protein L1CAM are loaded on the surface of the exosome so as to treat the neurodegenerative diseases such as the Alzheimer's disease and the Parkinson's disease.

The invention discloses a prebiotic-loaded intestinal targeting exosome and a preparation method and application thereof.The preparation method comprises the steps that firstly, plasmids containing intestinal cell targeting peptide sequences are constructed through a gene editing technology, the plasmids are transfected and introduced into cells, the transfected cells are screened with antibiotics, and cell strains stably expressing intestinal cell targeting peptides are obtained; then culturing and adding prebiotics, and continuously culturing to enable the cells to take in the prebiotics and release the exosomes loaded with the prebiotics; and finally, collecting cell culture supernate, and centrifuging and resuspending for multiple times under a low-temperature condition to obtain the prebiotics-loaded intestinal targeting exosome. According to the preparation method, extraction of the exosome, loading of the prebiotics and an intestinal targeting modification process are optimized, so that the prepared intestinal targeting exosome loaded with the prebiotics can be used for remarkably improving the loading efficiency and intestinal targeting of the prebiotics and enhancing the stability of the exosome in gastrointestinal tracts; therefore, the delivery efficiency and the treatment effect of the prebiotics in the intestinal tract are improved.

The present invention provides compositions and methods for inducing antigen-specific tolerance in a subject. In one embodiment, the present invention provides a composition comprising an apoptotic body and an epitope of an antigen. Also provided herein are methods of preparing and administering the composition. The composition and methods provided herein can induce antigen-specific tolerance in a subject.

The invention relates to a polypeptide derivative for pancreas islet surface modification and inhibition of pancreas islet transplantation IBMIR reaction, and belongs to the technical field of biological medicine. The peptide derivative disclosed by the invention is obtained by carrying out an amide reaction on KF7 peptide and levodopa, and the amino acid sequence of the KF7 peptide is shown as SEQ ID NO: 1. The polypeptide derivative has good biocompatibility, the surface modification method is simple, the modification process has no influence on cell life activity, activation, aggregation, expansion and adhesion of platelets can be inhibited, the bleeding risk is reduced, a blood coagulation system is not influenced, pancreas islet intrahepatic colonization and pancreas islet functions after portal transplantation are enhanced, and the application prospect is wide. The composition can protect pancreas islet, prevent infiltration of platelets and CD11b + cells, inhibit pancreas islet transplantation IBMIR reaction, control blood sugar level, and prevent and treat diabetes mellitus.

The invention relates to a compound shown in a general formula I or a stereoisomer, a solvate, a hydrate, a prodrug, a stable isotope derivative and a pharmaceutically acceptable salt thereof. The compound can be anchored to the surface of a living cell through a hydrophobic acting force between a hydrophobic tail chain and a cell membrane and a receptor-ligand acting force between a ligand branch chain and cell membrane protein, the anchoring efficiency is high, the anchoring stability is high, and the treatment effect of the living cell on diseases can be enhanced; meanwhile, a biological orthogonal group can be introduced into the surface of a cell, and the cell anchored by the compound can be coupled with a nano-drug, a protein drug or a fluorescent probe and the like through a biological orthogonal reaction, so that the treatment effect on diseases can be remarkably improved, and in-vivo and in-vitro tracing of living cells can be realized. The compound shown in the general formula I provides a new technical platform for cell-based immune combination therapy, and provides a new thought and a new medicine for cell therapy based on various diseases. # imgabs0 #

The present disclosure relates to a mammalian cell which is modified to express on the surface of its membrane a binding domain which binds to a target molecule. The disclosure also relates to protein constructs and nucleic acids for producing such modified mammalian cells, and to methods for using the mammalian cells to deliver therapeutic agents to target cells or tissues in vivo.

The invention discloses a stem cell composition and application thereof in regulating sub-health. The stem cell composition comprises mesenchymal stem cells (MSCs) modified by fusion polypeptide and an anti-IL-6R single domain antibody. The amino acid sequence of the fusion polypeptide is as shown in SEQ ID NO.1, and the fusion polypeptide is modified on the surface of stem cells through click chemistry, so that the chemotactic migration ability, collagen matrix affinity and inflammation tolerance of the stem cells are remarkably improved. The anti-IL-6R single-domain antibody is obtained through phage display screening, the VHH gene of the anti-IL-6R single-domain antibody is shown as SEQ ID NO.2, and the anti-IL-6R single-domain antibody has high affinity and activity of inhibiting an IL-6 / STAT3 signal channel. Animal experiments show that the composition can significantly reduce inflammatory factor (IL-6) and cortisol levels of chronic stress model mice, improve oxidation resistance (SOD) and recover immune homeostasis, and provides an innovative treatment strategy for adjustment of sub-health status.

The present invention provides a method for safely and simply performing treatments such as exfoliation / floating treatment, dormancy treatment (dormancy not only enhances the protective effect but also suppresses unnecessary differentiation of cells, thereby maintaining an undifferentiated / low-differentiated state), and protective treatment for storage and transportation, on cells in tissue or cultured cells, while preventing their death and maintaining their viability, and a method for safely and simply performing cell activation treatment for implanting dormant floating cells. The cell treatment agent contains, as an active ingredient, at least one selected from alginic acid, heparins, sulfated dextran, and protease inhibitors.

Provided are: a culture medium which is for producing an organoid and which comprises a prostaglandin, hepatocyte growth factor (HGF), an interleukin (IL)-6 family protein, and an epidermal growth factor (EGF) family protein; a kit for cultivating an organoid; a method for producing an organoid; and an organoid.

The invention discloses an auxiliary protein composition, iron oxide nanoparticles, a preparation method and application, the composition comprises an SC protein with an amino acid sequence as shown in SEQ ID NO: 1 and an ST protein with an amino acid sequence as shown in SEQ ID NO: 2, and the composition can be used for modifying the iron oxide nanoparticles. The auxiliary protein composition can effectively promote endocytosis of the iron oxide nanoparticles, and assists the iron oxide nanoparticles to realize lysosome escape; moreover, the iron oxide nanoparticles modified by the auxiliary protein composition are good in biocompatibility and long in retention and storage time in cells, can be used for long-time labeling of the cells, and have wide clinical transformation and application prospects as a cell tracer agent.

The invention discloses an engineered exosome targeting liver blood sinus endothelial cells as well as a preparation method and application thereof, belongs to the technical field of biological medicines, and aims to solve the problem of lack of targeted LSEC drug delivery system research at present. The preparation method comprises the following steps: step 1, constructing a CD47OE-bEnd.3 stably transfected cell strain by utilizing lentivirus transfection of over-expressed CD47, and separating the exosome ExoCD47 from the CD47OE-bEnd.3 stably transfected cell strain; step 2, constructing a CD47OEWnt2OE-bEnd.3 stably transfected cell strain through Wnt2 lentivirus transfection, and separating an exosome Wnt2 (at) ExoCD47 from the CD47OEWnt2OE-bEnd.3 stably transfected cell strain; and step 3, modifying the membrane surface of the exosome Wnt2 (at) ExoCD47 obtained in the step 2 by using DMPE-PEG2K-CRLTRKRGLK, so as to obtain the RLTR-Wnt2 (at) ExoCD47. The RLTR-Wnt2 (at) ExoCD47 is an'eat me / dont eat me 'dual-targeting system, can effectively promote exosome targeting to LSEC, meanwhile, significantly inhibits phagocytosis of MPS, targets the LSEC to deliver Wnt2 mRNA, promotes the LSEC to release Wnt2 to a blood sinus microenvironment, activates a Wnt / beta-catenin signal channel, and further promotes proliferation of liver cells.

The present invention relates to the use of PEG-phospholipid molecules to selectively mask surface antigens on erythrocytes, thrombocytes and / or endothelial cells. Blood products treated with PEG-phospholipid molecules can thereby be infused into uncrossmatched or incompatible recipients with reduced risk of antibody binding to the surface antigens and agglutination. Correspondingly, organ transplants treated with PEG-phospholipid molecules can be transplanted into uncrossmatched or incompatible recipients with reduced risk of antibody binding and antibody-mediated rejection.

The application provides a preparation method of organoids based on a single cell chip, which comprises microfluidic chip preparation, a two-water-phase solution preparation, a single cell suspension preparation, microfluidic manipulation, single cell-loaded microgel generation and organoid construction and the like. The application takes a microfluidic chip integrated with a normally closed pneumatic pump valve as a technical platform, takes a single cell suspension pre-incubated with a crosslinking agent as a chemical reaction core, takes a water-in-water droplet as a forming template, and realizes efficient in-situ loading of single dispersed seed cells in a micro hydrogel carrier through a one-step method, and simply removes empty droplets without cells in a subsequent transfer step. The single cells in the microgel can be cultured, expanded and self-organized to form required organoids. The application improves the consistency of the starting point of the organoids, has the high-throughput characteristics of the droplet microfluidic technology, realizes efficient and highly reproducible in-vitro construction of the organoids, and can play a great role in the fields of basic research and transformation application related to the organoids.

[Problem] To provide a method for producing a dendritic cell formulation that stimulates NKT cells, from monocytes collected from a recipient (monocytes collected by apheresis or peripheral blood containing monocytes), and that can exhibit the effect more quickly than conventional products. [Solution] This method for producing a dendritic cell formulation comprises: an adhesion step for placing collected monocytes in a culture vessel using a liquid culture medium, and leaving the culture vessel still to cause a portion of the monocytes to adhere to the inner surface of the vessel; a non-adhering cell removal step for removing non-adhering cells including monocytes other than the cells adhering to the inner surface of the culture vessel; a differentiation step for adding a predetermined factor in the culture vessel to cause the monocytes adhering to the inner surface of the culture vessel to differentiate into immature dendritic cells; a pulsing step for performing, with α-galactosylceramide, pulsing in the culture vessel in which the immature dendritic cells are present in a non-adhering state; and an NKT-stimulating dendritic cell induction step for inducing, from the immature dendritic cells, NKT-stimulating dendritic cells that stimulate NKT cells. The dendritic cell formulation is administered back into a recipient from which the blood have been collected.

To construct an oogenesis system not dependent on biological tissue by inducing pluripotent stem cells to differentiate into ovarian somatic tissue.SOLUTION: Differentiation into ovarian somatic tissue can be induced by culturing epiblast-like cells differentiated from pluripotent stem cells in the presence of a BMP agonist and a Wnt agonist, and subsequently culturing in the presence of a BMP agonist, retinoic acid, an SHH agonist, and an FGF inhibitor.SELECTED DRAWING: None

A method for enhancing extracellular vesicle production is disclosed herein.

Provided are a perfusion culture method and the like that are able to improve the productivity in the production of recombinant protein. Specifically, a method for improving recombinant protein production in perfusion culture of an animal cell having a capacity to produce a recombinant protein, the method comprising adding cystine and / or ammonium iron citrate to a cell culture medium, and the like, are provided.

There is provided a culture medium for expansion culture of a mammalian hematopoietic stem cell, where the culture medium contains a glutamate dehydrogenase inhibitor as an active ingredient.

In the present invention, factors attributable to nucleic acids that affect the dynamics of extracellular vesicles are screened.　A library of barcoded extracellular vesicles is provided.

The purpose of the present invention is to provide a novel coating film formation composition for the formation of a coating film having a function of inhibiting adhesion of a biological substance, a coating film using said composition, and manufacturing methods for a substrate for a cell culture, a multilayer film, and a cell aggregate using the coating film.　The present invention provides a coating film formation composition including: a polymer that includes structural units represented by formula (1) and formula (2) (in the formulas, R1-R3, X1, X2, T1, n1, n2, and n3 are as set forth in the specification and the claims) and that does not include a polysiloxane skeleton; and a solvent.

A method of growing isolated mitochondria includes selecting a source of mitochondria and stem cells, extracting the stem cells and the mitochondria from the source and segregating the stem cells and the mitochondria into first and second pools respectively, incubating / transferring the extracted mitochondria into the extracted stem cells to produce packed stem cells, expanding the packed stem cells in a bioreactor, adjusting conditions of an environment of the bioreactor to favor growth of the mitochondria of the packed stem cells, converting the packed stem cells into megakaryocytes, isolating the mitochondria from the megakaryocytes, and applying a coating to the mitochondria following their isolation from the megakaryocytes.

The present invention provides a method for culturing adherent cells, comprising a step for subjecting adherent cells to suspension culture in a medium including nanofibers composed of a water-insoluble polysaccharide, wherein the culture is conducted while stirring.

The present invention provides an acoustofluidic microfluidic chip and system, and a use thereof. The acoustofluidic microfluidic chip comprises a microporous array chip, a microfluidic channel layer, and a bulk acoustic wave resonator array chip which are sequentially stacked in a thickness direction. The acoustofluidic microfluidic chip of the present invention uses acoustofluidics to manipulate micro-objects such as cells or cell secretions, can realize accurate capture, in-situ cell culture, and selective recovery of the micro-objects, has the advantages of simple structure, low cost, high flux, high integration and the like, and has high application value.

Compositions, systems, and kits are disclosed that comprise at least one immune checkpoint inhibitor and at least one immune cell type for adoptive cell therapy, wherein the at least one immune cell type has been fucosylated ex vivo (fuco-ACT). Also disclosed are methods of making and using the compositions, systems, and kits.

According to the mesenchymal stem cell exosome and the preparation method and application thereof provided by the invention, miR-29b-3p with a therapeutic effect is wrapped in the exosome, so that the stability of the miR-29b-3p can be enhanced, the expression of the miR-29b-3p can be effectively regulated, myocardial cell injury is reduced, and myocardial ischemia reperfusion injury is improved; the exosome serving as a carrier can transport miR-29b-3p with a therapeutic effect to myocardial ischemia cells, the targeted delivery efficiency of the miR-29b-3p is improved, systematic side effects are avoided, the therapeutic effect of the exosome and the therapeutic effect of the miR-29b-3p can be combined, the effectiveness of treating myocardial ischemia-reperfusion injury is improved, and in addition, the exosome can be used for preparing medicines for treating myocardial ischemia-reperfusion injury. The ischemic myocardial targeting peptide IMTP is modified on the outer surface of the exosome, so that the targeting property and effectiveness of treatment can be improved.

The invention relates to the technical field of biotechnology and immunotherapy, and discloses a method for inducing and amplifying pMHC specific homologous TSCM, which comprises the following steps: a) preparing pMHC presenting a single antigen peptide; b) sorting lymphocytes and mononuclear cells from a donor, and connecting the pMHC presenting the single antigen peptide obtained in the step a) to the surface of the separated mononuclear cells as stimulating cells; c) co-culturing the sorted lymphocytes serving as effector cells and stimulated cells in a culture medium containing a glycogen synthase kinase-3beta inhibitor, and inducing to generate pMHC specific homogeneous TSCM; and d) separating the pMHC specific homologous TSCM obtained in the step c). The method can induce and amplify sufficient pMHC specific homogeneous TSCM for adoptive immunotherapy, overcomes self tolerance and avoids or alleviates GVHD (Growth Vitamin Horse Disease); the preparation method is simple, induction and amplification efficiency is high, and universality and flexibility are achieved.

The present disclosure provides a method for chemoselective modification of a cell surface molecule on a target cell. A subject method includes contacting a target cell comprising cell surface molecule comprising a thiol, an amine, or an imidazole with a biomolecule comprising a reactive moiety, wherein the reactive moiety is generated by reaction of a biomolecule (e.g., an antibody) comprising a phenol moiety or a catechol with an enzyme capable of oxidizing the phenol or the catechol moiety. The contacting is carried out under conditions sufficient for conjugation of the cell surface molecule to the biomolecule, thereby producing a modified cell. The present disclosure provides kits for carrying out a subject method. The present disclosure also provides modified cells and methods for using same.

Provided is a method for inducing differentiation into nerve cells, the method comprising: a) culturing mesenchymal stem cells for 1-10 days in a stem cell culture medium configured by including a basal medium at 60.0–90.0 vol% and physiological saline at 10.0–40.0 vol%, the culture medium being further supplemented with 1.0–100.0 ng / mL of EGF, 0.2–20.0 ng / mL of FGF-2, 0.2–20.0 ng / mL of PDGF, and 0.5–8.0 mM of magnesium ascorbyl phosphate; b) thereafter, precipitating protein components from the culture solution obtained in step a) using an organic solvent; and c) isolating the precipitate portion and dissolving the same in a physiological saline or a basal medium to obtain a medium comprising a cell-produced protein (CPPs) composition, and seeding, using the medium, the mesenchymal stem cells at a density of 3×103 to 2×104 cells / mL and culturing for 1-3 days, thereby inducing differentiation of the mesenchymal stem cells into nerve cells.

The invention relates to the technical field of biological medicines, and particularly discloses an application composition of stem cell exosomes in promoting tissue repair, which comprises the stem cell exosomes selected from induced pluripotent stem cells, mesenchymal stem cells, embryonic stem cells, hematopoietic stem cells or neural stem cells and used for promoting tissue repair and cell regeneration, and the stem cell exosomes selected from the induced pluripotent stem cells, the mesenchymal stem cells, the embryonic stem cells, the hematopoietic stem cells and the neural stem cells. The tissue repair promoting factor is selected from an epidermal growth factor, a transforming growth factor beta, a platelet-derived growth factor or a combination thereof, and is used for optimizing cell proliferation, differentiation and wound healing; the biodegradable carrier is selected from hydrogel, lipidosome, polymer micelle, nano particles or natural plant extract; by optimizing the transmission relation between the biodegradable carrier and the stem cell exosome and combining with the cell proliferation model, different doses and delivery modes can be flexibly regulated and controlled, the treatment scheme is further optimized, and the effect of the treatment scheme is accurately predicted. Due to the design, the common problems of unstable effect, long treatment period and the like in the traditional stem cell application are solved.