313results about "Cell membrane modification" patented technology

Provided are a γδT cell targeting PD-L1, and a preparation method therefor and the use thereof. By means of unnatural sugar metabolic labeling, the γδT cell is modified with a first active group, and then same and a PD-L1 targeting group modified with a second active group are subjected to a bioorthogonal reaction, thereby obtaining the γδT cell targeting PD-L1. The γδT cell targeting PD-L1 has a targeted and selective killing effect on PD-L1-positive tumor cells; has the advantages of a short preparation cycle, suitability for large-scale production, uniform and stable quality, etc.; and has good application prospects.

A method for domesticating CHO-K1 cells into high-expression cells, comprising the following steps: culturing adherent CHO-K1 cells in a complete culture medium, to obtain log growth CHO-K1 cells; transferring the log growth CHO-K1 cells to a serum-free medium for suspension culture; performing a first liquid replacement domestication, to obtain domesticated CHO-K1 cells; placing the domesticated CHO-K1 cells in a chemically defined medium for suspension culture; performing a second liquid replacement domestication, to obtain domesticated suspended CHO-K1 cells; performing monoclonalization on the domesticated suspended CHO-K1 cells, to obtain CHO-K1 monoclonal cells; and screening the CHO-K1 monoclonal cells, to obtain high-expression CHO-K1 monoclonal cells. This method can domesticate CHO-K1 original parent cells into suspended cells, and screen cells having higher expression capability from the domesticated cell population, thereby improving expression levels.

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 discloses a GSH response type cell membrane bionic nano-drug, a preparation method and application, the cell membrane bionic nano-drug comprises cell membrane vesicles serving as a carrier, the cell membrane vesicles are prepared by performing cell membrane extraction and membrane extrusion on immune cells of which the membrane surfaces overexpress azide, and the cell membrane vesicles serve as a carrier. The azide on the cell membrane vesicles and the bio-coupling compound are bonded through a click chemical reaction to form-SH groups, and the-SH groups and the small-molecule inhibitor are combined together in a disulfide bond mode through the cross-linking agent. The drug is connected with a small-molecule inhibitor through a disulfide bond, so that the drug loading rate is relatively higher, the stability is better, and drug leakage in the blood circulation process is relatively less; the small-molecule inhibitor can be completely released only in a tumor site microenvironment, so that the tumor m6A methylation level is reduced, the EMT state is reversed, the tumor drug resistance is overcome, and a better tumor treatment effect is displayed in cooperation with chemotherapy drugs for tumor treatment.

The invention discloses a chemical labeling system (HIM-IP) for detecting interaction between cells. According to the system, tyrosinase and phenol are respectively modified on the surfaces of two to-be-detected cells to construct capturing cells and prey cells, and then biotin or single-stranded DNA with nucleophilic groups is used as a molecular tag for labeling. The system not only can perform detection through traditional flow cytometry, but also can be compatible with a commercialized single cell transcriptome sequencing platform to realize association of intercellular interaction information and transcriptome information on a single cell level. The system does not need to carry out complex gene engineering modification on cells, the whole modification and marking process is simple, convenient and efficient, the detection sensitivity of intercellular interaction is high, and the biological mechanism of intercellular interaction regulation and control can be deeply excavated by integrating the single cell sequencing technology.

The present invention provides, in certain aspects, a natural killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15), and methods for producing such cells. The invention further provides methods of using a natural killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15) to treat cancer in a subject or to enhance expansion and / or survival of NK cells.

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 provides a resveratrol-loaded exosome and a preparation method thereof, and belongs to the technical field of biological medicine manufacturing. The preparation method of the resveratrol-loaded exosome comprises the following steps: performing 3D culture on human umbilical cord or hair follicle-derived mesenchymal stem cells under a low-oxygen condition, collecting supernate, and separating by a differential ultracentrifugation method to obtain the exosome of the mesenchymal stem cells; the method comprises the following steps: mixing a resveratrol solution containing a surfactant with the MSCs-Exos, and carrying out co-incubation and ultrafiltration to obtain the resveratrol loaded MSCs-Exos. According to the invention, the surfactant is added, so that not only are the entrapment rate and the drug loading rate of the MSCs-Exos loaded resveratrol greatly improved, but also the activity of the MSCs-Exos is maintained, the common effect of the MSCs-Exos and the resveratrol is fully exerted, and a synergistic effect is achieved. The resveratrol loaded MSCs-Exos disclosed by the invention has an effect of treating diabetes mellitus.

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 discloses miR-150-3p-loaded engineered extracellular vesicles with the surfaces modified with iRGD targeting peptides and application of the miR-150-3p-loaded engineered extracellular vesicles in melanoma treatment, and belongs to the technical field of biological medicines.The miR-150-3p is loaded in EVs, the surfaces of the miR-150-3p are modified with the iRGD targeting peptides, and iEV-150 is obtained; the preparation method of the iEV-150 comprises the following steps: S1, screening miR-150-3p; s2, constructing an HEK293T cell line capable of stably expressing the miR-150-3p; s3, introducing an ANXA2 protein; s4, culturing an HEK293T cell line; s5, separating the EVs; s6, coupling the iRGD targeting peptide to the surface of the extracellular vesicle membrane; the iEV-150 can effectively deliver miR-150-3p to melanoma cells, inhibit tumor cell proliferation, DNA damage repair and epithelial-mesenchymal transition, and promote melanoma cell death by inducing ferroptosis.

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 invention provides a device and a method for realizing particle modification, the device comprises a first flow channel and an acoustic resonator capable of acting on liquid in the first flow channel, the acoustic resonator is fusiform, and the acoustic resonator is used for generating an acoustic fluid effect in the liquid in the first flow channel to capture first particles in the first liquid, the captured first particles are distributed and flow along the fusiform profile of the acoustic resonator, so that the first particles distributed to the side edge of the fusiform profile are controllably mixed with the second liquid in the side edge area, and the controllable mixing is used for controllably combining the second particles in the second liquid with the first particles; the first particles combined with the second particles flow to a fourth flow channel along the tip of the rear end of the fusiform profile of the acoustic resonator; and the first liquid flowing through the first flow channel flows to the two fifth flow channels.

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.

A peptide fragment disclosed herein comprises any of the following amino acid sequences: YLIQFKKRTLRKSNRKKR (SEQ ID NO: 1), FQILYKKRTLRKSNRKKR (SEQ ID NO: 2), YQFLIKKRTLRKSNRKKR (SEQ ID NO: 3), and ILFQYKKRTLRKSNRKKR (SEQ ID NO: 4).

[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

The invention discloses an engineered cell composition as well as a preparation method and application thereof. The cell composition disclosed by the invention is macrophages or mononuclear cells. The preparation method of the composition comprises the following steps: constructing nanoparticles / microparticles, introducing cell specific recognition molecules to obtain specific nanoparticles / microparticles capable of being recognized by macrophages / monocytes, and co-incubating the specific nanoparticles / microparticles and the macrophages / monocytes to prepare the cell composition. The cell composition disclosed by the invention is simple and convenient in preparation method, is easy to carry drugs and probes, and can realize targeted delivery and diagnosis and treatment of infected or tumor parts.

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.