2982results about "Culture process" patented technology

The invention discloses a 3D intestinal organ differentiation method based on human pluripotent stem cells and an induction culture medium and application thereof, and relates to the technical field of stem cells. According to a culture medium formula combination, intestinal organs can be differentiated into various cell types such as epithelial cells, neuroendocrine cells and endothelial cells; the method is a key mark for successful differentiation and functional maturation of intestinal organs. According to the 3D intestinal organ differentiation method disclosed by the invention, histological structures such as intestinal crypts are differentiated from intestinal organs generated by differentiation, and the intestinal organs can creep in a maintenance stage, so that the intestinal organs are changed from structural bionics to functional simulation, and the significance of the 3D intestinal organ differentiation method is far better than that of pure morphological simulation. Through a systematic culture medium formula, a clear operation process and a multi-stage induction strategy, the 3D intestinal organ with structural integrity, cell diversity and functional activity is successfully constructed, and the system provides an efficient, reliable and extensible in-vitro model platform for intestinal biological research and related application.

Disclosed are methods of treating traumatic brain injury in a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

The invention relates to a culture method for activating NKT cells by DC cells in vitro and application of the NKT cells. The preparation method of the NKT cell comprises the following steps: activating a DC cell by using cell inducing factors such as alpha-GalCer, GM-CSF and IL-4, co-culturing the DC cell and a cryopreserved and resuscitated suspension cell left after adherent sorting, and activating and amplifying the NKT cell. The proportion of NKT (CD3 + CD56 +) prepared by the invention is more than 80%, and a small amount of iNKT cells are contained at the same time.

Provided herein are altered cell lines comprising a suspension cell line adapted from an adherent cell line having a different expression profile from a corresponding non-altered adherent cell line, methods for generating altered cell lines, and methods of characterizing altered expression profiles for a gene, a transcript, or a protein in an altered cell line.

A brain organoid containing optic vesicles generated based on H9 induction and an eye-brain fusion culture method thereof are provided. H9 embryonic stem cell induction is used to generate a brain organoid containing optic vesicles with primitive visual field. Based on an established optic vesicle brain organoid culture system, microscopic imaging is combined with specific marker antibodies related to early retinal development and photoreceptor cell maturation to structurally and functionally identify the brain organoid containing optic vesicles.

The invention provides a gamma delta T cell preparation as well as a preparation method and application thereof, and belongs to the technical field of T cells. The liposome is prepared from the following raw materials in parts by weight: 15-20 parts of modified gamma delta T cell liposome, 1-3 parts of cell stimulating factors and 4-7 parts of culture medium freeze-dried powder, the modified gamma delta T cell lipidosome is prepared by embedding gamma delta T cells through lipidosome, coupling with acetylenic bond modified chitosan, and further mixing with MFAP4 protein and b4GALT1 protein. The culture medium freeze-dried powder is prepared by freeze-drying the culture medium in the engineering culture process of gamma delta T cells, and the cell stimulating factors are resveratrol and quercitrin. According to the gamma delta T cell preparation prepared by the invention, the survival ability and resistance of gamma delta T cells are improved, the anti-tumor activity of the gamma delta T cells is improved, and the gamma delta T cell preparation has relatively good antioxidant and anti-inflammatory effects, reduces the inhibition of inflammation on an immune system and has a very good treatment effect on pancreatic ductal adenocarcinoma.

Disclosed are methods of treating perianal fistula a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

The invention discloses a cord blood-derived regulatory T cell amplification culture medium and a use method thereof, and relates to the field of animal cell culture and immune cell therapy, the culture medium is a serum-free culture medium, the culture medium is composed of a basic culture medium, a serum substitute combination, a combination of four cell factors, an immunosuppressor, an apparent stabilizer and an antioxidant, and all the components play a role synergistically; according to the use method, cell amplification is realized through a collaborative process of staged environmental regulation and targeted activation. According to the method, the problems of exogenous pollution and batch difference caused by dependence of a serum-containing system on cord blood-derived regulatory T cell amplification in the prior art are solved, the defect that a serum-free scheme is difficult to consider the amplification efficiency and the function stability at the same time is overcome, the cell amplification quality and the clinical application safety are guaranteed, and the method is suitable for large-scale clinical transformation requirements.

The invention relates to a cell collection method capable of simultaneously collecting three cells in a co-culture model, and belongs to the technical field of biology. The invention provides a cell collection method capable of simultaneously collecting three cells in a co-culture model, and the cell collection method comprises the following steps: after a three-cell co-culture model is constructed, taking out a Transwell chamber, retaining cells in the lower chamber, and collecting the cells in the lower chamber; respectively digesting the cells on the two sides of the Transwell cell membrane by using a trypsin solution with the concentration of 0.5 g / 100mL so as to respectively collect the cells on the two sides of the Transwell cell membrane. According to the cell collection method disclosed by the invention, the three cells in the three-cell co-culture model are simultaneously collected in a manner of digesting the cells on the two sides of the Transwell membrane step by step by using pancreatin, so that not only is the cell and consumable cost saved, but also the experimental synchronism of the three cells is ensured, and convenience is provided for optimizing the experimental process.

The invention belongs to the technical field of immunotherapy, and particularly relates to a method for improving NK cell functions by targeted inhibition of lactylation. The invention provides a method for improving functions of NK cells by targeted inhibition of lactylation, NK cell lactic acid transporters or modification enzymes in inhibition of the lactylation process are targeted, the modification enzymes are NK cell lactylation modification enzymes Writer or lactic acid coenzyme A synthetase, NK92MI lactylation modification is blocked, so that function depletion of the NK cells is reversed, and the NK cell function is improved. The function of the NK cell can be partially reversed by inhibiting the lactic acid modification, and the expression level of effector molecules of the NK92MI cell can be recovered.

The invention relates to methods for culturing cells, and cultured animal cells for use in described method. Also provided is a modified animal cell and guide RNAs targeting genes to be modified in the modified and cultured cells of the invention.

The invention discloses a preparation method of DNA (Deoxyribonucleic Acid) hydrogel with programmable mechanical strength. By regulating and controlling the molar ratio of the catalytic initiating chains to the total amount of the DNA monomer building blocks, multiple hydrogels which are equivalent in mass concentration and have different mechanical strengths are constructed. According to the method, a strategy similar to catalytic assembly is adopted, the feeding molar ratio of a catalytic initiating chain to a DNA monomer building block is creatively adjusted, the proportion of cross-linking reactions at different levels is regulated and controlled, then the DNA hydrogel with different mechanical strengths is synthesized under the isothermal condition, and the storage modulus range is about 200-2000 Pa. In addition, the change of the mechanical strength is independent of the total concentration of the matrix (DNA) of the hydrogel, the DNA sequence, the stress relaxation rate and other properties. The method provides a novel convenient method for constructing DNA hydrogels with different mechanical strengths, and possibly provides a novel tool for researching the specific influence of mechanical properties of materials on cell / tissue behaviors.

The invention relates to the technical field of biological products, and discloses a mesenchymal stem cell hair growth liquid and a preparation method thereof, and the mesenchymal stem cell hair growth liquid comprises the following components: hair follicle mesenchymal stem cells; a hair follicle mesenchymal stem cell exosome; growth factors: a vascular endothelial growth factor (VEGF), a fibroblast growth factor (FGFs), a keratinocyte growth factor (KGF-7) and interleukin-10 (IL-10); vitamins: vitamin E, vitamin B5, vitamin B2, vitamin B6, vitamin C and biotin; the plant extracts comprise a cacumen biotae extract, a polygonum multiflorum extract, a ginseng extract, a peppermint extract, a purslane extract and a rhodiola rosea extract; other active ingredients: tripeptide and digoxin; the auxiliary material is a hyaluronic acid-collagen peptide graft copolymer; and the balance of a phosphate buffer solution.

The present invention relates to a cell biomass (e.g. food product or pet food product) or its components prepared from cultivated metazoan cells, method of its preparation and a system for its preparation.

Disclosed are methods of improving perioral and face fat graft retention in a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

The invention provides a specific sequence nucleic acid molecule and application thereof as a PDRN, and relates to the technical field of biology. A novel nucleic acid molecule with a specific sequence is obtained, and the sequence has certainty, biological activity and stable product quality. The PDRN of the specific sequence does not have cytotoxicity, is high in safety and has a wide application prospect. Moreover, compared with the extracted PDRN, the PDRN with a specific sequence is found for the first time, the product has a remarkable effect of promoting VEGF expression, cell damage can be effectively repaired, and a novel skin-care and repairing functional component is provided for the industries of cosmetics and medicines. The invention further provides a preparation method of the PDRN with the specific sequence, and the preparation method is simple and suitable for popularization, application and industrial production.

The invention relates to the technical field of cell engineering, and particularly discloses a construction method and application of a sika deer immortalized renal epithelial cell line, and the construction method comprises the following steps: taking renal epithelial primary cells from healthy sika deer renal cortex tissues to obtain primary renal epithelial cells; treating the primary renal epithelial cells with trypsin to obtain a cell suspension; inoculating the cell suspension into a complete culture medium, and transfecting with lentivirus loaded with SV40 large T antigen genes to obtain transfected cells; 3 [mu] g / mL puromycin is applied to the transfected cells for selective culture, so that non-transfected cells are eliminated, and drug-resistant cells are obtained; carrying out continuous passage on the drug-resistant cells for at least 30 generations to obtain the immortalized renal epithelial cell line of the sika deer; according to the invention, through a specific action mechanism (inhibiting a p53 / pRb pathway and blocking cell cycle exit) of the SV40 large T antigen, a proliferation limit caused by inhibition of telomerase activity of primary cells of the cervidae animals is overcome, and a cell resource library capable of realizing continuous passage is established.

The invention discloses a bone organ chip loaded with a microelectrode array and a use method, and relates to the technical field of organ chips, the bone organ chip is made of a polydimethylsiloxane material, and the bone organ chip comprises a three-layer structure: a top layer, a middle layer and a bottom layer; the middle layer comprises two blood vessel micro-channels, two osteoblast micro-channels and a neuron micro-channel, the channels are separated through columns and micro-columns, and cell interaction in a bone micro-environment is simulated; a microelectrode array is arranged on the top layer and is used for monitoring electrical activity of neurons in real time; the bottom layer is provided with a pneumatic valve which can control opening and closing of the micro-channel. According to the invention, nerve and blood vessel channels are introduced, a nerve-blood vessel-immunoregulation bone microenvironment is constructed, physiological processes such as osteogenesis-blood vessel formation coupling and the like are embodied, intervals of different sizes can enhance intercellular crosstalk and allow blood vessels and neuron axons to selectively pass through, so that the chip can simulate a complex microenvironment of bone tissues, and the microenvironment of the bone tissues can be simulated; and an efficient and accurate in-vitro model is provided for bone disease mechanism research and drug development.

The invention discloses a preparation method and application of a novel hybrid extracellular vesicle. According to the invention, vesicles derived from endothelial cells and neutrophil cells are combined with deferoxamine, and a biological mixed nano vesicle platform (DFO (HEVS)) is developed to solve the problem of diabetes wound healing. According to the dual-targeting system, DFO is accurately delivered to a wound part by utilizing CXCR4 mediated endothelial cell homing and beta2 integrin dependent inflammation tropism, DFO (at) HEVs activates and recovers vascular regeneration through HIF-1alpha / VEGF, ferroptosis is inhibited through Nrf2 / GPX4 signal transduction, macrophages are reprogrammed into a repair promoting M2 phenotype, and oxidative stress-inflammation-ferroptosis circulation is effectively broken.

The invention relates to a retina nerve and blood vessel unit in-vitro three-culture model and a preparation method thereof, and belongs to the technical field of biology. The invention provides an in-vitro three-culture model of a retina nerve and blood vessel unit. The in-vitro three-culture model is obtained by carrying out in-vitro Transwell indirect co-culture on retina microvascular endothelial cells, retina ganglion cells and retina Muller cells. The in-vitro three-culture model of the retina nerve and blood vessel unit is based on a three-cell indirect co-culture method, retina microvascular endothelial cells, retina ganglion cells and retina Muller cells are simultaneously incorporated by utilizing Transwell, and the three cells are in indirect contact. The in-vitro three-culture model of the retina nerve and blood vessel unit can be used for more objectively simulating pathophysiological states of various cells of the retina nerve and blood vessel unit, so that common pathophysiological changes of mutual dependence and interaction of the three cells under in-vitro conditions can be observed.

The present disclosure relates to methods for the production of a cell-based product based on using synthetic promoters or a cellular input-output unit thereof, configured for controlling the expression of at least one nucleic acid sequence of interest, upon sensing dynamic cellular- and / or environmental- state and / or condition, that leads to a desired phenotype of the cells. The present disclosure further provides cell-based products, in particular, animal cell-based meat products (ACBM).

The invention relates to the technical field of biological medicine, in particular to a cell culture medium and a preparation method of gamma delta T cells. The cell culture medium is a culture medium A or a culture medium B. The culture medium A comprises a serum-free culture medium, 1-100 [mu] M of zoledronic acid, 50-500 U / mL of IL-2, 1-5 [mu] M of imiquimod, 1-20 ng / mL of IL-15, 1-50 ng / mL of IL-1b, 10-100 ng / mL of IL-7 and 2-10% (v / v) of inactivated plasma or a serum substitute. The culture medium B comprises a serum-free culture medium, 100 to 2000 U / mL of IL-2, 1 to 50 ng / mL of IL-15, 1 to 10 [mu] M of zoledronic acid and 2 to 10% (v / v) of inactivated plasma or a serum substitute. By adopting the cell culture medium, a large number of gamma delta T cells can be rapidly obtained.

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 discloses an NKT cell culture optimization method, and belongs to the technical field of cell culture.The method comprises the steps that firstly, PBMC is sorted through a spiral micro-fluidic chip integrated with CD3 / CD56 double-labeled magnetic beads, and CD3 + CD56 + cells with the purity larger than or equal to 95% are obtained through the synergistic effect of dielectrophoretic force and a magnetic field; then, in a GIBCO culture solution added with the IL-2 / IL-15 / alpha-GalCer sustained release microspheres, the pH, dissolved oxygen and lactic acid concentration are accurately regulated and controlled through a real-time monitoring system, and temperature-oxygen partial pressure gradient culture is adopted; and finally, three times of centrifugation are combined with NKT4 solution treatment, and the cells are collected in cooperation with a gradient cooling procedure, so that the cell activity maintenance rate is greater than or equal to 95%. Compared with the traditional technology, the method has the advantages that the NKT cell sorting purity and activity are remarkably improved, the culture environment is regulated and optimized through dynamic metabolism, and accurate control over the culture process is achieved.

The invention discloses a construction method and application of an in-vitro callus-like organ, the survival rate of the organ is improved by improving freeze-formed bio-ink, and a stable directional channel structure is further prepared, so that the internal mass transfer problem of the callus-like organ is improved, and the material exchange efficiency is improved. The forming mode of the callus-like organ is simple and effective, the stent material is easy to prepare, and the callus-like organ has mature cross-linking and printing characteristics, is expected to be applied to drug screening research or bone defect repair, is clear in process and simple in composition, and has relatively high operability.

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.

To provide induced regulatory T cells with high function and stable immunosuppressive function.SOLUTION: The present invention provides an induced human regulatory T cell having at least one characteristic selected from the group consisting of FoxP3-positive, CTLA4-positive, NT5E-positive, ITGAE (CD103)-positive, AREG-positive, CD172 g-positive, and CD26-positive, which is induced by obtaining stable inducible T cells induced from human peripheral T cells by stimulation with an anti-CD3 antibody when regulatory T cells are induced from human peripheral T cells, subsequently subjecting the cells to resting culture, stimulating the cells with anti-CD3 antibody again to culture them, and further subjecting the cells to resting culture.SELECTED DRAWING: None

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.