18results about How to "High differentiation efficiency" patented technology

The invention discloses a method for differentiating induced pluripotent stem cells into macrophages. The method comprises the following steps: S1, inducing pluripotent stem cells to form embryoid bodies; s2, differentiating the embryoid body into a monocyte with CD14 expression; s3, the mononuclear cells are differentiated into macrophages with CD14 / CD11b expression; and S4, polarizing the macrophages into M1 type macrophages and / or M2 type macrophages. According to the method for differentiating the induced pluripotent stem cells into the macrophages, the cells can be effectively induced to differentiate towards the macrophages, the induced pluripotent stem cells are subjected to directional differentiation culture in a short time to obtain the macrophages, and the generated macrophages express CD14 and CD11b and have remarkable phagocytic activity on yeast glycan. The method is of great significance to large-scale production of macrophages.

The invention relates to the field of cell biology, and particularly discloses a neural stem cell induced differentiation culture medium and application thereof in preparation of neural stem cells. The neural stem cell induced differentiation culture medium is composed of a basic culture medium and additives, wherein the additives are composed of an N-2 additive, a B-27 additive, glutamine, a human leukemia inhibiting factor, a GSK-3 inhibitor, an ALK inhibitor, a gamma-secretase inhibitor and a CDK8 inhibitor. The neural stem cell induced differentiation culture medium is clear and simple in component, does not need to use antibiotics, and does not contain animal-derived components. When the culture medium is used for carrying out differentiation induction on the human pluripotent stem cells, the human pluripotent stem cells can be induced and differentiated into the neural stem cells within a short time (6-7 days) without depending on pluripotent stem cell cloning and a single cell state, the nonuniformity of pluripotent stem cell cloning formation can be overcome, and the purity of the neural stem cells is improved.

The invention discloses a yak intramuscular precursor fat immortalized cell line and a preparation method thereof. The method comprises the following steps: (1) obtaining yak primary intramuscular precursor fat cells; (2) performing SV40T lentivirus infection on the precursor fat cells in the yak muscle and screening cell strains: performing SV40T lentivirus infection by utilizing a 1 / 2 small-volume infection method to obtain immortalized cells after the precursor fat cells in the yak primary muscle are infected with SV40T, namely immortalized cells for short; puromycin is adopted to screen immortalized cells; and diluting the screened cells by using a limited dilution method, and carrying out enlarged culture to obtain the immortalized cell line. The yak intramuscular precursor fat immortalized cell line constructed by the invention has genetic homogeneity, the heterogeneity influence of cell populations is eliminated, and an in-vitro cell model is provided for subsequent research related to yak intramuscular fat cells.

The invention belongs to the technical field of plant tissue culture, and particularly relates to a method for inducing Hangzhou chrysanthemum leaf adventitious bud regeneration by using a potato extract, which sequentially comprises the following steps: cultivating Hangzhou chrysanthemum stem segments with buds into Hangzhou chrysanthemum aseptic seedlings with at least four leaves; the method comprises the following steps: taking down 3-4 leaves from the top of a sterile seedling of Hangzhou chrysanthemum, taking a leaf disc from each leaf close to a petiole by using a puncher, immediately inoculating the leaf disc onto an adventitious bud induction culture medium, and culturing in a plant tissue culture room; and obtaining Hangzhou chrysanthemum leaf adventitious buds. According to the method, the leaf disc adventitious bud induction time is short, the growth state is good, subsequent normal rooting, transplanting and breeding can be carried out, and a foundation is laid for successfully establishing a Hangzhou chrysanthemum genetic transformation system.

The invention discloses a co-culture method of cell culture meat and application of the co-culture method in improving meat quality and flavor. According to the invention, through two-stage differentiation, muscle precursor cells and fat precursor cells are respectively subjected to proliferation culture, then are separately subjected to primary differentiation, and are mixed together for co-culture, and meanwhile, second-stage continuous differentiation is carried out; the problem that differentiation of muscle cells and fat cells cannot be effectively and simultaneously differentiated under the co-culture condition due to different requirements of differentiation of muscle cells and fat cells on the serum environment is solved. In addition, the invention also provides an objective standard for evaluating the flavor quality of cell culture meat generated by co-culture. The cell culture meat with different flavors can be obtained by adjusting the proportion of the co-culture cells through the co-culture method, and the flavor quality of the cell culture meat from other sources can be evaluated and detected through the amino acid detection means.

The invention discloses a vascular endothelial cell as well as a preparation method and application thereof. The preparation method comprises the following steps: inoculating induced pluripotent stem cells into a culture medium containing an ROCK signal channel inhibitor for culturing, and inducing the cultured induced pluripotent stem cells by using a culture medium containing a Wnt signal channel activator and a BMP signal channel activator, so that the induced pluripotent stem cells are differentiated into mesoderm cells; inducing the mesoderm cells by using a culture medium containing a vascular endothelial growth factor and a cAMP signaling pathway agonist, so that the mesoderm cells are differentiated into endothelial coarse cells; and performing positive sorting of the endothelial cell marker CD144 on the endothelial coarse cells to obtain purified vascular endothelial cells. The induction path of the method is clear and efficient, the period is short, the vascular endothelial cell population with high purity, complete vascular barrier function and mature phenotype can be obtained, and the cell population can be applied to construction of a vascular network in a gel matrix.

This invention discloses the application of sorafenib in the preparation of drugs for treating lung metastases, relating to the field of biomedical technology. This invention focuses on CD4... + Sorafenib treatment of CD4 cells during T cell differentiation into Th9 cells + T cells were found to significantly improve the differentiation efficiency of Th9 cells in vitro, indicating that sorafenib can expand Th9 cells with anti-tumor effects in large quantities in vitro, providing a new idea and approach for ACT therapy in clinical tumors.

The invention relates to an application of citric acid in promoting chondrogenesis induced differentiation of adipose-derived stem cells, a culture solution for promoting chondrogenesis induced differentiation of adipose-derived stem cells and a method for promoting chondrogenesis induced differentiation of adipose-derived stem cells, and the method comprises the following steps: culturing adipose-derived stem cells in a culture medium for promoting chondrogenesis induced differentiation of adipose-derived stem cells, bone differentiation cells are obtained, and the bone differentiation cells are chondroblasts; the culture medium takes a chondrogenesis induction culture medium as a basic culture medium, the culture medium further comprises citric acid, and the concentration of the citric acid is 0.5-2mM. The culture solution containing citric acid is used for culturing the adipose-derived stem cells, so that the cells can obtain sufficient nutrition, cell proliferation and differentiation are facilitated, the biocompatibility is good, degradation is achieved, the cell survival time is prolonged, the cell morphology is maintained, the proliferation capacity of the adipose-derived stem cells is improved, the expression quantity of specific genes of cartilage cells is up-regulated, and the differentiation efficiency is improved.

The application discloses TaHAT1 The application belongs to the technical field of plant genetic engineering and relates to application of a gene in promoting differentiation efficiency and genetic transformation of wheat callus. TaHAT1 The application discloses that overexpression of the gene in wheat young embryo explants can improve the differentiation efficiency and genetic transformation efficiency of wheat callus, and the growth and development of the transformed plants are normal, which provides a new genetic transformation gene resource for biological breeding of wheat and has important significance for promoting genetic improvement of wheat.

The present invention discloses an in vitro treatment method for improving the differentiation efficiency of Th9 cells, and relates to the field of biomedicine technology. + When T cells differentiate into Th9 cells, sorafenib is used to treat CD4 + T cells, and found that sorafenib can significantly improve the differentiation efficiency of Th9 cells in vitro, indicating that sorafenib can amplify Th9 cells with anti-tumor effects in vitro, providing new ideas and new approaches for the ACT treatment of clinical tumors.

The application belongs to the technical field of regenerative medicine, and discloses a kind of organoid culture method derived from human and non-human primate subventricular zone neural stem cells. By optimizing the medium formula and regulating transcription factors, the organoid is successfully induced to differentiate into CRABP1-expressing neuronal precursors. These cells express TAC3, and the application overcomes the limitations of existing technologies in simulating primate-specific neural lineages and complex brain structures, providing a breakthrough in vitro model for studying neural development, neurological diseases and drug development. The established system not only provides a new tool for studying the development of primate nervous system and disease mechanism, but also provides a reliable experimental platform and transplantable cell source for developing targeted drugs, constructing advanced disease models and conducting cell replacement therapy.

The invention discloses application of sorafenib in preparation of a medicine for resisting lung metastatic tumors, and relates to the technical field of biological medicines. When the CD4 + T cells are differentiated to the Th9 cells, the sorafenib is used for treating the CD4 + T cells, so that the in-vitro Th9 cell differentiation efficiency can be remarkably improved by the sorafenib, the Th9 cells with the anti-tumor effect can be massively amplified in vitro by the sorafenib, and a new thought and a new way are provided for an ACT treatment method of clinical tumors.

The application relates to the technical field of genetic engineering, in particular to a nucleotide sequence for improving the differentiation efficiency of Picea abies callus, an expression vector and application thereof, which comprises a coding sequence of an AP2L2 gene, and the expression vector comprises double-strand RNA composed of the nucleotide sequence and its reverse complementary sequence. The application discloses the negative regulation of the AP2L2 gene in the differentiation of Picea abies callus for the first time, and effectively solves the problem of reduced differentiation efficiency by using specific gene silencing technology. The scheme is feasible and has remarkable effect, and is suitable for large-scale breeding production of forest tree varieties.

The invention relates to the technical field of stem cell culture, in particular to an AI algorithm for evaluating the growth state of mesenchymal stem cells, which comprises the following steps: acquiring mesenchymal stem cell image data, and inputting related basic information; extracting morphological characteristic data of the cells from the image data by utilizing a stem cell culture technology and a deep learning algorithm, and judging whether the cells have pollution, degeneration or abnormal morphology; based on the cell morphological characteristic data, analyzing the growth stage of the cells, and adjusting culture conditions according to the characteristics of the stage; collecting epigenetics data of stem cells, predicting cell proliferation and differentiation trends by using a deep learning model in combination with cell morphological characteristic data and cell growth stage analysis results, and adjusting culture conditions; and monitoring metabolite data in the stem cell culture solution in real time, evaluating the health state of the cells by combining the metabolite data, the cell morphological characteristic data and the cell cycle analysis result, and judging whether the problem of abnormal metabolism or decline exists or not.

The application discloses a preparation method of a drug sustained-release hydrogel support and a three-dimensional osteogenic induction application of hiPSCs. Firstly, a CHIR99021@BSA-DEX@PLGA time-sequential double sustained-release composite microsphere of a core-shell structure is prepared, then the freeze-dried GelMA powder is dissolved in a PBS solution containing a photoinitiator LAP, the CHIR99021@BSA-DEX@PLGA time-sequential double sustained-release composite microsphere is added after stirring and cooling, and a drug-loaded hydrogel precursor is obtained; the precursor is injected into a mold and irradiated by a specific wavelength light source to initiate free radical polymerization of a methacryl group into glue, and a drug-loaded GelMA porous hydrogel support is obtained after demolding, freezing and drying; the drug-loaded GelMA porous support is covalently grafted with VN polypeptide through EDC / NHS chemical coupling to obtain a VN-GelMA drug-loaded porous hydrogel support. The application simulates the signal time sequence regulation law in the embryonic bone formation process and significantly improves the osteogenic differentiation efficiency and mineralization quality by constructing a biomimetic three-dimensional osteogenic microenvironment. The VN polypeptide enhances the specific recognition of stem cells and significantly improves the three-dimensional cell compatibility. The application can reduce the dependence on exogenous induction factors and is safe, low in cost and stable in induction.

The application discloses a reagent for preparing macrophages by human pluripotent stem cell differentiation and application thereof. Specifically disclosed are a reagent and a method for preparing macrophages, wherein the reagent comprises a culture solution and a 3D micro-scaffold material, and the culture solution is composed of culture solution II, culture solution III, culture solution IV, culture solution V and culture solution VI. The application utilizes the 3D micro-scaffold to develop a chemical composition-determined, animal-derived component-free and low-cost stem cell differentiation culture system which is beneficial to clinical-level stem cell differentiation, and can make human pluripotent stem cells continuously and efficiently differentiate into a large amount of high-purity macrophages. The reagent and the method have the characteristics of short time consumption, high differentiation efficiency, low cost and being more beneficial to large-scale automatic industrial production. The preparation method provided by the application can be used for large-scale production of human macrophages, and the quality is stable and the safety is high, thereby providing a large amount of cell sources for tissue engineering, drug research and development and cell therapy.

The invention discloses a culture method of DC-CTL cells, and particularly relates to the technical field of biotechnology and medicine. The invention provides a novel DC-CTL cell culture method based on iPSC, aiming at the technical problems that a traditional DC-CTL cell culture method is limited in cell source, greatly influenced by individual difference, poor in stability and repeatability, insufficient in cell activity and killing ability and the like. A self-made additive and a specific cell factor are combined, SOCS1-siRNA is transfected through a liposome transfection method, the SOCS1 gene is effectively silenced, and the T cell activity is enhanced. According to the method, the proportion of CD3 + CD8 + T cells in the DC-CTL cells reaches 85% or above, the content of IFN-gamma is larger than or equal to 500 pg / mL, the content of TNF-alpha is larger than or equal to 300 pg / mL, and the killing rates under the effect target ratios of 10: 1, 20: 1 and 40: 1 reach 57%, 63% and 72% respectively.

The invention provides a method for forming a regenerated plant by a direct somatic embryogenesis pathway of a pearly konjac, collecting the germination of unopened spathe flames of a pearly konjac class yellow konjac, using tender female inflorescences as explants, screening and optimizing the female inflorescences of the pearly konjac to induce direct somatic embryogenesis, maturation and plant regeneration conditions, and preliminarily establishing an efficient plant regeneration system. Under suitable temperature, light and culture conditions, the somatic embryo directly germinates into seedlings and transplants well. The invention provides a theoretical basis and technical methods for the in vitro preservation of germplasm resources of pearly konjac in Hainan hot zone, rapid breeding of seedlings and large-scale promotion, and the construction of a genetic transformation receptor system regeneration system of pearly konjac, which is of great significance to the sustainable development of the pearly konjac industry in Hainan hot zone.