71 results about "Embryonic stem cell" patented technology

The present invention relates to the new use of an exosome secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and in particular relates to the use of the exosome in the preparation of a drug for treating liver diseases. The exosome is secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and the hepatocyte lineage cells are hepatic progenitor cells and / or mature hepatocyte-like cells.

The generation of complex organ tissues from human embryonic and pluripotent stem cells (PSCs) remains a major challenge for translational studies. It is shown that PSCs can be directed to differentiate into intestinal tissue in vitro by modulating the combinatorial activities of several signaling pathways in a step-wise fashion, effectively recapitulating in vivo fetal intestinal development. The resulting intestinal “organoids” were three-dimensional structures consisting of a polarized, columnar epithelium surrounded by mesenchyme that included a smooth muscle-like layer. The epithelium was patterned into crypt-like SOX9-positive proliferative zones and villus-like structures with all of the major functional cell types of the intestine. The culture system is used to demonstrate that expression of NEUROG3, a pro-endocrine transcription factor mutated in enteric anendocrinosis is sufficient to promote differentiation towards the enteroendocrine cell lineage. In conclusion, PSC-derived human intestinal tissue should allow for unprecedented studies of human intestinal development, homeostasis and disease.

This invention discloses the application of alismazone B acetate in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy and its related symptoms. This invention is the first to propose the use of alismazone B acetate (AB23a) for the prevention or treatment of hypertrophic cardiomyopathy. The invention utilizes AB23a to directly treat cardiomyocytes obtained from directed differentiation of human embryonic stem cells in vitro, finding that AB23a treatment significantly inhibits the hypertrophic phenotype of human embryonic stem cell-cardiomyocytes. Feeding mice with hereditary hypertrophic cardiomyopathy caused by gene mutations to a diet containing AB23a showed that AB23a significantly alleviated pathological myocardial hypertrophy and improved cardiac function in mice. AB23a can be used to prepare drugs against hereditary hypertrophic cardiomyopathy, providing a new approach and method for treating hypertrophic cardiomyopathy. AB23a is the most important medicinal component of the traditional Chinese medicine Alisma plantago-aquatica, is safe for organisms, and has good clinical application prospects.

The invention relates to a method of differentiating pancreatic bi-potent progenitor cells into beta cells comprising cultivating pancreatic endocrine precursor cells in a medium that promotes the cAMP signalling pathway that lies downstream of apical-basal polarity in endocrine precursors, which can be used to increase the differentiation of progenitor cells into beta cells. Furthermore, the present invention relates to a method of enriching and / or isolating one or more cells expressing a marker of apical-basal polarity, thereby isolating beta cells, and to a method of producing a population of beta cells from a population of human embryonic stem cells. Furthermore, the present invention relates to a population of beta cells obtainable by the method, a pharmaceutical composition comprising the beta cells, a cell culture of progenitor cells in a medium comprising a cAMP agonist, a complex comprising a beta cell bound by the binding agents specific for CD133 and CD49a, and a use of specific binding partners for isolating beta cells.

The invention discloses a cell culture medium supplement as well as a preparation method and application thereof, and relates to the technical field of cell biology. The supplement comprises DL-alpha-tocopheryl acetate, linolenic acid, ethanolamine, L-carnitine hydrochloride, 3, 3 ', 5-triiodoL-sodium thyrosinate, and serum albumin. As a cell culture additive with definite chemical components and simple composition, the cell culture additive can replace a traditional culture system with unknown chemical components or too complex composition, and is used for supporting proliferation, differentiation and characterization analysis of cells. The culture medium additive is clear in chemical component, simple to prepare, stable in effect and relatively low in cost, can promote in-vitro proliferation and oxidation resistance of cells including human embryonic stem cells and mesenchymal stem cells, and has important application value in actual production.

The invention provides a skin organoid construction method of human embryonic stem cells based on high-expression recombinant human COL17A1 protein and application of the skin organoid construction method, and relates to the technical field of molecular biology, and the skin organoid construction method comprises the following steps: integrating nucleic acid molecules for coding COL17A1 protein to a KRT5 site of genome DNA of the human embryonic stem cells to obtain recombinant human embryonic stem cells; the nucleic acid molecule for coding the COL17A1 protein contains the gene for coding the COL17A1 protein, and compared with the traditional'random integration 'or'universal safe port site' (AAVS1), the yield of the recombinant human COL17A1 protein is remarkably improved by integrating at the KRT5 site, and the stable and efficient expression of the recombinant human COL17A1 protein is realized. The technical problem that in the prior art, the expression quantity of recombinant human COL17A1 protein is low when mammalian cells such as HEK-293T and CHO are used for expressing the recombinant human COL17A1 protein is solved.

The invention discloses a composite elicitor for promoting taxus chinensis embryogenic stem cells to produce paclitaxel and application, and belongs to the technical field of biology. The compound elicitor consists of methyl jasmonate, indolebutyric acid and salicylic acid, and the working concentrations of the methyl jasmonate, the indolebutyric acid and the salicylic acid in a culture medium are respectively as follows: 50-800 mu mol / L of methyl jasmonate, 12.5-50 mu mol / L of indolebutyric acid and 0.1-0.5 mg / L of salicylic acid. Preferably, the jasmonic acid methyl ester is 200 [mu] mol / L, the indolebutyric acid is 25 [mu] mol / L, the salicylic acid is 0.5 mg / L, and 0.1-0.5 mg / L arachidonic acid can be selectively added. The composite elicitor is added in the early stage of the exponential growth phase of the taxus chinensis embryogenic stem cells, and the taxus chinensis embryogenic stem cells are continuously cultured for 5 days, so that the yield of paclitaxel reaches 27.52 mg / L and is 7.86 times that of blank control, and meanwhile, the maximum biomass is obtained. According to the invention, the synergistic interaction of the three elicitors is realized for the first time, the production period is remarkably shortened, a set of efficient and stable paclitaxel production system is established, and an innovative solution is provided for large-scale production of paclitaxel.

This invention provides a device and method for real-time monitoring of stem cell exosome therapy based on a cardiomyocyte sensing system, belonging to the field of micro-nano sensing and exosome therapy monitoring technology. The device includes a cell electrophysiological sensing element 1, an electrophysiological signal acquisition module 2, a signal amplification module 3, a signal display module 4, and a cell culture incubator 5. These components are sequentially connected via cables and data lines, forming a complete signal pathway. This device is used to detect the process of embryonic stem cell exosome therapy. Through multi-concentration comparison and continuous monitoring at multiple time points, it realizes the development and utilization of a new method for monitoring embryonic stem cell exosome therapy. The detection process is real-time, non-invasive, high-throughput, and highly sensitive; the detection method is efficient and simple, and has promising applications in clinical diagnosis and treatment.

This invention belongs to the field of organoid culture technology, specifically involving BCL2 Application of genes in 3D cluster culture of stem cell-derived organoids. Starting on day 4 of bovine embryonic stem cell cluster culture, the cell cluster edges became indistinct, cells began to die and darken, and the cell clusters began to diffuse. By day 7, the cell cluster survival rate was only 14.2%. This invention utilizes the overexpression of genes in bovine embryonic stem cells... BCL2 Following gene expression, the cells were cultured in clusters using the same method. The overexpressed cell clusters showed clear edges, and their volume increased continuously with the number of culture days. Even on day 7, clear cell cluster edges were still observable, and no diffuse cell death was observed; the survival rate remained high at 85.54%. BCL2 Genes can improve the survival rate of early cultured cell clusters of organoids, which is of great significance for the normal morphogenesis and functional maturation of organoids and can greatly improve the efficiency of organoid preparation.

This application discloses a smart method for preparing gene-edited mammalian cells. The method includes: acquiring the target gene sequence information of the mammal to be observed and processing it according to preset screening rules to obtain the target sgRNA sequence; mixing an RNP complex suspension prepared based on the target sgRNA sequence with a suspension of embryonic stem cells to be edited, and performing electroporation transfection using multiple consecutive micropulses to obtain a transfected cell suspension; acquiring raw images of the transfected cell suspension and preprocessing them to obtain single-cell images, and using a trained lightweight convolutional neural network model to obtain the predicted gene-editing efficiency and prediction confidence after 48 hours; if the transfected cell suspension meets the quality control conditions, expanding and culturing the transfected cell suspension to obtain the prepared gene-edited cells. This method can shorten the cell culture cycle, reduce the waste of cell materials, improve the stability of transfection efficiency, and achieve precise preparation of gene-edited cells.

It was discovered herein that neither the nuclear localization signal (NLS) nor the prion-like domain (PLD) of TDP-43 are required for in vitro embryonic stem cell culture and differentiation into motor neurons. ES cells expressing these TDP-43 mutants and differentiated into motor neurons that exhibit an ALS-like phenotype, from which the TDP-43 mutants redistribute to and accumulate in the cytoplasm, and the inability to regulate cryptic exon splicing, such that these cells can serve as a model for TDP-43 proteinopathies for testing candidate therapeutics that can dissipate such proteinopathies. In addition, these ES cells can be used to successfully generate non-human animals, e.g., mice, that also exhibit hallmark symptoms of ALS and can be used to test candidate agents useful for treating TDP-43 proteinopathies.

This invention provides a human embryonic stem cell expansion culture medium, comprising DMEM / F12 medium, ITS-A, HEPES, magnesium ascorbate phosphate, non-essential amino acids, bFGF, FGF2-G3, GlutaMAX, LY-333531, gentamicin sulfate, Y27632, and CHIR99021. Y-27632 inhibits pluripotent stem cell apoptosis, improves adhesion efficiency, maintains pluripotency and colony formation ability; CHIR99021 enhances the self-renewal of pluripotent stem cells and inhibits their differentiation; LY-333531 inhibits spontaneous differentiation of pluripotent stem cells; and gentamicin sulfate reduces the risk of contamination during culture. The culture medium provided by this invention is serum-free, free of allogeneic substances, has a simple composition, and offers excellent expansion speed, effectively maintaining long-term stable passage of human embryonic stem cells, providing stable technical support for scientific research.

The present application relates to a preparation method and application of photoreceptor precursor cells of human embryonic stem cell-derived retinal organoids, and belongs to the field of biotechnology and stem cell engineering. The present application constructs a lentiviral vector carrying an exogenous target gene and infects human embryonic stem cells to obtain a stable expression of engineered stem cell clone group by screening; the clone group is subjected to three-dimensional differentiation, and sequentially undergoes the formation of a blastoid, neural epithelial induction and retinal layering stage to obtain a retinal organoid; finally, the co-expression of photoreceptor precursor cell markers and exogenous genes in the organoid is identified. The present application realizes stable integration of genes at the source of stem cells, avoids the problem of low infection efficiency in the later stage, and provides a complete standardized scheme from gene modification, directional differentiation to function verification. The engineered photoreceptor precursor cells obtained by the method have important application value in the construction of retinal disease models, drug screening and cell therapy development.

This disclosure provides ungulate embryonic stem cells (ESCs) derived from the inner cell mass of pre-implantation blastocysts or pluripotent cells from embryos. From an agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genetic engineering, and providing an experimental tool for studying human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors.

A cell culture method, using a cell culture medium, excluding human embryonic stem cells, the method comprising: (i) contacting the culture medium with at least one cell; (ii) placing the culture medium in a chamber (40) containing a culture medium; the method being characterized in that the medium comprises particles sensitive to a magnetic field, and that the medium has, along a longitudinal axis, a dimension greater than the dimensions along another axis, the larger dimension forming a length of the medium, the method comprising: (iii) applying a magnetic field in the chamber, the magnetic field forming field lines extending through the chamber; (iv) following (iii) suspending each culture medium in a culture medium contained in a chamber. Figure 6

The present disclosure provides methods of manufacturing cell-laden scaffolds comprising lipid-producing cells comprising confining lipid-producing precursor cells in a three-dimensional (3D) environment and culturing the lipid- producing precursor cells confined in the 3D environment in a food-grade differentiation medium to obtain the cell-laden scaffold comprising lipid-producing cells. In one embodiment, the 3D environment is formed by a mixture comprising lipid-producing precursor cells and a food-grade polymericsolution, wherein the food-grade polymeric solution comprises one or more selected from the group consisting of alginate, gelatin, pectin, agarose, carrageenan, gellan gum, konjac glucomannan, cellulose, etc. and combinations thereof, the lipid-producing precursor cells are selected from the group consisting of adipose-derived stem cells, preadipocytes mesenchymal stem cells, embryonic stem cells, induced- pluripotent stem cells and combinations thereof, the lipid-producing cells are selected from adipocytes, hepatocytes, sebocytes, mammary epithelia cells, keratinocytes and combinations thereof, and the food-grade differentiation medium comprises a fatty acid.

The invention discloses a stem cell microcapsule-entrapped tissue adhesive, a preparation method and application, the tissue adhesive is composed of stem cell microcapsule-entrapped hydrogel and a biological adhesive, and live stem cells are entrapped in the tissue adhesive; the stem cells are selected from at least one of umbilical cord mesenchymal stem cells, bone marrow mesenchymal stem cells, adipose-derived stem cells, embryonic stem cells and induced pluripotent stem cells; the biological adhesive is formed by compounding a hydrophilic polymer and a block polymer, wherein the mass ratio of the hydrophilic polymer to the block polymer is (1: 5)-(5: 1); the transmittance of the tissue adhesive is 60-100%, the tissue adhesive strength is 20-200 kPa, and the volume swelling ratio is smaller than or equal to 50%. According to the scheme, the stem cell entrapped microcapsule and the biological adhesive are mixed to prepare the tissue adhesive, and the adhesive can be used as bio-ink to be assembled into a bionic scaffold through 3D printing or directly injected to an injured part, so that the tissue adhesive has a wide application prospect.

The invention provides an embryonic stem cell differentiation technology and application of the embryonic stem cell differentiation technology in screening of unknown risk alternative toxicity of a novel biological breeding product, and Cry1Ab protein is used as a test substance to evaluate the developmental toxicity of the Cry1Ab protein. The EBs are generated through hanging drop culture, and bone differentiation induction substances (beta-glycerophosphate, ascorbic acid and vitamin D3) are added to promote differentiation of the EBs. After bone cell induced differentiation is finished, the differentiation condition of bone cells is observed through alizarin red S dyeing and absorbance value detection, the cell growth condition is observed through cell total protein concentration and alkaline phosphatase activity detection, and then the influence of a test substance on the bone differentiation process is analyzed through the gene expression condition of osteogenic differentiation related markers (Runx2, SPARC and I-type collagen). The invention solves the problem of evaluating the unknown risk of a novel biological breeding product in vitro, creates an embryonic stem cell bone differentiation test technology, and uses the embryonic stem cell bone differentiation test technology as a screening method for the unknown risk alternative toxicity of the novel biological breeding product to be combined with an embryonic stem cell myocardial differentiation experiment for use. And the accuracy of predicating the developmental toxicity of the test substance is improved.

Provided are methods for in vitro induced differentiation of embryonic stem cells or pluripotent stem cells into dopaminergic neural progenitor cells, dopaminergic neural precursor cells and / or dopaminergic neurons. Also provided are media for use in the method, cell populations obtained using the method, and the use of the obtained cell populations in the prevention and / or treatment of diseases and / or dysfunctions associated with dopaminergic neuron degenerative disorders.

The application discloses a construction method of a spontaneous psoriasis mouse model and application thereof, and inserts a loxP site into an embryonic stem cell clone in a fatty acid synthase gene Fasn Chimeric mice are obtained through blastocyst injection, and through mating, a Fasn flox / flox Mouse; and a tool mouse Krt14-CreERT2 + / ‑ The tool mouse is mated with the tool mouse, and a mouse with a genotype of Krt14-CreERT2 + / ‑ ; Fasn flox / flox is screened, and the mouse model is obtained through continuous induction of tamoxifen. The spontaneous psoriasis mouse model constructed by the application is superior to an acute model which needs external induction, is convenient for long-term observation and intervention experiments, and provides a more ideal tool for researching a pathogenesis of psoriasis, screening and evaluating new drug candidate compounds for psoriasis.

The present application provides a method for inducing differentiation of embryonic stem cells into megakaryocytes. The provided method comprises the step of inducing differentiation of engineered embryonic stem cells into megakaryocytes; the engineered embryonic stem cells express a reduced amount of SHMT2 protein compared to unengineered embryonic stem cells. Megakaryocytes and platelets can be efficiently prepared via the method.

The application relates to a method and system for establishing a prediction model of chemical liver toxicity, which comprises the following steps: (1) differentiating human embryonic stem cells or human induced pluripotent stem cells into hepatocyte-like cells and constructing a three-dimensional hepatocyte model; (2) taking miR-122, LDH and Cyto C in the three-dimensional hepatocyte as combined test indexes to judge liver toxicity of a liver toxicity mode compound; (3) classifying the liver toxicity mode compound into three categories through two canonical discriminant functions by a linear discriminant analysis modeling method; and (4) converting the canonical discriminant function into a Fishers discriminant function to obtain a liver toxicity prediction model. The application has the general characteristics of a conventional cell model for evaluating compounds, and can also evaluate the toxicity effect and health risk of the compounds in a high-throughput manner with relatively less manpower.

Described herein are methods for generating human forebrain endothelial cells, compositions comprising the cells, and methods of use thereof in therapy. Provided herein are methods for generation of human embryonic forebrain-like endothelial cells (e.g., periventricular endothelial cells) from human embryonic stem cells; the methods include addition or GABA and WNT7A for efficient differentiation, and isolation of GABRB37CD31+ cell population by FACS.

The invention discloses a large-scale stem cell culture method, and relates to the technical field of stem cell culture, and the method comprises the following steps: stem cell pretreatment: taking P3-P5 generation mesenchymal or embryo dry and thin parts, digesting for 2-3 minutes at 37 + / -0.5 DEG C by using trypsin containing 0.01%-0.05% of EDTA (Ethylene Diamine Tetraacetic Acid), the pretreated cells are inoculated to a bioreactor containing a modified microcarrier, the microcarrier is a chitosan and gelatin composite material, the aperture is 50-100 microns, the particle size is 200-300 microns, RGD peptide is grafted, dynamic fed-batch culture is conducted, the concentration of glucose and lactic acid is detected in real time through an intelligent feedback system, harvesting and purification are conducted, 0.02%-0.04% of collagenase is added after culture is finished, incubation is conducted for 15-20 min at the temperature of 37 DEG C, and the concentration of glucose and lactic acid in the bioreactor is detected. The limitation of a traditional single-material microcarrier is broken through, the chitosan and gelatin composite carrier is adopted, RGD peptide is grafted, the cell attachment rate is increased, a three-dimensional growth space is provided for cells through specific aperture / particle size design, and the problem of poor cell uniformity in large-scale culture is solved.

The invention provides a skin organoid HPV infection model based on a human embryonic stem cell source and a construction method and application thereof, and relates to the technical field of molecular biology, the construction method comprises the following steps: integrating an HPV genome to an AAVS1 site of genome DNA of a human embryonic stem cell to obtain the human embryonic stem cell carrying the HPV genome, and performing induced differentiation and activation of the HPV genome to obtain the human embryonic stem cell carrying the HPV genome. A skin-like organ HPV infection model is formed, stable heredity and controllable expression of an HPV genome are achieved, virus particle assembly is achieved through induced differentiation, establishment of a virus replication environment, activation of HPV early gene expression, driving of HPV genome replication and induction of late gene expression, and the natural infection state of HPV is simulated. The technical problem that in the prior art, an infection model cannot simulate the life cycle of HPV dependent terminal differentiation is solved, and an irreplaceable technical platform is provided for research of an HPV pathogenic mechanism and research and development of antiviral drugs and vaccines.

The present application relates to a kind of enrichment method of stem cell high purity positive cell group, belong to the field of biotechnology and cell engineering technology.It aims to solve the technical problems such as the cell damage of existing screening technology, low screening efficiency, easy to affect stem cell pluripotency and genome stability, etc.The present application forms 23-cell small mass after H9 human embryonic stem cells infected by lentivirus are digested gently, after gradient dilution static culture, high expression positive clone is screened under fluorescence microscope, and target cell group is directly enriched by visual mechanical scraping.The present application does not change cell genome, and is high in safety;Small mass inoculation significantly improves cell survival and cloning efficiency at low density;Selective scraping operation is accurate, fast and low in cost;The proportion of reporter gene (EYFP) positive cells in the obtained positive cell group can reach 93.2%, and good pluripotency and function are maintained, suitable for efficient enrichment of stem cells and immune cells.

This invention provides a method for culturing embryonic stem cells from Orobanchaceae plants. The steps include: obtaining explants from Orobanchaceae plants; inducing culture of the explants to obtain embryonic stem cells; and inoculating the embryonic stem cells into an expansion medium containing Artemisia annua extract for further culture. This invention provides, for the first time, a method for the isolation and rapid culture of embryonic stem cells from Orobanchaceae plants. This method is independent of host plants and can rapidly and massively obtain Orobanchaceae plant stem cells under artificial conditions, fundamentally solving the problem of ecological damage caused by traditional production methods. For the first time, Artemisia annua extract is applied to the culture of Orobanchaceae plant stem cells. Utilizing its natural antibacterial and bioactivity, it not only effectively inhibits microbial contamination during the culture process but also unexpectedly shows that it can significantly promote the growth of Orobanchaceae plant stem cells, shorten the culture cycle, and reduce production costs.

Methods are provided for detection of a target cell type within a cell population, and compositions are provided comprising cells and an indicator that indicates the number of cells of the target cell type in the cell population. Examples are provided in which these methods are used to detect human embryonic stem cells within a differentiated cell population with exquisite sensitivity. Differentiated cells produced from embryonic stem cells can be characterized by these methods before transplantation into a recipient, thereby providing further assurance of safety.

An amplifying method of pancreatic cells is provided. The amplifying method includes performing digestion, resuspension, discontinuous density gradient centrifugation treatment and amplifying treatment sequentially. The mammalian pancreatic duct is used as the source of pancreatic precursor-like cells in the amplifying method, and islet cells and acinar cells in the cell clusters obtained by the discontinuous density gradient centrifugation treatment are removed. It is beneficial to improve the yield of the pancreatic precursor-like cells availably, and avoid the ethical restrictions and possible carcinogenic risks caused by using the embryonic stem cells. The amplifying medium used comprises a reprogramming substance composed of several small molecule compounds. It can avoid the risks of non-specific and off-target deletion that are easily caused by the use of the gene-editing methods to change the gene sequence. Also provided is a differentiation method and an application of the pancreatic precursor-like cells obtained by the amplifying method.