156 results about "Embryonic stem cell" patented technology

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 belongs to the field of antibody development, and discloses a screening method and application of a human-mouse protein high homologous target antibody based on a fully humanized antibody mouse. Aiming at the problem of weak antibody response caused by immune tolerance of human-mouse high homologous targets (protein homology is greater than or equal to 95%), the following scheme is provided: in embryonic stem cells (ES cells) of HUGO-Mabfully humanized antibody transgenic mice, a mouse target gene (such as ACVR2A) is knocked out through a Turbo Knockout technology, and homozygous knockout ES clones are screened; carrying out microinjection on the clones to the whitened B6 mouse blastocyst, and transplanting a pregnant mouse to obtain a Founder mouse; the Founder mouse is subjected to target antigen immunization for more than or equal to 4 times (the Freund's complete adjuvant is used for the first time), and the titer of the serum antibody is detected. According to the invention, 100% homozygous knockout chimeric efficiency is realized in the Founder stage, the mouse construction period is shortened from traditional 8-10 months to 3-4 months, and the diversity and affinity of the antibody are significantly improved (titer reaches 1: 729,000). The obtained antibody can be used for preparing medicines for treating tumors or autoimmune diseases.

Disclosed in the present invention is a use of alisol-B23-acetate (AB23a) in the preparation of a drug for preventing or treating hypertrophic cardiomyopathy and diseases caused by the hypertrophic cardiomyopathy. The present invention provides for the first time a use of AB23a in the prevention or treatment of hypertrophic cardiomyopathy. According to the present invention, use of AB23a in direct in vitro treatment of cardiomyocytes derived from directed differentiation of human embryonic stem cells revealed that AB23a-based in-vitro treatment can significantly inhibit hypertrophic phenotypes of the human embryonic stem cells-cardiomyocytes; using an AB23a-containing feed to feed mice with hereditary hypertrophic cardiomyopathy caused by a gene mutation revealed that AB23a prominently relieves pathological myocardial hypertrophy of the mice and improves the cardiac function; AB23a can be used for preparing a drug for resisting hereditary hypertrophic cardiomyopathy, offering a novel way and means for treating hypertrophic cardiomyopathy; and AB23a is the main medicinal ingredient in the traditional Chinese medicinal herb-Rhizoma Alismatis, is safe to organisms, and has good clinical application prospects.

The invention belongs to the technical field of medicines, and discloses application of exosomes derived from human pluripotent stem cells in preparation of anti-hepatic inflammation medicines. According to the invention, the exosome derived from the human pluripotent stem cells is applied to the preparation of the anti-hepatic inflammation medicine, especially the preparation of the anti-hepatic fibrosis anti-inflammatory medicine. The human embryonic stem cell-derived exosome can inhibit and regulate polarization of liver macrophages, inhibit inflammatory liver macrophages, up-regulate anti-inflammatory liver macrophages, reduce secretion of inflammatory factors, improve secretion of anti-inflammatory factors and inhibit liver inflammations and development thereof, has an obvious anti-inflammatory treatment effect on inflammation of liver fibrosis, and can be used for preparing medicines for treating liver fibrosis. Therefore, the progress of hepatic fibrosis can be inhibited.

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The present invention discloses an application of highly active human umbilical cord mesenchymal stem cells. HA-MSCs were isolated from human umbilical cord tissue using TeSR-E8 containing 5% CloneR and LN521-Coated culture flasks, and a technical system for HA-MSC isolation, expansion, and cryopreservation was established. Compared with conventionally cultured MSCs, the HA-MSCs prepared by the present invention have higher proliferation activity, smaller size, a higher nuclear-cytoplasmic ratio, and highly express embryonic stem cell-associated marker antigens SOX2, Nanog, and OCT4. They have the potential to differentiate into neural, myocardial, and hepato-intestinal progenitor cells derived from the three germ layers and possess excellent anti-aging activity.

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.

This disclosure provides an improved system for culturing human embryonic stem cells. The cells are cultured in suspension so as to maximize the production capacity of the culture environment. The new culture system of this invention allows for bulk proliferation of hES cells in a more cost-effective manner, which facilitates commercial production of important products for use in human therapy.

The invention discloses a culture medium composition for inducing human pluripotent stem cells to differentiate into human iAGM hematopoietic organoid, human embryonic stem cells are differentiated into the human iAGM hematopoietic organoid through combination and matching of different cell factors in stages, and the generated hematopoietic stem cells have high primitiveness and authenticity, so that the functionality of the hematopoietic stem cells is finally guaranteed.

Disclosed herein are methods, compositions, and culture media for generating synthetic embryos in vitro from mammalian pluripotent stem cells, such as pluripotent embryonic stem cells. In some embodiments, the methods can include co-culturing wild-type mammalian pluripotent stem cells with modified mammalian pluripotent stem cells that include one or more genes encoding transcription factors that can drive the generation of extraembryonic or extraembryonic-like cells (e.g., GATA6, SOX17, GATA3, and / or TFAP2C genes) in a culture medium under conditions that allow the pluripotent stem cells to self-organize into post-implantation embryonic structures. In some embodiments, the pluripotent embryonic stem cells are human pluripotent embryonic stem cells, and the generated synthetic embryo is a human embryo.

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 provides a spot universal CFR64-T cell prepared on the basis of CRISPR / Cas9 as well as a preparation method and application thereof. The CFR64-T cell is obtained by induced differentiation of recombinant human embryonic stem cells; in the recombinant human embryonic stem cell, a coding sequence of a CFR64 molecule is knocked into a TRAC site of the human embryonic stem cell at a fixed point through a CRISPR / Cas9 editing system; the CFR64 molecule comprises an antigen binding structural domain, a transmembrane structural domain and an intracellular costimulatory signal structural domain, the antigen binding domain is a human Fc gamma receptor extracellular domain, and the amino acid sequence of the antigen binding domain is shown in SEQ ID NO: 1. The CRISPR / Cas9 technology is adopted, gene integration sites are controllable and free of oncogene mutation risks, TCR genes are knocked out, the immunological rejection risk of universal CFR64-T cells to hosts is reduced, and meanwhile gene expression is more stable.

The invention relates to a thermostable FGF10 polypeptide possessing FGF10 activity and having or comprising at least 85% sequence identity to SEQ ID NO:3 that has or comprises an amino acid sequence from Ser69 to Ser208 of SEQ ID NO:1, or to SEQ ID NO:5 that has or comprises an amino acid sequence from Leu40 to Ser208 of SEQ ID NO:1, or the fragments thereof, comprising at least an amino acid substitution L152F. Preferably, any of amino acid substitutions V123I, Q175E, and N181D can be further included. The invention further discloses the use of subjected thermostable FGF10 polypeptides in regenerative medicine or other related medical applications or cosmetics. Further, it discloses a culture medium comprising subjected thermostable FGF10 polypeptides suitable for proliferation and differentiation of the human embryonic stem cells or formation and differentiation of spheroids and organoids.

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 belongs to the technical field of circular RNA, and discloses application of circular RNA derived from human pluripotent stem cell exosome in preparation of anti-inflammatory drugs for hepatic fibrosis. The invention relates to an application of human pluripotent stem cell exosome-derived circular RNA (Ribonucleic Acid) in preparation of an anti-inflammatory drug for hepatic fibrosis, in particular to CircBase ID of the circular RNA: hsacirc0076798. The human pluripotent stem cell exosome-derived circular RNA is circRNA in a human embryonic stem cell exosome, has the effects of regulating macrophage phenotype and inhibiting inflammatory response, and provides a thought and a method for preparing medicines for preventing and treating inflammation and reversing liver fibrosis. The circular RNA is used as a molecule for regulating and controlling gene expression of liver macrophages, and is expected to be used for preparing anti-inflammatory targeted drugs for hepatic fibrosis.

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 relates to the technical field of biology, and discloses a culture medium system for pluripotency maintenance and directional induction of embryonic stem cells and application of the culture medium system. 0.5 to 0.8 part of tetramethylenediamine; 0.2 to 0.5 part of corpus luteum hormone; 1-2 parts of a vitamin mixture; 2-4 parts of bovine serum albumin; 0.5 to 1.0 part of sodium bicarbonate; 0.2 to 0.4 part of an antibiotic mixed solution; 1 to 2 parts of biotin; 0.5 to 1.0 part of bone morphogenetic protein; and 0.1 to 0.2 part of magnesium ethylene diamine tetraacetate. A multi-factor regulation and control system is constructed, signal inhibition, a three-dimensional scaffold and a metabolic regulation module are combined, pluripotency of the embryonic stem cells is stably maintained, culture uniformity, structural support and differentiation resistance are remarkably improved, and the technical bottleneck of a traditional system in the aspects of induction efficiency and stability is broken through.

The invention provides a method for preparing an antitumor drug by inducing differentiation of neuroblastoma through co-culture supernate of human embryonic stem cells and neuroblastoma cells. The human embryonic stem cells have the biological characteristics of multidirectional differentiation potential, infinite proliferation, self-renewal and the like, and the microenvironment of the human embryonic stem cells can reprogram cancer cells. The early neuroblastoma can naturally fade, and research shows that cancer cells of the neuroblastoma retain certain differentiation potential and can be differentiated towards neurons. The inventor discovers that the neuroblastoma cells and embryonic stem cells are subjected to direct contact co-culture, and the generated co-culture supernatant can induce the neuroblastoma cells to differentiate into neuronal cells, so that the proliferation, migration and invasion of the neuroblastoma cells are inhibited in vitro. The discovery shows that the co-culture supernate has huge potential in innovative biological treatment application of resisting neuroblastoma.

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 provides a differential medium composition and a method for preparing mesenchymal stem cells by inducing stem cells to differentiate in a body segment direction, and belongs to the field of biology. The differential culture medium combination comprises a culture medium A, a culture medium B, a culture medium C and a culture medium D. The culture medium A comprises a basic culture medium and a first small molecule compound combination, the culture medium B comprises a basic culture medium and a second small molecule compound combination, and the culture medium C comprises a basic culture medium and a third small molecule compound combination. The culture medium D comprises a combination of the basic culture medium and a fourth small molecule compound. According to the differential medium composition and the induction method provided by the invention, the mesenchymal stem cells can be prepared from the pluripotent stem cells or the embryonic stem cells.

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.

To provide genetically engineered stem cells, plasma cells, and B cells for avoiding immune rejection within a host, and methods of making the same and uses thereof.SOLUTION: Provided is a method of differentiating human embryonic stem cells (ES cells) into plasma cells, the method comprising producing precursor cells and differentiating the precursor cells into B cells.SELECTED DRAWING: Figure 10

The invention of a reactor for the production of growth factors to regenerate the epithelium of the auditory system from embryonic stem cells, aimed at stimulating sensory tissue to enhance hearing capacity, in the form of a lyophilized powder provides a novel reactor and method for producing growth factors that stimulate stem cells in the auditory region, promoting their conversion into auditory epithelial cells. The reactor comprises three sections: an upper transparent section for culture observation and sterilization via a UV lamp, a middle section for continuous mixing and purification using micro-filters, and a lower section for lyophilization. The reactor maintains optimal conditions for stem cell proliferation, utilizing controlled temperature and vacuum pressure to concentrate beneficial substances and crystallize growth factors into a lyophilized powder. This product serves as a potential treatment for auditory conditions, enhancing the regeneration of auditory structures and delaying the onset of presbycusis.

The invention relates to a method for improving myocardial protection activity of human embryonic stem cell exosomes and application of the method, and belongs to the technical field of biological medicine. In order to solve the problem of low biological activity of embryonic stem cell exosomes extracted through in-vitro culture, the invention provides a method for improving myocardial protection activity of human embryonic stem cell exosomes, when human embryonic stem cells are subcultured until the confluence degree reaches 70-80%, green light is used as a unique light source to vertically irradiate the human embryonic stem cells for a certain period of time, and the human embryonic stem cell exosomes are obtained. Continuing to culture the human embryonic stem cells subjected to light stimulation, and centrifugally extracting the exosomes generated by the human embryonic stem cells. According to the method, the yield of the human embryonic stem cell exosome is increased, the myocardial protection activity of the human embryonic stem cell exosome is remarkably improved, and the exosome generated by the human embryonic stem cell irradiated by green light can remarkably promote myocardial cell proliferation, inhibit myocardial cell apoptosis caused by hypoxia, reduce the myocardial infarction area and recover the infarction heart function.

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.

The present invention discloses a method for guiding pluripotent stem cells to become developmentally arrested cells and its application, relating to the technical field of stem cells. The method for guiding pluripotent stem cells to become developmentally arrested cells includes: continuously culturing in a medium for culturing pluripotent stem cells with rapamycin added thereto. By adding rapamycin in the culture of pluripotent stem cells (such as human embryonic stem cells), it is possible to guide pluripotent stem cells to become cells that are developmentally arrested in a natural state, which is beneficial to preventing the excessive proliferation of pluripotent stem cells in special circumstances, provides new ideas for more applications of human pluripotent stem cells, and can improve the efficiency of cell production; it is beneficial to obtain pluripotent stem cells with a specific degree of development as needed, providing more possibilities for the application of human pluripotent stem cells.

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.