204 results about "Embryonic stem cell" patented technology

The invention discloses a pig chimeric embryo culture medium and application thereof in effectively improving the in-vitro chimeric ability of pig embryonic stem cells, and belongs to the technical field of bioengineering. According to the pig chimeric embryo culture medium disclosed by the invention, a neutral culture environment for promoting synchronous development of stem cells and embryos is created by accurately regulating and controlling growth factors, nutrient substances and other key components in the culture medium. On the basis, overexpression of anti-apoptosis factors, optimization of microinjection conditions and adoption of a laser-assisted incubation means are combined, so that the chimeric efficiency is further improved, integration of the stem cells and embryos is effectively supported, the chimeric efficiency is improved, and a reliable in-vitro tool is provided for evaluating the chimeric ability and pluripotency of the pig stem cells.

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 stem cell biology, and relates to pedigree specific differentiation of human pluripotent or embryonic stem cells, in particular to a mesenchymal stem cell induced differentiation culture medium and application thereof. The mesenchymal stem cell induced differentiation culture medium is used for directly inducing and differentiating human induced pluripotent stem cells / or embryonic stem cells into mesenchymal stem cells, the culture medium comprises a mesenchymal stem cell basic culture medium and an additive, and the additive comprises the following components: 2-10 [mu] M of a TGF-beta inhibitor, 2-10 [mu] M of a GSK3 beta inhibitor and 5-10 [mu] M of a Rock inhibitor. By adopting the induced differentiation culture medium provided by the invention, human induced pluripotent stem cells / or embryonic stem cells can be directly induced and differentiated into mesenchymal stem cells, and the obtained mesenchymal stem cells have the advantages of infinite sources, stable quality, high homogeneity and the like; and the human induced pluripotent stem cells / or embryonic stem cells can be directly and directionally differentiated into the mesenchymal stem cells in a relatively short time, so that the culture time is shortened, and the production cost is reduced.

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.

Provided are a method for producing a porcine blastoid from porcine embryonic stem cells (pESC), and a blastoid obtained by the method. Also provided are media for inducing the pESC to form blastula-like embryos and media for producing and culturing the pESC.

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.

Disclosed are compositions, in particular, organoid compositions, derived from more than one donor cell. Further disclosed are methods of making compositions, for example, organoid compositions, that comprise a differentiated cell population derived from more than one donor cell. Donor cells may include, for example, a precursor cell such as an embryonic stem cell or other precursor cell. The disclosed methods use synchronization conditions to produce a synchronized pooled-precursor cell population, which may then be differentiated into an organoid composition. Methods of using the compositions are also disclosed.

The invention relates to a preparation method of human haploid trophoblast stem cells, which comprises the following steps: inoculating haploid embryonic stem cells (hAG-ESCs) into a culture medium of human trophoblast stem cells (TSC), domesticating, and when TSC sample clones account for more than 50% of the proportion of all cells in the induction process, carrying out flow sorting on ITGA6 positive cells, and inoculating into the TSC culture medium again for culturing to obtain the human haploid trophoblast stem cells. After the culture, haploid trophoblast stem cells (haiTSCs) are obtained; the prepared haiTSCs are haploids, and have TSC properties. The preparation method is rapid and efficient, and the haiTSCs can be used for simulating an in-vivo disease model and have the genetic screening advantage of haploids.

The invention relates to the technical field of cell biology, and particularly discloses a goat embryonic stem cell culture system, a system establishment method and application, the goat embryonic stem cell culture system takes mTeSRTM Plus as a basic culture solution, and 20 ng / mL to 25 ng / mL of leukemia inhibition factors, 1 mu M to 1.5 mu M of PD0325901, 4 mu M to 5 mu M of IWR-1, 45 mu g / mL to 50 mu g / mL of Vc, 15 ng / mL to 20 ng / mL of activin A, 1 mu M to 1.2 mu M of CGP77675 and 1.5 mu M to 2 mu M of GO6983 are added. Cell clone with clear edges is obtained through culture of a goat embryonic stem cell culture system, stable passage can be achieved, and the cell clone has the potential of being differentiated into three germ layers.

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.

To provide stem cell therapies for treating (curing) a variety of diseases or conditions, as an alternative to overcome certain shortcomings of existing embryonic stem cells and iPS cells.SOLUTION: Provided herein are: precursory regulatory cytotrophoblast cells produced in vitro and expressing specific cell surface proteins and combinations thereof; compositions thereof; and methods of treating disorders (e.g., cancers, inflammations, or autoimmune diseases) or conditions (e.g., skin conditions) by using the cells and compositions.SELECTED DRAWING: None

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 present invention provides populations of photoreceptor rescue cells (PRCs) having a unique profile of markers and resulting from in vitro differentiation of early progenitor cells, including pluripotent stem cells and embryonic stem cells (ESCs). Also provided are methods of generating populations of photoreceptor-rescuing cells and methods of using them to treat ocular conditions.

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.

Disclosed herein is a genetically modified rodent, the genetically modified rodent comprising a humanized Clec9a gene in its genome. The invention also discloses an isolated rodent tissue or cell. The isolated rodent tissue or cell comprises a rodent embryonic stem cell and a rodent embryo; a method of making the genetically modified rodent; and methods of using the genetically modified rodents.

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.

A method for preparing hematopoietic stem cells and progenitor cells is provided for producing hematopoietic stem / progenitor cells (HSPCs) from embryonic stem cells or iPSCs. Also provided are a hematopoietic stem / progenitor cell (HSPC) culture product generated by the method, and the use of differentiating the hematopoietic stem / progenitor cells (HSPCs) into blood cells of interest. The method successfully achieves efficient differentiation into HSPCs, and the obtained culture product does not contain heterologous substances, meets the GMP standard, and is prone to large-scale production. Additionally, the method also has the advantages of shorter differentiation time and higher efficiency.

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 discloses a culture medium and a culture method of araliaceae plant embryogenic stem cells and a method for producing saponin at high yield by using the araliaceae plant embryogenic stem cells. The culture medium comprises a basic culture medium, and 200 mg / L-500 mg / L of cerium nitrate is added into the basic culture medium; the basic culture medium is selected from one of an MS culture medium and a B5 culture medium. The MS culture medium is further added with amphotericin B with the concentration of 0.8 to 1.3 mg / L and double antibodies with the mass concentration of 0.9 to 1.2 percent, and the double antibodies are 100X mycillin mixed liquor, 6-BA with the concentration of 0.8 to 1.2 mg / L and NAA with the concentration of 0.8 to 1.3 mg / L; or 0.098 mg / L to 0.120 mg / L of 6-BA, 13 mg / L to 18 mg / L of cane sugar, 0.48 mg / L to 0.52 mg / L of methyl jasmonate and 0.099 mg / L to 0.130 mg / L of triacontanol are also added into the B5 culture medium. The cerous nitrate is added into the culture medium of the araliaceae plant embryonic stem cells, the culture of the araliaceae plant embryonic stem cells is obviously promoted, and the culture quantity of the embryonic stem cells can be obviously increased, so that efficient and rapid culture is realized, and the culture medium has universality.