162 results about "Pluripotential stem cell" patented technology

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

The invention provides a pancreatic islet organ capable of efficiently maintaining pancreatic islet beta cell identity and a preparation method and application thereof. Specifically, the invention provides the preparation method for the in-vitro pancreatic islet organ, and the preparation method comprises the following steps: carrying out induced differentiation culture on the ZnT8-knocked-out pluripotent stem cells, so as to obtain the mature pancreatic islet organ. The invention also provides a corresponding transplantation composition and application thereof. The invention discloses that ZnT8 knockout is a key factor for effectively maintaining the identity stability of beta cells in pancreatic islet organs. In addition, the pancreas islet beta cell identity maintenance agent is combined with the ZnT8-knocked-out pancreas islet organ for use, so that the identity stability of the pancreas islet organ beta cell can be obviously maintained. The invention provides a new strategy for transplantation treatment of type 2 diabetes, and has excellent clinical transformation value.

The application discloses a construction method and application of a thymus organoid, and the method is used in combination with small molecule compounds and small molecule proteins to differentiate pluripotent stem cells into the thymus organoid under a suspension culture condition, and the method can efficiently obtain the thymus organoid which is closer to the natural thymus in structure and function.

The invention discloses a method for generating haploid sperm-like cells through in-vitro differentiation of pluripotent stem cells and application of the haploid sperm-like cells. Human pluripotent stem cells are firstly differentiated into ectoderm-like cells, then the ectoderm-like cells are spontaneously gathered and converted into primordial germ cell-like cells under the induction of ectoderm induction factors, and then the primordial germ cell-like cells and testicular sertoli cells are subjected to mixed culture to form recombinant testicular tissues; the recombinant testis tissue enters meiosis under the induction of nutrition limitation and retinoic acid, then meiosis is completed under the stimulation of meiosis second-stage induction factors such as testosterone, follicle-stimulating hormone and pituitary extract, and interaction between primordial germ cell-like cells and testis supporting cells is enhanced by using small molecules SB431542, so that the meiosis of the testis is enhanced. And finally inducing to generate haploid sperm-like cells. According to the invention, a new tool can be provided for researching a molecular mechanism of male reproduction, and an important thought can be provided for research and transformation of asthenospermia, oligospermia, drug screening and the like.

Provided is an isolated human naive pluripotent stem cell (PSC), wherein: (i) when the naive PSC is a female PSC, then said naive female PSC has two unmethylated alleles of an X-inactive specific transcript (XIST) gene; and (ii) when said naive PSC is a male PSC, then said naive male PSC has an unmethylated allele of said XIST gene. Also provided is a culture medium which comprises an ERK1 / 2 inhibitor, a GSK3beta inhibitor, a p38 inhibitor, a JNK inhibitor, a STAT3 activator and at least one agent selected from the group consisting of: bFGF, TGFbeta 1, a PKC inhibitor, a ROCK inhibitor and a NOTCH inhibitor; or at least agent selected from the group consisting of: a TGFR inhibitor, a FGFR inhibitor, a PKC inhibitor, a ROCK inhibitor and a NOTCH inhibitor.

The present invention relates to CD73 + CD44 + , CD90 + A method for generating a population of mesenchymal stromal cells (MSCs) is provided. + CD44 + , CD90 + MSCs are used in methods to generate terminally differentiated osteogenic, adipogenic, and chondrogenic cells from pluripotent stem cells (PSCs), which involve the use of a single agent, a WNT signaling pathway activator (e.g., a GSK3β inhibitor), in adherent cultures of PSCs.

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.

The present disclosure relates generally to methods and systems of producing Schwann cells from pluripotent stem cells under fully defined conditions. The Schwann cells produced by the disclosed methods find applications as models of the enteric nervous system, tools for high throughput screening of potential therapeutics for treatment of enteric neuropathies, and in regenerative medicine.

To provide a method for evaluating the quality of neuroretina for transplantation and neuroretina sheets for transplantation selected by said method. [Solution] The present invention provides a method for evaluating the quality of neuroretina for transplantation, comprising: extracting a part or all of a cell aggregate containing a neuroretina having an epithelial structure derived from pluripotent stem cells as a quality evaluation sample; detecting the expression of neuroretinal cell-related genes and non-neuroretinal cell-related genes in the quality evaluation sample; and determining, if the expression of neuroretinal cell-related genes is observed and the expression of non-neuroretinal cell-related genes is not observed, that (1) a neuroretina (neuroretina for transplantation) from the same cell aggregate as the cell aggregate containing a part of the quality evaluation sample, (2) a neuroretina (neuroretina for transplantation) from the same lot as the cell aggregate containing a part of the quality evaluation sample, or (3) a neuroretina (neuroretina for transplantation) from the same lot as the cell aggregate of the entire quality evaluation sample, wherein the non-neuroretinal cell-related genes include one or more genes selected from the group consisting of brain and spinal cord tissue marker genes and eyeball-related tissue marker genes.

A novel method of inducing or producing hepatocytes from human induced pluripotent stem cells at an unprecedented efficiency and functionality. The core of the invention is the use of experimentally discovered mRNAs at multiple critical differentiation decision points along a pluripotent to mesendoderm to endoderm to hepatocytes pathway in a previously unknown manner.

The application provides a freezing and recovery method and application of pluripotent stem cell induced differentiation middle stage cells, and the freezing method comprises the following steps: (1) the starting cells are pluripotent stem cells, which are induced to the middle stage; (2) the cells are recovered and counted by using a mild digestion enzyme, and the cells are resuspended in a freezing solution; (3) after programmed cooling to-80 DEG C, the cells are transferred into liquid nitrogen for storage, and are ready for use. The freezing method and the freezing solution provided by the application significantly improve the freezing survival rate of the differentiation middle stage cells, avoid the apoptosis or irreversible stress damage induced by the traditional freezing solution, and obviously improve the problems of cell damage and low survival rate caused by the general freezing solution in the prior art. The differentiation potential of the recovered cells is good, and the cells can successfully form mature kidney unit structures, so that the problem of subsequent differentiation failure in the prior art is solved.

Disclosed herein are enteroids, obtained by dissociating an epithelial cell population from a non-transplanted human intestinal organoid (HIO), such as an HIO produced in vitro from human pluripotent stem cells (hPSCs). Also disclosed herein are methods of producing and using the same, including methods of transplantation involving said enteroids, particularly for treating a gastrointestinal disease or condition. Also disclosed herein are methods of transplantation involving HIOs and / or HIO components, particularly for treating a gastrointestinal disease or condition.

The invention provides application of SP140 in regulating and controlling differentiation of pluripotent stem cells to hematopoietic stem cells and / or hematopoietic progenitor cells, and relates to the technical field of cell drug preparation. The expression of SP140 in human pluripotent stem cells (hPSC) or in the differentiation process of the human pluripotent stem cells is reduced on the gene level and the protein level, and the human pluripotent stem cells are promoted to be differentiated into human hematopoietic stem cells (HSC) with multiple transplantation potential. The method not only realizes in-vitro regeneration of the hematopoietic stem cells and / or hematopoietic progenitor cells, but also has the characteristics of short differentiation period, high differentiation efficiency and reduction of culture time and cost, and the prepared hematopoietic stem cells express typical biomarkers and have good multi-transplantation chimeric ability. The invention provides a new way for the source of hematopoietic stem cells required by regenerative medicine and treatment of blood and related diseases.

The present invention provides a pluripotent stem cell into which a nucleic acid encoding a chimeric antigen receptor (CAR) specific to CD5, CD2, or CD6 has been introduced. The present invention also provides a CAR-T cell specific to CD5, CD2, or CD6, which is obtained by inducing the differentiation of the pluripotent stem cell into a T cell, and has the following characteristics: (a) that the expression of CD5, CD2, and CD6 is reduced compared to that of a corresponding CAR-T cell derived from peripheral blood; and (b) that cytotoxic activity against T-cell tumors is higher than that of the corresponding CAR-T cell and a CD5, CD2, or CD6 gene-deficient CAR-T cell, which are derived from peripheral blood.

The invention belongs to the technical field of biological medicines, and particularly discloses application of mesenchymal stem cells derived from ipsc induced pluripotent stem cells in systemic lupus erythematosus. The ipsc is engineered by using a genetic engineering technology to prepare an ipsc cell with a stable over-expression CD19CAR structure, and the ipsc cell is induced by using an EB method to form the mesenchymal stem cell. The finally formed CD19 CAR mesenchymal stem cell can target B cells, and the CAR structure can be activated by an antigen to enhance the immune effect, and meanwhile, the effects of targeting and enhancing the immune effect are achieved, so that the purpose of treating systemic lupus erythematosus is achieved.

The present invention refers an in vitro method for inducing differentiation of human pluripotent stem cells (hPSCs) generating hindbrain neurons and compositions thereof. The invention also refers to the hindbrain neurons and compositions thereof for use in diagnostic screening and for use in the prevention and / or treatment of neurological disorders.

The invention provides a composition, cell culture and application of the composition in differentiation of human pluripotent stem cells to myocardial cells and / or construction of human-derived heart organoid, and belongs to the technical field of cell engineering. The invention provides a composition. The composition comprises a fatty acid receptor antagonist, a fatty acid synthetase inhibitor and a fatty acid oxidation activator. The composition provided by the invention can regulate cell metabolism, breaks through the contradiction that the proliferation capacity and the cell maturity cannot be obtained at the same time in the traditional technology, can maintain the myocardial cells in a proliferation stage, and obtains the myocardial cells with relatively mature structures and functions, and the contractility of 3D heart organs constructed by the myocardial cells is greatly improved. The invention provides a high-quality cell source for myocardial regeneration treatment, high-throughput drug cardiotoxicity screening and heart disease mechanism research, and has remarkable clinical application value and industrialization prospect.

This cell-based formulation contains SSEA-3-positive pluripotent stem cells derived from mesenchymal tissue of a living body or derived from cultured mesenchymal cells. This cell-based formulation is characterized in that the formulation is for administration to address diseases and / or post-acute sequelae caused by SARS-CoV-2 infection. The present invention makes it possible to provide a cell-based formulation that contains pluripotent stem cells and is used for treating and / or preventing SARS-CoV-2 infection-caused diseases such as pneumonia and pulmonary fibrosis and SARS-CoV-2 infection-caused post-acute sequelae such as olfactory dysfunctions.

Provided are: a method for producing a spinal cord organoid from a pluripotent stem cell in vitro, the method comprising (1) a step for culturing a human pluripotent stem cell in a culture medium that contains a ROCK inhibitor and a Wnt signal activator but does not substantially contain a TGFβ inhibitor or a BMP signal inhibitor, (2) a step for further culturing the cell obtained in step (1) in a culture medium that contains a retinoic acid receptor agonist and a hedgehog signal activator, and (3) a step for further culturing the cell obtained in step (2) in a culture medium that does not substantially contain a TGFβ inhibitor, a BMP signal inhibitor, a retinoic acid receptor agonist, or a sonic hedgehog stimulator to obtain a spinal cord organoid; and a spinal cord organoid obtained by the method. The spinal cord organoid according to the present application comprises at least a portion of the dorsal side of the spinal cord and a ventral region of the spinal cord.

A nanoparticle for delivery of mRNA to cell, comprising a core comprising a polyethylenimine polymer having fluorinated groups covalently coupled thereto and with mRNA reversibly associated therewith, and a shell surrounding the core comprising heparin. Ij some embodiments, the nanoparticle comprises a targeting agent associated with the shell, wherein the targeting agent is selected from the group consisting of agents that bind to receptors overexpressed on tumor cells, agents that bind to cell surf ace antigens that are expressed on pluripotent stem cells, agents that bind to cell surface antigens on T cells, and agents that bind to antigens presented by MHO molecules. Pharmaceutical compositions that include the nanoparticle and methods for using the nanoparticle for transfecting cells are provided

The present disclosure generally relates to methods and in vitro systems for generation of mitochondria-depleted pluripotent stem cells, composite pluripotent stem cells, or non-human, tetraploid, mitochondria-depleted embryos. The disclosure further provides methods for use of cells or non-human embryos in regenerative medicine and drug screening.

Provided are a production method of retinal pigment epithelial (RPE) cells that improves differentiation induction efficiency of pluripotent stem cells into RPE cells, and can provide highly pure RPE cells by a simple and easy operation in a short period, a culture method of RPE cells that can stably grow and culture a cell, a toxicity / efficacy evaluation method using RPE cells useful for transplantation therapy, and a therapeutic drug for a retinal disease. The invention relates to a production method of RPE cells, comprising adhesion culture of human pluripotent stem cells using a culture substrate coated with a laminin-E8 fragment, a culture method of RPE cells, comprising adhesion culture of RPE cells using a culture substrate coated with a laminin-E8 fragment, a toxicity or efficacy evaluation method using RPE cells obtained by producing or culturing by the method, and a therapeutic drug for a retinal disease, containing the RPE cells.

Provided are a chondrogenic stem cell for promoting joint repair and use thereof. Specifically, disclosed is a cell preparation used for alleviating or treating osteoarthritis, comprising a Procr-positive pluripotent stem cell. The Procr-positive chondrogenic stem cell can differentiate and generate downstream chondrocytes in vivo and promote cartilage repair in osteoarthritis damage, which will be a more effective cell treatment method. In addition, Procr proteins can be used as a new target of a drug for treating osteoarthritis.

The invention provides a construction method of an HBV (Hepatitis B Virus) infection model based on multilineage hepatobiliary organs, which comprises the following steps: differentiating human pluripotent stem cells to form immune cell-containing hepatobiliary organs for further maturation culture, and enabling the hepatobiliary organs to be mature in function and highly express NTCP (Natriuretic Therapeutic Cells) by using a maturation culture medium containing bile metabolism substrate cholesterol and deoxybear bile acid, so as to obtain the HBV infection model based on the multilineage hepatobiliary organs. And then infecting the mature organs by adopting an HBV virus strain generated by HepAD38 cells to construct an in-vitro model which is susceptible to HBV and can maintain an infection state for a long time. According to the construction method, the liver infection and immune interaction microenvironment can be simulated more truly, and a stable and reliable experimental platform is provided for HBV infection mechanism research and anti-HBV drug screening.

This invention discloses a method for inducing pluripotent stem cells to differentiate into skin keratinocytes and its culture medium, solving problems such as long differentiation cycles, low yields, insufficient purity, and animal-derived contamination in existing technologies. The first stage uses medium I containing FGF2 and tanshinone IIA to promote mesodermal differentiation. The second stage uses medium II containing BMP4, EGF, dexamethasone, and PVA to regulate and synergistically induce the BMP / ERK signaling pathway. The final stage uses medium III for expansion culture, obtaining fibroblasts with a purity >99% within 20 days. This method is non-invasive, simple, and low-cost, with potential for large-scale production. The obtained cells can be used for skin regeneration, disease modeling, drug screening, and clinical treatment, demonstrating significant medical application and industrial transformation value.

Provided herein are methods of generating hindbrain cells, including respiratory hindbrain cells, from pluripotent stem cells. Also provided are methods of generating a three-dimensional organoid comprising a population of hindbrain cells including a heterogeneous population of interneurons.

The present invention provides, as a technique that can be used for obtaining a skin organoid or a skin tissue structure that skillfully mimics the structure and function of the skin of a living body from stem cells, a skin organoid comprising cells that are derived from pluripotent stem cells and have the ability to differentiate into cells constituting the skin, the skin organoid having a lumen, the skin organoid having an epidermal cell layer comprising cells having the ability to differentiate into epidermis on the surface facing the lumen, and the skin organoid having a mesenchymal cell layer, on the outside of the epidermal cell layer, comprising cells having the ability to differentiate into dermis or subcutaneous tissue. The epidermal cell layer is configured by laminating, in this order from the lumen side, a granular layer comprising cells that express loricrin, a spinous layer comprising cells that express KRT10, and a basal layer comprising cells that express KRT5 and p63, and has hair pegs invaginating the mesenchymal cell layer.

The present invention relates to the use of corneal cell-derived mitochondria for the prevention or treatment of corneal damage or corneal endothelial cell diseases. It has been confirmed that mitochondria isolated from corneal endothelial cells differentiated from induced pluripotent stem cells (iPSCs) of the present invention alleviate the inflammatory response of corneal endothelial cells in an inflammatory environment, restore the integrity and function of corneal endothelial cells, and, when delivered into heterologous primary corneal endothelial cells, suppress inflammatory responses, particularly inflammation caused by physical damage, and promote the regeneration of corneal endothelial cells. Therefore, mitochondria possessing such effects can overcome the tumorigenic limitations of conventional stem cell therapies, and due to their ease of production and minimal regulatory requirements, they offer excellent cost-effectiveness and broad applicability.

The present invention provides a composition having a metabolism-improving action, which comprises a supernatant of brown adipocytes or a purified product thereof. The present invention also provides a method of preparing the supernatant without using a culture solution comprising a high concentration of glucose. The present invention also provides a method of producing brown adipocytes using pluripotent stem cells, which are useful for preparing the supernatant of brown adipocytes. The present invention has succeeded in obtaining a supernatant having a metabolism-improving action from brown adipocytes. It was also possible to obtain the supernatant without using a culture solution comprising a high concentration of glucose. The present invention has also succeeded in producing brown adipocytes from pluripotent stem cells using a feeder-free culture system without adding a cytokine cocktail or the like. The brown adipocyte supernatant of the present invention is expected to be applied to the development of therapeutic agents and cosmetic products for glucose metabolism diseases.

The present invention addresses the problem of inducing, from pluripotent stem cells, cells that are functional similar to PMC. Said problem is solved by a method that is for inducing peritoneal mesothelium cells from pluripotent stem cells and that comprises: a first culturing step for culturing pluripotent stem cells in the presence of a basic fibroblast growth factor and a bone morphogenetic factor; a second culturing step for culturing the cells cultured in the first culturing step, in the presence of the bone morphogenetic factor but at a concentration higher than that in the first culturing step, and the basic fibroblast growth factor; and a third culturing step for culturing the cells cultured in the second culturing step in the presence of a vitamin-A compound, a glycogen synthase kinase-3 inhibitor, and an inhibitor of activin kinase-2 and activin kinase-3.