1457results about "Non-embryonic pluripotent stem cells" patented technology

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized liver organoid and a construction method thereof. Aiming at the problems that the existing liver organ vascularization mostly adopts a co-culture mode, the operation process is complicated, the cell source heterogeneity is relatively high, the randomness is strong, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized liver organ. The preparation method comprises the following steps: sequentially inducing and culturing directional endoderm cells, posterior preintestinal cells and hepatic endoderm cells, adding an S1 culture medium containing VEGF to induce and culture for 1-3 days, then adding an S2 culture medium containing VEGF, BMP4 and FGF2 to induce and culture for 3-5 days, and then adding an S3 culture medium containing VEGF to induce and culture for 14-20 days to obtain the co-differentiated vascularized liver organ. According to the method disclosed by the invention, the operation process is simplified, the experiment efficiency is improved, the liver organ which is rich in blood vessel network, high in maturity and of a blood vessel-bile duct structure is obtained, and a foundation is laid for large-scale production and clinical application.

The invention relates to a method of modulating the expression of an endogenous gene in a somatic cell derived from an induced pluripotent stem cell (iPSC), comprising differentiating an engineered iPSC capable of stable or inducible expression of a CRISPR-related protein into a somatic cell prior to introduction of a guide RNA (gRNA) sequence. The invention also relates to the engineered iPSC, cells obtained by the method and uses thereof.

Embodiments of the invention include compositions and methods related to engineered human mucosal-associated invariant T (eMAIT) cells for off-the-shelf use for clinical therapy for cancer, infectious, and autoimmune diseases. In some embodiments, the eMAIT cells are produced from healthy human donor peripheral blood, cord blood, or G-CSF mobilized peripheral blood. In particular embodiments, the eMAIT cells are produced from a pluripotent stem cell line and therefore can be of unlimited supply. In some embodiments, the eMAIT cells are engineered to express chimeric antigen receptors (CARs), or / and immune regulatory molecules, or / and allorejection resistance molecules. Embodiments of the invention also include compositions of matter comprising polynucleotides encoding mucosal-associated invariant T cell receptor alpha chain polypeptides and / or mucosal-associated invariant T cell receptor beta chain polypeptides.

The invention relates to a cochlea spiral neuron in-vitro organ as well as a construction method and application thereof, and provides a method for obtaining a mature inner ear neuron by utilizing human pluripotent stem cell differentiation in vitro and further constructing an inner ear neuron in-vitro organ on the basis of the mature inner ear neuron. The internal ear neuron in-vitro organoid function stage obtained by the method and the application thereof are also provided.

Provided are: a method for producing an organoid, the method comprising a step for culturing somatic stem cells in a culture medium containing an agonist of the interferon-γ receptor; and a culture medium for producing an organoid, the culture medium containing an agonist of the interferon-γ receptor.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized islet organoid and a construction method thereof. Aiming at the problems that the existing pancreatic islet organ vascularization mostly adopts a co-culture mode, the operation process is long, the difficulty is high, the randomness is realized, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized pancreatic islet organ. The method comprises the following steps of: sequentially inducing and culturing directional endoderm cells, original intestinal canal cells and posterior preintestinal cells, and adding a culture medium containing VEGF (vascular endothelial growth factor), BMP4 (bone morphogenetic protein 4) and FGF2 (fibroblast growth factor 2) for culturing in subsequent induced differentiation, so as to obtain the co-differentiated vascularized islet organ. Under the condition that exogenous endothelial cells are not added, the iPS cells are directionally differentiated to generate the vascularized islet organ containing beta, alpha and delta cells and vascular endothelial cells for the first time, the vascularized islet organ is more mature than non-vascularized islet organs and has a stronger insulin secretion function, and a new thought and an experimental basis are provided for realizing functional healing of diabetes mellitus.

The invention discloses a construction method of a homogenized self-assembled heart organoid, and belongs to the field of organoid. The preparation scheme of the heart organoid provided by the invention is controllable in method and simple in procedure, emphasizes the transition of differentiation from an iPSC 2D level to a 3D self-assembly heart organoid, overcomes the heterogeneity, including size, structure, function and even gene difference, of cross-batch or even same-batch organoid caused by an existing differentiation method, and has good cross-batch reproducibility. The constructed heart organoid contains various cell types and can continuously beat for at least 180 days in vitro, and the heart organoid in the same batch is uniform in height and stable in function. The preparation scheme does not need an engineering scaffold, matrigel and the like, emphasizes self-assembly of heart organs, and better accords with physiological characteristics of a human body.

The invention discloses a sheep formation state pluripotent stem cell culture medium and application thereof. The invention belongs to the technical field of biology, and particularly relates to a sheep formation state pluripotent stem cell culture medium and application thereof. The composition for culturing the sheep formation state pluripotent stem cells, disclosed by the invention, contains an N2B27 basic culture medium and cell factors, wherein the cell factors contain CHIR99021, IWR1, A419259, a recovery hub IL6, a recovery hub sIL6R, a recovery hub Activin A and a recovery hub FGF2 (Fibroblast Growth Factor 2). The culture medium prepared from the composition can obtain stable sfPSCs, and can be applied to construction of sheep gene editing model animals.

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.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.

The invention provides a preparation method of ovarian stem cells and application of the ovarian stem cells in ovarian aging resistance. More specifically, ovarian stem cells and polypeptide LRP-C5 which is separated from pilose antler polypeptide and has a good ovarian aging treatment effect are prepared and screened from ovaries, ovarian aging of rats can be well treated after the ovarian stem cells and the polypeptide LRP-C5 are used independently or jointly, and the application prospect is good.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetically modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.

An object of the present invention is to provide a cell population suitable for transplant of retinal tissue and a method of production thereof. The present invention provides a cell population for transplant, comprising retinal cells with a modified bipolar cell-regulating gene and a method of production thereof.

The present invention relates to a method of transiently inducing aging in stem cells or somatic cells derived from induced pluripotent stem cells (iPSCs) by forward programming, the method comprising introducing one or more aging-inducing factors into the cell or exposing the cell to one or more exogenous aging-inducing factors for a period of time, the one or more aging-inducing factors are then removed or reduced, such as before or after forward programming of the iPSC, resulting in a reversal of the one or more aging phenotypes in the cell. Also provided are artificially aged stem cells or somatic cells derived from iPSC by forward programming, and their use in methods of identifying genes or combinations of genes involved in aged phenotype reversal or non-aged phenotype maintenance in cells. Also provided are methods of identifying a gene or combination of genes involved in aging phenotype reversal or non-aging phenotype maintenance in a cell comprising introducing one or more aging-inducing factors into the cell or exposing the cell to one or more exogenous aging-inducing factors for a period of time, and measuring the aging phenotype of the cell. In particular, methods of identifying a gene or combination of genes involved in aging phenotype reversal or non-aging phenotype maintenance in a cell optionally include loss of function, inhibition, knockout, function acquisition, or combination screening. Also provided is a method of regenerating somatic cells comprising altering the expression and / or activity of one or more genes identified by the methods described herein.

Provided are a RIC cell, and a preparation method therefor and application thereof. The present invention particularly relates to the lineage-specific differentiation of a pluripotent or multipotent stem cell, specifically to a RIC cell differentiated from a stem cell, and a preparation method therefor and application thereof.

The present invention relates to a modified stem cell and an ex vivo method for preparing defined mature cells from genetic programming of said modified stem cell which can be carried out in absence of certain growth factors, as well as a kit allowing to transform stem cells, and the different uses of the mature cells, for example for in vitro drug screening and toxicology or as cultivated meat. The genetic programming involves a molecular switch between two mutually exclusive programs of stem cell expansion and differentiation. The invention may be used for stem cells or other progenitors cells of any type, from any eukaryotic organism, but finds particular application in pluripotent stem cells from humans and livestock animals.

The invention provides a human pluripotent stem cell single cell cloning method and a culture medium thereof, and belongs to stem cell culture. A single cell is inoculated into a 96-hole or 384-hole culture plate by utilizing a limited dilution technology, and meanwhile, the proliferation process of the single cell is tracked and recorded in real time by virtue of a living cell analysis system, so that the obtained clone is ensured to be derived from the single cell. According to the method provided by the invention, a large number of clone groups derived from single cells can be simply, conveniently, quickly and easily obtained, and a solid foundation is laid for future large-scale production and industrial technical platforms.

The present invention discloses a method for enhancing cardiomyocyte production from induced pluripotent stem cells (iPSCs) by adjusting the culture medium exchange process during the transition from WNT activation to inhibition for differentiation. Instead of completely replacing the culture medium, this innovative technique involves partially changing the medium, which allows for a controlled exposure to WNT activators concurrent with the introduction of WNT inhibitors. This key modification in the protocol markedly increases the consistency and efficiency of cardiomyocyte generation, reducing variability between batches. By fine-tuning the medium exchange process, this method streamlines the creation of cardiomyocytes for use in regenerative medicine, emphasizing the critical adjustment phase within the culture conditions.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a Myo1B protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

The invention relates to the technical field of biology, in particular to a composition for preparing a brain organ, the brain organ and application of the brain organ. The invention further discloses a construction method of the brain organ, and on the basis of the method, different stem cells such as ESCs and iPSCs can be utilized to stably construct the three-dimensional brain organ model capable of simulating the medulla oblongata trigeminal nerve. The invention further discloses an organoid assembly which has important application potential in the fields of research on development, functions and related diseases of brain nuclei, drug discovery and the like.

The invention provides universally acceptable “off-the-shelf” hypoimmunogenic pluripotent cells and differentiated cardiac, endothelial, neuronal, islet, or retinal pigment cells thereof. Such hypoimmune cells are used to treat patients in need thereof. The cells lack major immune antigens that trigger immune responses and are engineered to avoid phagocytic endocytosis.

Materials, methods, and systems are provided for cell redifferentiation and expansion of induced pluripotent stem cell (iPSC)-derived hematopoietic stem cells (iHSCs) without the use of serum or additional cells.

The invention discloses a liver progenitor organoid culture medium and a culture method, and belongs to the technical field of organoid culture. The culture medium comprises a basic culture medium, Jagged-1, Dexamehysone, TGF (Transforming Growth Factor)-alpha, beta-mercaptoethanol, BMP4 (Bone Morphogenetic Protein 4), FGF2 (Fibroblast Growth Factor 2), specific addition factors Glutamax, HEPES (Hydroxyethyl PES), Nicotinamide, Penicillin and the like. The liver progenitor organoid is constructed by using the liver tissue for the first time, continuous growth and efficient proliferation of human liver progenitor cells in vitro can be promoted under the condition that gene editing is not carried out, meanwhile, the pluripotency of the liver progenitor organoid is maintained, and the consistency of specific marker expression is maintained; and a novel batch model is provided for the evaluation of hepatotoxicity at the present stage. From the long-term view, a foundation is laid for constructing a liver organ model with more advanced various differentiated cells.

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and / or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ / Activin / Nodal signaling and BMP signaling are provided. Also provided are methoc and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

The invention discloses a mouse lung organ and fibroblast co-culture method, which comprises the following steps of: performing in-vitro mouse lung organ culture on mouse lung tissue primary epithelial stem cell spheres (pneumospheroids) in a Transwell upper chamber, and after the lung organs in the Transwell upper chamber are differentiated, replacing with a lower chamber paved with mouse lung fibroblasts, therefore, a gas-liquid interface co-culture system of the mouse lung organ (upper chamber) and the fibroblast (lower chamber) is formed, and the hierarchical structure, morphological characteristics and regeneration function of the upper chamber organ and the phenotype, function and related signal path activation level of the lower chamber fibroblast can be dynamically researched.

The invention belongs to the field of biological medicines, and discloses a stress-induced mesenchymal stem cell extract and a preparation method and application thereof.The preparation method comprises the steps that S1, mesenchymal stem cells are cultured under the high-temperature condition, swelling and cracking are conducted on the mesenchymal stem cells, or supernate obtained after culture of the mesenchymal stem cells is taken and filtered, and filtrate is obtained; the temperature of the high temperature ranges from 37.0 DEG C to 42.0 DEG C; s2, purifying the mesenchymal stem cell extract by adopting a molecular sieve exclusion chromatography method or a reverse chromatography method. Specific high-temperature stimulation is adopted to induce the mesenchymal stem cells, a purification method is combined, protein expression is promoted, the purity of the mesenchymal stem cell extract is improved, the prepared and obtained purified Alibitumen is used in a cell oxidative damage model, and the cell oxidative damage rate is increased. It is found that all the Aribitua derived from high-temperature stress culture has high oxidative damage repairing capacity.