295 results about "Pluripotential stem cell" patented technology

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing intestinal organs and application of the reagent combination or kit. According to the reagent combination or the kit, intestinal organs can be obtained from cell-derived epithelial cells obtained from a donor in a non-invasive manner, and the obtained intestinal organs can be cryopreserved and recovered and can be amplified in vitro for a long time; transcriptome characteristics are similar to those of real human small intestine tissues, and typical marker genes of various small intestine pedigree cell types are highly expressed; compared with intestinal organs obtained through induction of pluripotent stem cells, the intestinal organs have more intestinal pedigree characteristics, and the intestinal function development is more mature; after being promoted to be mature, the intestinal organ also highly expresses genes related to drug absorption and metabolism, has drug absorption capability similar to that of an immortalized intestinal cell line, but more prominently shows intestinal cell lineage characteristics, is closer to an intestinal environment in a real human body, and can be used for screening intestinal disease drugs; the intestinal barrier function is realized.

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.

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 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 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 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 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.

The present invention generally relates to compositions and methods useful for treating a brain injury such as stroke, optic neuropathy, traumatic brain injury, and cerebral palsy. The methods include administering HMCs obtained by in vitro differentiation of pluripotent stem cells and / or extracellular vesicles (EVs) derived from such HMCs (HMC-EVs) into a subject.

The present invention provides methods of constructing vascularized retinal organoids by differentiating retinal progenitor cells and endothelial cells. The method is co-cultured by differentiating retinal progenitor cells and endothelial cells, and comprises the following steps: carrying out multiplication culture on pluripotent stem cells until the cell confluence degree is 80-90%, and digesting; carrying out differentiation culture on the digested pluripotent stem cells in a culture medium containing retina induction factors to obtain retina progenitor cells; and co-culturing the retinal progenitor cells differentiated for 12-13 days and endothelial cells to obtain the vascularized retinal organ, wherein the endothelial cells are obtained by any one method of induced differentiation of digested pluripotent stem cells or culture of endothelial cell lines. The vascularized retinal organ with the retinal layered structure is constructed by integrating the endothelial cells and the retinal progenitor cells for the first time, and the limitation that in the prior art, the retinal organ lacks vascularization is successfully overcome.

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 present invention provides a method of producing a population of mesenchymal stromal cells (MSC) of CD73 + CD44 +, CD90 +. The CD73 + CD44 +, CD90 + MSC is used in a method for producing terminally differentiated osteoblasts, adipoblasts and chondroblasts from pluripotent stem cells (PSCs). The differentiation methods include the use of a single agent-a WNT signaling pathway activator used on an adherent culture of PSC, such as a GSK3 [beta] inhibitor.

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.

The present disclosure provides efficient in vitro methods for producing megakaryocytes and / or platelets from human induced pluripotent stem cells (iPSCs). In various embodiments, the cells and platelets produced in accordance with the present disclosure are functional and / or more closely similar to corresponding lineages isolated from peripheral blood, bone marrow, or other tissue. The present invention provides, in some aspects, isolated cells / platelets and compositions produced by the methods disclosed herein, as well as methods for use in therapy.

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 invention belongs to the technical field of regenerative medicine, and particularly relates to a method for generating vascular endothelial cells by inducing pluripotent stem cells to differentiate in vitro. The method comprises the following steps: carrying out first culture on pluripotent stem cells in a differential medium I to obtain cells A; carrying out second culture on the cell A in a differential culture medium II, and differentiating and directionally forming an endoderm cell B; performing third culture on the endoderm cells B in a differential culture medium III to obtain differentiated cells C; carrying out fourth culture on the differentiated cells C in a differentiation culture medium IV, and differentiating to form differentiated cells D; carrying out fifth culture on the differentiated cells D in a differential culture medium V, and differentiating to obtain vascular endothelial cells; wherein the differential culture media I-V do not contain activin A, VEGF (vascular endothelial growth factor) and BMP4 (bone morphogenetic protein 4). The method can improve the differentiation efficiency of the IPS cells to the vascular endothelial cells, and has a wide prospect in the cell therapy industry.

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.

The purpose of the present invention is to provide a novel medical application using pluripotent stem cells in regenerative medicine. The present invention provides a cell formulation and pharmaceutical composition that are for amelioration and therapy for frequent urination and bladder pain caused by the inflammation of the bladder and that contain SSEA-3 positive pluripotent stem cells isolated from cultured mesenchymal cells or a mesenchymal tissue of a living organism. The cell formulation according to the present invention is considered to be based on a mechanism in which, for example, Muse cells are administered against interstitial cystitis having the abovementioned disorders to be successfully engrafted into affected bladder tissue, whereby the disorders are ameliorated and treated.

The present disclosure provides, in various aspects and embodiments, methods for generating hematopoietic lineages for cell therapy, including erythroid progenitor cells, progenitor cell erythroblasts, granulocyte-macrophage progenitor cells (GMP), and megakaryocyte erythroid progenitor cells (MEPs) and erythroid cells. In various embodiments, the invention provides efficient in vitro methods for developing such hematopoietic lineages, including but not limited to progenitor erythroblasts and erythroblasts lineages, from human induced pluripotent stem cells (iPSCs). The cells produced in various embodiments according to the present disclosure are functional and / or more closely similar to corresponding lineages isolated from peripheral blood or bone marrow. The invention also provides isolated cells and cell compositions produced by the methods disclosed herein, as well as methods for cell therapy.

The invention discloses a GLP-1 (32-36) engineered exosome and application of the GLP-1 (32-36) engineered exosome in drugs and instruments for preventing and / or treating diabetic lower limb arterial lesion. The preparation process and related characteristics of the GLP-1 (32-36) engineered exosome are respectively as shown in figures (1) and (2). The exosome is prepared by treating human pluripotent stem cells by using GLP-1 (32-36). The GLP-1 (32-36) engineered exosome has relatively high activity in plasma, and the enterotrypsin synthetic peptide can effectively improve the yield of the exosome and further optimize the function of the exosome, so that a stronger therapeutic effect of promoting angiogenesis after ischemia is exerted. The GLP-1 (32-36) engineered exosome disclosed by the invention also has safety and can be used for a treatment strategy of diabetic lower limb artery complications.

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 present invention relates to a composition for promoting the efficiency of somatic cell replication, comprising endothelin as an active ingredient. When somatic cells are treated with the composition of the present invention, H3K9me3, which is suspected to be a factor for the lowering success rate of nuclear substitution, is inhibited, and H3K4me3, which has the opposite effect, is increased to increase the success rate of nuclear substitution. Accordingly, the composition of the present invention increases the efficiency of somatic cell replication, thereby increasing the efficiency of producing pluripotent stem cells derived from nuclear substitution, and ultimately, the composition is expected to contribute to the development of cell therapeutic agents.

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.