158 results about "Multipotential stem cell" patented technology

The invention relates to a new technology for promoting generation of hematopoietic precursor cells from human pluripotent stem cells. Specifically, the present invention relates to a method for preparing hematopoietic precursor cells comprising reducing the expression level of YTHDF2 gene in pluripotent stem cells or reducing the activity of YTHDF2 protein of pluripotent stem cells. On the other hand, the invention also relates to a kit for preparing the hematopoietic precursor cell, and the kit comprises a reagent for reducing the expression level of the YTHDF2 gene or a reagent for reducing the activity of the YTHDF2 protein.

The invention belongs to the field of stem cell culture, and relates to a pluripotent stem cell culture medium, which comprises an E6 basal culture medium, 2 ng / mL of TGF-beta1, 0-50 ng / mL of FGF-2, and 0.01-1 [mu] M of SUN11602. According to the technical scheme, proliferation and pluripotency of the hPSC are effectively maintained in a mode of combining small chemical molecules with low-concentration FGF-2.

This invention provides a method for propagating pluripotent stem cells (PSCs) by suspension culture in a bioreactor. [Solution] The method comprises the following steps: (i) adding a ROCK inhibitor (ROCKi) to pluripotent stem cells cultured in suspension in a bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding a sufficient excess volume of culture medium to reduce the concentration of the cell dissociation agent to a concentration at which cell aggregates can be re-formed; and (iv) culturing the mixture obtained in step (iii) under appropriate conditions that allow for the proliferation of PSCs.

Provided herein are compositions and methods for real-time identification and isolation of base-edited cell populations. Also provided herein are methods for producing enriched isogenic lines of genetically modified cells, including base-edited human pluripotent stem cells. In particular, provided herein are methods utilizing transient expression of reporter proteins, the detectable signal of which is altered following base editing. Using the transient reporter with a base editor permits enrichment of isogenic populations of base-edited cells.

Methods for generating mature somatostatin+ interneurons from human forebrain neural progenitor cells are provided using chemically-defined culture media in a two-stage culture protocol. The mature somatostatin+ interneurons are generated from medial ganglionic eminence neural progenitor cells (MGE-NPCs), which themselves are differentiated from pluripotent stem cells. Culture media, isolated cell populations and kits are also provided.

Disclosed are a method for producing a cell population in which regulatory T cells have proliferated, including (1) culturing a cell population containing CD4+ T cells derived from pluripotent stem cells in the presence of IL-4 and a TGF-βR agonist, a cell population containing regulatory T cells obtained according to the method, and a medicine containing the cell population containing regulatory T cells.

The present invention provides methods and compositions for the generation of microglial progenitor cells and microglial cells from pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells. The present invention also provides cells produced using such methods, and both methods of treatment and methods of drug screening that use such cells. Also provided are various tissue culture media, tissue culture media supplements, and kits useful for the generation of human microglial progenitor cells and human microglial cells.

The present disclosure provides methods of differentiating pluripotent stem cells, including inducible pluripotent stem cells, into lineage-specific midbrain floor progenitor cells, decisive dopaminergic neuron progenitor cells, definitive dopaminergic neuron progenitor cells, and / or dopaminergic neuron cells. Also provided are compositions use thereof, such as for the treatment of neurodegenerative diseases and conditions, including Parkinson's disease, as well as articles of manufacture and kits for use thereof.

The invention discloses an analysis method for storage of single vesicles in ventral spinal cord motor neuron-like organs induced and differentiated by an iPSC technology. The method sequentially comprises the steps of construction of an acetylcholine nano microelectrode sensor interface, recovery and culture of human pluripotent stem cells, directional differentiation of spinal cord movement organs and storage and detection of single vesicle acetylcholine. A new thought and a new method are provided for monitoring the level of acetylcholine stored in the single vesicles at the single cell level, and the method has important theoretical significance and wide practical value in research of pathogenesis of diseases, drug screening and the like in the field of neuroscience.

The invention relates to the technical field of biology, in particular to a method for efficiently obtaining human definitive endoderm cells in vitro, which comprises the following steps: S1, inoculating human pluripotent stem cells into matrigel, and performing maintenance culture for 16-24 hours by using a stem cell maintenance culture medium; s2, from Day-2 to Day-1, absorbing and abandoning the stem cell maintenance culture medium, adding a pretreatment culture medium for culture, and changing the pretreatment culture medium once every day; the pretreatment culture medium is a stem cell maintenance culture medium added with dimethyl sulfoxide; s3, carrying out Day0 to Day2, absorbing and abandoning the pretreatment culture medium, adding a definitive endoderm differential culture medium, carrying out induced culture, and changing the definitive endoderm differential culture medium once a day; the definitive endoderm differential culture medium is an RPMI1640 culture medium added with fetal calf serum and activin A; s4, Day3, obtaining the required human definitive endoderm cells, and solving the problems that the existing definitive endoderm differentiation efficiency and cell population purity are limited and are influenced by batch-to-batch variation.

The application belongs to the technical field of biological medicine, and provides a CXCR4 high expression type iPSC-NK cell with enhanced bone marrow and tumor tissue homing ability, and a preparation method and application thereof. The cell takes pluripotent stem cells as starting cells, overexpresses a membrane-bound IL-15 and IL-15RA fusion protein gene and a CXCR4 receptor gene in the pluripotent stem cells; pluripotent stem cells stably expressing the target gene are obtained, and then iPSC-NK cells are obtained through induction differentiation. The membrane-bound IL-15 and IL-15RA fusion protein gene and the CXCR4 receptor gene are targetedly integrated into a safe harbor site of the induced pluripotent stem cell through a gene editing technology, so as to construct an iPSC cell strain stably expressing key proteins; the function-enhanced iNK cell is obtained through induction differentiation, and exhibits excellent bone marrow and various solid tumor tissue homing ability and persistent immune killing activity.

The invention provides an induced pluripotent stem cell culture medium and a culture method of induced pluripotent stem cells. The induced pluripotent stem cell culture medium comprises a basic culture medium, a composite additive A and a composite additive B, the composite additive A comprises 5-30 [mu] g / mL of recombinant human insulin growth factors, 1-5 ng / mL of recombinant human transforming growth factors, 50-200 ng / mL of recombinant human basic fibroblast growth factors, 7-20 [mu] g / mL of sodium selenite, 0.5-2 mg / mL of albumin, 50-70 [mu] g / mL of ascorbic acid and 5-20 [mu] g / mL of transferrin; the composite additive B is prepared from 50 to 300mg / mL of a Stock B solution, 0.5 to 5 percent (v / v) of a non-essential amino acid solution, 50 to 200ng / mL of DL-piperidinecarboxylic acid, 50 to 200mu g / mL of gamma-aminobutyric acid, 0.1 to 1 percent (v / v) of a lipid concentrated solution, 0.5 to 5 percent (v / v) of an L-alanyl-L-glutamine solution and 0.1 to 0.1 percent of mercaptoethanol. The culture medium for the induced pluripotent stem cells provided by the invention can ensure the self-renewal capability of the iPSCs in a long-term passage process; and the genetic stability of the iPSCs cells is optimized according to the genome integrity and pluripotent state.

The invention relates to macrophages derived from induced pluripotent stem cells as well as a preparation method and application of the macrophages. Specifically, the present invention relates to a macrophage (iPSC-CAR-M) derived from an induced pluripotent stem cell (iPSC), in which the macrophage expresses a chimeric antigen receptor (CAR) that specifically binds to a tumor-associated antigen; and a transcription factor combination comprising PU.1, IRF5 and BATF2 is overexpressed. The invention also relates to a pharmaceutical composition containing the macrophage derived from the induced pluripotent stem cells, and a preparation method and application of the pharmaceutical composition.

This useful solution, in the field of biotechnology and regenerative medicine, relates to a process for enhancing the efficiency of induced pluripotent stem cell (iPSC) colony formation from human peripheral blood mononuclear cells (PBMCs). The solution aims to overcome the low reprogramming efficiency, unstable colony formation, and limited iPSC cloning rates commonly encountered in PBMC iPSC generation processes. The technical essence of the solution lies in the synchronized optimization of the reprogramming process, including pre-activating PBMCs with an appropriate cytokine combination at predetermined times before reprogramming, using a non-integrated Sendai virus system under optimized transformation conditions, switching culture media at defined intervals to promote cell state transitions, and applying a selection, enrichment, and stabilization process for iPSC cloning in the initial passages after colony formation.The process also incorporates cell line quality control steps through assessment of residual Sendai virus RNA removal and mycoplasma contamination testing during line maintenance. Experimental results show that the proposed process increases the number of iPSC colonies formed, improves the rate of establishing stable iPSC lines, and enhances cell homogeneity and pluripotency maintenance. The solution can be applied in basic research, pathogenesis modeling, drug screening, regenerative medicine, and studies using induced pluripotent stem cells derived from human peripheral blood cells.

The application belongs to the technical field of cell culture and organoid, and particularly relates to a culture medium and a culture method for inducing primitive pluripotent stem cells into amnion-like cells. The application provides a culture medium for inducing primitive pluripotent stem cells into amnion-like cells, and through accurate regulation of key signal pathways such as TGF-beta, Wnt and FGF / ERK, efficient and specific directional differentiation of primitive pluripotent stem cells into amnion-like cells can be achieved. The application has the advantages of clear composition and good repeatability, avoids the use of uncertain components such as serum, and provides a reliable tool for stably obtaining high-quality amnion-like cells. The cells produced thereby are an ideal starting point for constructing amnion organoids and isolating amnion mesenchymal stem cells, and lay an important technical foundation for embryonic development research, disease modeling and regenerative medicine applications.

[Problem] The first problem addressed by the present invention is to specify a previously unknown factor or a combination of previously unknown factors among factors or combinations thereof capable of efficiently producing egg-like cells obtained from pluripotent stem cells or the like. The second problem addressed by the present invention is to provide an egg-like cell and a method for efficiently producing same. [Solution] This method for producing an egg-like cell involves a step for introducing a factor into a cell, wherein the factor includes DLX6 or a nucleic acid encoding DLX6. This egg-like cell is derived from a human pluripotent stem cell and expresses PADI6.

The present invention provides a method for producing dopaminergic neuron progenitor cells from pluripotent stem cells, which method comprises the steps of: (i) performing adherent culture of pluripotent stem cells on an extracellular matrix in a medium containing a reagent(s) selected from the group consisting of BMP inhibitor, TGFβ inhibitor, SHH signal-stimulating agent, FGF8, and GSK3β inhibitor; (ii) collecting Corin- and / or Lrtm1-positive cells from the cells obtained in Step (i) using a substance which binds to Corin and / or a substance which binds to Lrtm1; and (iii) performing suspension culture of the cells obtained in Step (ii) in a medium containing a neurotrophic factor.

The application discloses a culture solution for indirectly inducing human induced pluripotent stem cells into mesenchymal stem cells, which is composed of a neuroectoderm induction culture solution, a mesenchymal stem cell differentiation culture solution and a mesenchymal stem cell maintenance culture solution. The neuroectoderm induction culture solution is composed of a Neurobasal culture medium, a DMEM / F12 culture medium, an N2 additive, a B27 additive, a GSK3 inhibitor and a TGF-beta inhibitor. The application further discloses a method for indirectly inducing human induced pluripotent stem cells into mesenchymal stem cells, which comprises the following steps: taking the induced pluripotent stem cells, culturing the induced pluripotent stem cells in the neuroectoderm induction culture solution for 6 days, then culturing the induced pluripotent stem cells in the mesenchymal stem cell differentiation culture solution for 7 days, and then culturing the induced pluripotent stem cells in the mesenchymal stem cell maintenance culture solution for 3-5 generations. The method can direct the induced pluripotent stem cells to differentiate into mesenchymal stem cells in a short time, and greatly shortens the culture time.

Tissue-specific progenitor cells representing the endoderm, ectoderm, and mesoderm lineages are generated from pluripotent sources. Progenitor cell generation is accomplished by contacting pluripotent stem cells with tissue-specific exosomes, microRNAs, proteins, and peptides, obtained by stressing the tissue under in vitro conditions. In vitro-generated tissue organoids are used as a source of "differentiation factors" for generating individualized progenitor cells. For example, to generate lung progenitor cells, allogeneic stem cells are implanted into decellularized cadaveric lung tissue to form lung structures, which are then exposed to various cellular stressors and conditioned media to differentiate the pluripotent stem cells into progenitor cells. Extracellular matrix can be obtained from decellularized structures seeded with regenerating cells and used to generate organoids, which are then subjected to stress to produce differentiation factors.

This invention provides a human embryonic stem cell expansion culture medium, comprising DMEM / F12 medium, ITS-A, HEPES, magnesium ascorbate phosphate, non-essential amino acids, bFGF, FGF2-G3, GlutaMAX, LY-333531, gentamicin sulfate, Y27632, and CHIR99021. Y-27632 inhibits pluripotent stem cell apoptosis, improves adhesion efficiency, maintains pluripotency and colony formation ability; CHIR99021 enhances the self-renewal of pluripotent stem cells and inhibits their differentiation; LY-333531 inhibits spontaneous differentiation of pluripotent stem cells; and gentamicin sulfate reduces the risk of contamination during culture. The culture medium provided by this invention is serum-free, free of allogeneic substances, has a simple composition, and offers excellent expansion speed, effectively maintaining long-term stable passage of human embryonic stem cells, providing stable technical support for scientific research.

A fish liver decellularized extracellular matrix based microfluidic 3D printing hydrogel for liver regeneration, and a preparation method thereof are provided. A fish liver decellularized extracellular matrix (dECM) is combined with gelatin methacryloyl (GelMA), and loaded with hepatic spheroids derived from induced pluripotent stem cells (iPSC-hep) for liver regeneration. The fish liver decellularized extracellular matrix based microfluidic 3D printing hydrogel of the present disclosure has excellent biocompatibility and retains intact endogenous growth factors, maintains the biological activity of cells, ensures effective cell encapsulation, and is conducive to robust functional expression of iPSC-hep. After being transplanted in vivo, the hydrogel significantly improves the survival rate and liver function of mice with acute liver failure, and promotes liver regeneration and repair.

The technical problem addressed by the present invention is to provide a method for producing an apparent cross-karyocyte-like organ. This technical problem is solved by a method for producing a cross-seeded organ, said method comprising: (1) a step for culturing pluripotent stem cells in an intermediate medium between an undifferentiated maintenance medium and a neural differentiation medium; (2) a step for culturing the cells obtained in step (1) in a neural differentiation medium; and (3) a step for forming a cell mass prior to the start of step (2): (A) 6000 or more raw material cells are inoculated to form one cell mass, and (B) when the start of step (2) is set to the 0th day, the culture medium contains a Sonic Hedgehog signaling pathway acting substance from the 3rd day, and when the start of step (2) is set to the 0th day, the culture medium contains the Sonic Hedgehog signaling pathway acting substance from the 3rd day.

The invention provides a low-immunogenicity pluripotent stem cell for expressing KRT7 and a preparation method of the low-immunogenicity pluripotent stem cell. The pluripotent stem cell comprises: a reduced MHC-I function compared to a parent pluripotent stem cell; a reduced MHC-II function compared to the parental pluripotent stem cell; and an increased expression of the KRT7 protein compared to the parental pluripotent stem cell. The low-immunogenicity pluripotent cell can significantly reduce or escape the recognition and attack of an immune system, especially the attack of T cells, natural killer cells, macrophages and the like, without changing the updating and differentiation characteristics of stem cells.

Tests regarding chromosomal abnormalities in pluripotent stem cells have involved cellular invasion, and have been unable to be applied to cells that require non-invasive testing, such as cells being cultured. Provided is an information processing system including: an image data acquisition unit configured to acquire image data including a colony of pluripotent stem cells; a first information acquisition unit configured to acquire, based on analysis of the image data, first information that is information regarding a predetermined morphology of the colony of pluripotent stem cells; and a second information generation unit configured to generate, based on the first information, second information that is information regarding chromosomal abnormalities in the pluripotent stem cells.

The invention discloses a stem cell-loaded microcapsule, a preparation method and application, the stem cell-loaded microcapsule is composed of hydrogel and stem cells, and the stem cells are wrapped in the stem cell-loaded microcapsule; the stem cells are selected from at least one of umbilical cord mesenchymal stem cells, bone marrow mesenchymal stem cells, adipose-derived stem cells, embryonic stem cells and induced pluripotent stem cells; the number of living cells in the stem cell-loaded microcapsule is 1-10; the particle size of the stem cell-loaded microcapsule is 30-200m, and the dispersion coefficient of particle size distribution is 0.01-20%. According to the preparation method, stem cells and a biological material are mixed to prepare a prepolymerization solution, the biological material provides immune protection for the cells, the in-vivo retention time of the cells is prolonged, and the problem that at present, the stem cells are rapidly removed by an immune system after being transplanted into the body is solved. The capsule can be used as bio-ink to be assembled into a bionic stent through 3D printing or directly injected to an injured part, and has a wide application prospect.

The invention discloses a stem cell microcapsule-entrapped tissue adhesive, a preparation method and application, the tissue adhesive is composed of stem cell microcapsule-entrapped hydrogel and a biological adhesive, and live stem cells are entrapped in the tissue adhesive; the stem cells are selected from at least one of umbilical cord mesenchymal stem cells, bone marrow mesenchymal stem cells, adipose-derived stem cells, embryonic stem cells and induced pluripotent stem cells; the biological adhesive is formed by compounding a hydrophilic polymer and a block polymer, wherein the mass ratio of the hydrophilic polymer to the block polymer is (1: 5)-(5: 1); the transmittance of the tissue adhesive is 60-100%, the tissue adhesive strength is 20-200 kPa, and the volume swelling ratio is smaller than or equal to 50%. According to the scheme, the stem cell entrapped microcapsule and the biological adhesive are mixed to prepare the tissue adhesive, and the adhesive can be used as bio-ink to be assembled into a bionic scaffold through 3D printing or directly injected to an injured part, so that the tissue adhesive has a wide application prospect.

To provide a method and composition for generating induced non-pluripotent cells differentiated from single-cell induced iPSC (induced pluripotent stem cell) clone lines. [Solution] A method and composition are provided for obtaining functionally enhanced induced effector cells obtained from targeted differentiation of genome-manipulated iPSCs. The iPSC-induced cells provided herein have stable functional genome editing that results in improved or enhanced therapeutic effects. Therapeutic compositions and their use are also provided, comprising functionally enhanced induced effector cells alone or in combination therapy with antibodies or checkpoint inhibitors.

A population of universal natural killer (NK) cells derived from human pluripotent stem cells (hPSCs) and engineered to overexpress the transcription factor ID2, NFIL3, and / or SPI1 and, optionally, an anti-programmed death ligand 1 (PD-L1) chimeric antigen receptor (CAR) and an anti-fluorescein isothiocyanate CAR are provided. Methods of treating cancer in a subject using the population of universal NK cells are also provided.

Disclosed herein are methods for obtaining endometrial stromal fibroblast cells from pluripotent stem cells, such as induced pluripotent stem cells. The present disclosure also provides methods of obtaining a three-dimensional, multilayered endometrial tissue composition. Methods of using the cells and tissue compositions in drug screening and therapeutic applications are also provided.