120 results about "Multipotent Stem Cell" patented technology

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.

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 present invention provides a method for producing a retinal progenitor cell, including (1) a first step of subjecting pluripotent stem cells to floating culture in a serum-free medium to form an aggregate of pluripotent stem cells, and (2) a second step of subjecting the aggregate formed in step (1) to floating culture in a serum-free medium or serum-containing medium each being free of a substance acting on the Sonic hedgehog signal transduction pathway but containing a substance acting on the BMP signal transduction pathway, thereby obtaining an aggregate containing retinal progenitor cells.

The invention belongs to the technical field of biomedicine, and relates to a method for increasing the yield of pancreas islet organs differentiated by pluripotent stem cells and improving the stability. The method comprises the following steps: (1) inoculating pluripotent stem cells, and culturing by adopting a basic culture medium containing Y-27632; (2) changing the liquid, and continuously culturing by adopting a basic culture medium; (3) culturing for 1 day by using an S1D1 culture medium, and then culturing for 2-3 days by using an S1D2 culture medium; (4) changing the liquid, and culturing for 2-3 days by adopting the S2 culture medium; (5) changing the liquid, and culturing for 1-2 days by adopting an S3 culture medium; (6) changing the liquid, and culturing for 4-7 days by adopting a culture medium S4; (7) changing the liquid, and culturing for 3-10 days by adopting an S5 culture medium; and (8) carrying out cell sphere repolymerization, and culturing for 3-21 days by using an S6 culture medium to obtain the pancreas islet organ. The present invention increases the yield and differentiation efficiency at each differentiation stage and increases the scale yield at the cellular level.

The present invention aims to provide a method for efficiently producing a cell mass containing pituitary tissue from pluripotent stem cells. A method for producing a cell mass containing pituitary tissue, including the following steps (1) and (2):(1) a first step of suspension-culturing pluripotent stem cells to form a cell aggregate in the presence of a Wnt signal transduction pathway inhibiting substance,(2) a second step of suspension-culturing the aggregate obtained in the first step in the presence of a BMP signal transduction pathway activating substance and a Sonic hedgehog signal transduction pathway activating substance, thereby obtaining a cell mass comprising pituitary tissue.

The present application relates to a kind of RIC cells and its preparation method and application, especially it is related to the lineage-specific differentiation of pluripotent or multipotent stem cells, specifically related to a kind of RIC cell obtained from stem cell differentiation and its preparation method and application.

Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells and a culture medium composition thereof. Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells or NK cells and a culture medium composition thereof. The NK cells obtained using the provided culture method have high purity and good in vitro expansion effect; the yield of iNK cells is high, and a single iPSC can differentiate into approximately 2,000 NK cells; and the obtained iNK cells highly express CD16 (over 70%). As a result, the problem of low CD16 expression in iNK cells in the prior art, which requires genetic modification methods to solve, has been addressed.

The present disclosure relates to genetically modified cells (e.g., iPSC, IPS-derived immune cells, e.g., NK or T cells, or NK cells or T cells) comprising a disrupted FAS gene, an insertion of a polynucleotide encoding an IL15 / IL15Rα fusion, and an insertion of a polynucleotide encoding a CAR, e.g., an anti-GPR87 CAR. Therapeutic uses of the genetically modified cells to treat cancer (e.g., a lung cancer) are also provided.

The utility model discloses an induced pluripotent stem cell clone selection needle head which comprises a needle head body. The needle head body comprises a first straight needle body, a bent needle body, a second straight needle body and a needle tip; one end of the first needle body is connected with a needle cylinder, and the other end of the first needle body is connected with one end of the bent needle body; the other end of the bent needle body is connected with one end of the second needle body; the other end of the second needle body is connected with the needle tip; the included angle between the second needle body and the first needle body is an acute angle. According to the utility model, the technical problem of low efficiency caused by low clone selection accuracy is solved, and the technical effects of accurate clone selection and efficiency improvement are achieved.

The present invention provides a novel method for producing natural killer (NK) cells, which enables production of a large amount of high-quality NK cells easily, inexpensively, and safely in a short period of time from pluripotent stem cells. Specifically, a method for producing NK cells is provided, including(1) a step of forming pluripotent stem cell spheres having an average particle size of not less than 200 μm in a first medium;(2) a step of inducing the pluripotent stem cell spheres formed in step (1) into a cell population including hematopoietic progenitor cells by three-dimensional culture using a second medium; and(3) a step of inducing the cell population including hematopoietic progenitor cells obtained in step (2) into a cell population including NK cells by three-dimensional culture using a third medium,wherein the steps (1) to (3) are performed by a perfusion culture method.

A method for assessing differentiation potential of cells in a culture broth in the differentiation of pluripotent stem cells into neural cells of the midbrain floor plate region, comprising: measuring a concentration of NT-3 in a culture supernatant in a culture broth obtained by culturing the pluripotent stem cells in a medium containing an inducer of differentiation into the neural cells of the midbrain floor plate region; comparing the measured concentration of NT-3 with a reference concentration; and when the concentration of NT-3 is equal to or more than the reference concentration, assessing the cells in the culture broth as capable of differentiating into the neural cells of the midbrain floor plate region, wherein the culture supernatant is a culture supernatant collected from the culture broth at any time from 48 hours to 240 hours after the start of culture of the pluripotent stem cells.

Provided is a method for inducing T cells for a cell-based immunotherapy, comprising the steps of:(1) providing Rag 1 and / or Rag 2 gene knockout human pluripotent stem cells bearing genes encoding a T cell receptor specific for a desired antigen, and(2) inducing T cells from the pluripotent stem cells of step (1). Further provided are a cell-based immunotherapy method that uses the T cells for the cell-based immunotherapy and an iPS cell bank for the cell-based immunotherapy.

The present disclosure provides methods for generating hematopoietic progenitor cells. In some embodiments, the methods involve an in vitro or ex vivo cell culture model utilizing rentionic acid signaling for producing hematopoietic progenitor cells from pluripotent stem cells.

The present invention provides a method for producing neural crest cells from pluripotent stem cells, the method comprising: a step (1) of subjecting pluripotent stem cells to suspension culture in a culture medium containing an ALK inhibitor and a bone morphogenetic protein; and (2) performing suspension culture in a culture medium containing an ALK inhibitor and a GSK-3beta inhibitor.

The invention belongs to a cell culture technology in the field of biomedicine, and particularly relates to a liver organoid for traditional Chinese medicine screening and toxicity assessment and a construction method and application thereof. The construction method comprises the following steps: carrying out plane multiplication culture on pluripotent stem cells, then digesting and subculturing, and repeating to obtain a cell culture medium; a pluripotent stem cell group which is uniform in cell morphology and free of spontaneous differentiation is obtained; performing two-dimensional plane directional induction on the pluripotent stem cell population to enable cells to synchronously and uniformly receive differentiation signals to obtain hepatogenic mother cells; culturing the obtained hepatic mastocytes by using a culture medium E under a three-dimensional suspension condition to promote the specificity and balling of the hepatic lineage; the culture medium E is an Advanced DMEM / F12 which comprises 1 to 5 [mu] M of RA, 2 to 10 [mu] M of Y-27632, 0.5 to 1 [mu] M of beta-nicotinamide mononucleotide, 5 to 10 [mu] M of senkyunolide A, B27 and N2; and then replacing the culture medium to further mature and functionalize the hepatocytes to obtain the liver organoid, and the obtained liver organoid has high stability, high uniformity, good survival rate and mature hepatocyte functions.

The present invention relates to the field of stem cell biology, in particular the linage specific differentiation of pluripotent or multipotent stem cells, which can include, but is not limited to, human embryonic stem cells (hESC), human induced pluripotent stem cells (hiPSC), somatic stem cells, cancer stem cells, or any other cell capable of lineage specific differentiation. Specifically described are methods to direct the lineage specific differentiation of hESC and / or hiPSC to nociceptors (i.e. nociceptor cells) using novel culture conditions. The nociceptors made using the methods of the present invention are further contemplated for various uses including, but limited to, use in in vitro drug discovery assays, pain research, and as a therapeutic to reverse disease of, or damage to, the peripheral nervous system (PNS). Further, compositions and methods are provided for producing melanocytes from human pluripotent stem cells for use in disease modeling.

Methods provided herein describe a way to generate NK cells from pluripotent stem cells by disrupting the expression of a gene.

To provide methods for producing an artificial skeletal muscle tissue from pluripotent stem cells.SOLUTION: The present invention describes a method for preparing an engineered skeletal muscle tissue, as well as skeletal myoblasts, skeletal myotubes, and satellite cells, a medium used being serum-free and different chemicals and their concentrations, as well as the physical stimuli, being defined. Additionally, the methods described herein are performed without transfection of human cells with transgenes. The engineered skeletal muscle cells exhibit myoblast-, myotube-, or satellite cell-specific genetic markers, demonstrating efficient differentiation of these cell types. Despite its engineered production, the skeletal muscle tissue possesses excellent stimulus-dependent contractility and exhibits contractions in response to different stimulation frequencies.SELECTED DRAWING: None

The present invention discloses a method for detecting the activity of islet cells derived from human pluripotent stem cells: the islet cell spheres to be detected are digested to obtain a single-cell suspension; 0.1-5% HSA by volume is added to the PBS buffer as a cell protection solution, the cell protection solution is added to the single-cell suspension, and then PI staining solution is added for staining; the stained sample is subjected to flow cytometry to obtain the proportion data of live cells. The present invention uses a low-concentration PI staining solution for staining to improve the staining effect. The stained cells are sorted by flow cytometry. HSA is used as a cell protection solution to protect the islet cells during the flow cytometry detection, reduce the sample preparation time and the damage to islet cells caused by flow cytometry detection, maintain cell activity, can be standardized and normalized, avoid human judgment errors, objectively and accurately detect the islet viability, has high repeatability, and has a low detection cost, and is expected to be applied to the determination of viability before clinical islet transplantation.

Provided is a method for differentiating pluripotent stem cells in vitro to prepare a cell population mainly composed of midbrain floor plate dopaminergic progenitor cells, wherein the progenitor cells can efficiently and directionally differentiate into midbrain dopaminergic neuron precursors and midbrain dopaminergic neurons after being transplanted into a subject. Further provided is a cell population prepared by the method, a pharmaceutical composition containing the cell population, and a method for using the cell population or the pharmaceutical composition to treat or ameliorate a neurological disorder caused by a reduction in the number of midbrain dopaminergic neurons and / or impairment of function thereof.

Compositions and methods are provided for modifying eukaryotic cells, e.g., to express a transgene of interest and / or to generate a population of expanded cells ex vivo.SOLUTION: A method of transducing a population of eukaryotic cells, such as a population of pluripotent cells, to express a gene of interest by contacting the cells with a viral vector, e.g., a lentiviral vector, and a diblock copolymer, e.g., a diblock copolymer comprised of a hydrophilic region and a hydrophobic region. For example, the diblock copolymer can be composed of polyoxyethylene (PEO) subunits and polyoxypropylene (PRO) subunits. In addition, the compositions and methods of the invention can be used to promote the ex vivo expansion or survival of a population of pluripotent cells (e.g., CD34 + hematopoietic stem or progenitor cells), for example, by contacting the cells with a diblock copolymer.SELECTED DRAWING: None

This invention provides a primer-probe composition, kit, and application for detecting iPSC residues, relating to the field of pluripotent stem cell residue detection technology. The biomarkers include the ZIC3, ADAM19, ADD2, or ZSCAN10 genes, solving the technical problem of the lack of marker genes in existing technologies capable of detecting or assessing iPSC residues in different tissues and functional cells derived from iPSCs. The primer-probe combination of biomarkers can specifically amplify the ZIC3, ADAM19, ADD2, and ZSCAN10 genes respectively, enabling quantitative detection of each biomarker in cells. This improves the accuracy of iPSC residue detection or assessment results, ensures the safety of iPSC-derived cells, and reduces the clinical risks of related applications. The method is time-efficient, convenient, rapid, and highly sensitive, capable of quantitatively analyzing the proportion of iPSC residues in samples, ensuring product safety.

The invention provides a method of producing a population of CD73+ CD44+, CD90+ mesenchymal stromal cells (MSCs). The CD73+ CD44+, CD90+ MSCs are used in methods of generating terminally differentiated osteogenic, adipogenic and chondrogenic cells from pluripotent stem cells (PSCs). The differentiation method includes the use of a single agent, WNT signaling pathway activator such as a GSK3β inhibitor, used on adherent culture of PSCs.

Methods comprise exposing a culture of human pluripotent stem cells adherent on a first substrate comprising a first laminin to a differentiation medium for a first time period of 15 days to 50 days to obtain a first population of adherent cells comprising RPE cells and / or progenitors thereof; at the end of the first time period, dissociating the first population from the first substrate; replating the dissociated first population of cells on a second substrate comprising a second laminin; and culturing the replated first population of cells on the second substrate for a second time period to obtain an expanded and matured second population of cells comprising the RPE cells. The first and second laminins are independently selected from LN-521, LN-511, LN-111 and LN-121, and are an intact protein or protein fragment. Also included are RPE cells and materials and compositions utilizing such RPE cells for various treatments.

Methods for producing CD31 + VE-cadherin + KDR + endothelial cells (EC) using chemically defined media are provided, allowing EC production from early mesoderm progenitor cells in a short three days and EC production from pluripotent stem cells in a short five days. Media, isolated cell populations, and kits are also provided.

Provided is a method for inducing CD4−CD8+ T cells having an antigen specific cytotoxic activity from pluripotent stem cells, comprising the steps of: (1) differentiating pluripotent stem cells to give a cell culture comprising CD4−CD8− T cells and CD4+CD8+ T cells, (2) removing CD4−CD8− cells from the cell culture obtained in step (1), and (3) differentiating the CD4+CD8+ cells in the cell culture into CD4−CD8+ T cells.

A cell population comprising Corin- and / or Lrtm1-positive cells was produced by the following steps (1) and (2), from which Corin positive and / or Lrtm1 positive cells are collected using a substance that binds to Corin and / or a substance that binds to Lrtm1, and dopaminergic neuron progenitor cells are produced by performing suspension culture of the Corin positive and / or Lrtm1 positive cells in a culture solution containing one or more nutritional factors:(1) a step of performing adhesion culture of pluripotent stem cells in a medium for maintaining undifferentiated state containing a Sonic hedgehog (SHH) signal stimulant, and an undifferentiated state-maintaining factor in the absence of feeder cells but in the presence of an extracellular matrix, and(2) a step of culturing the cell population obtained in the step (1) in a culture solution containing one or more differentiation-inducing factors.

Naive pluripotent stem cells are produced at high efficiency. A method for producing naive pluripotent stem cells, wherein the method includes the following steps: (1) a step that cultures primed pluripotent stem cells in primed pluripotent stem cell culture medium; and (2) a step that cultures the cells obtained in step (1) in naïve pluripotent stem cell culture medium, including at least one of the following steps: (i) a step that introduces miRNA selected from miRNA belonging to the let-7 miRNA family and similar sequences thereto into the cells cultured in steps (1) or (2) or causes said miRNA to be expressed in said cells; or, a step that introduces an activity inhibitor of a let-7 miRNA family target protein into the cells cultured in steps (1) or (2) or causes said activity inhibitor to be expressed in said cells; and (ii) a step that introduces a bone morphogenetic protein into the cells cultured in steps (1) or (2), causes said bone morphogenetic protein to be expressed in said cells, or adds said bone morphogenetic protein to the medium of (1) or (2); or a step that adds a bone morphogenetic protein activator to the medium of steps (1) or (2).

Provided is a method for inducing muscle-derived differentiation of porcine pluripotent stem cells, which does not involve transgenosis and is free of serum addition in the whole course, and also provided is a method for preparing cell cultured meat (CM) based on the muscle-derived differentiation induction method.