502 results about "Multipotential stem cell" patented technology

The invention relates to a heart organoid and an in-vitro preparation method thereof, and belongs to the technical field of stem cells. The invention provides a heart organoid. The heart organoid is obtained by induced culture of an adherent pluripotent stem cell mass; the adherent pluripotent stem cell mass is obtained by adherent culture of a suspended pluripotent stem cell mass; the suspension pluripotent stem cell mass is obtained by suspension culture of pluripotent stem cells. According to the in-vitro preparation process of the heart organoid, after pluripotent stem cells form balls, the pluripotent stem cell balls are subjected to staged induced differentiation under adherent culture, so that cell subtypes such as epicardial cells and the like can be generated without additional addition or additional stimulation of signal channels, and the heart organoid is more homogeneous. The self-occurring multi-lineage coexisting heart organoid can better simulate biological events derived from multiple cell types and multiple cell subtypes in the early development process of an embryonic heart, and has great application prospects in exploration of pathogenic factors and potential treatment targets of congenital heart diseases.

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.

The invention relates to the technical field of image processing, in particular to an induced pluripotent stem cell density detection method, computer equipment and a medium, and the method comprises the following steps: acquiring an induced pluripotent stem cell image; presetting a local area of each pixel point; obtaining a local brightness performance value according to the gray level distribution of the pixel points in the local area; obtaining local boundary ambiguity according to the gradient distribution characteristics; obtaining local form regularity according to the uniformity characteristic of gray level distribution in a pixel point local area; obtaining a density comprehensive performance value of each pixel point; clustering the pixel points to obtain a preset number of clusters; and respectively enhancing each cluster by adopting histogram equalization to obtain an enhanced induced pluripotent stem cell image, and carrying out density detection on the enhanced induced pluripotent stem cell image. The invention aims to enhance the induced pluripotent stem cell image and improve the accuracy of the cell density detection result.

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.

The invention provides construction and application of a human pluripotent stem cell-derived heart organoid traditional Chinese medicine extract toxicity test chip. The chip comprises a plurality of micro-cell culture chambers and a micro-fluidic pore channel, heart organoid culture is carried out in the micro-cell culture room, culture media are added in the micro-fluidic channel to maintain the activity of the heart organoid, and traditional Chinese medicine extracts are added to evaluate the toxicity of traditional Chinese medicine. By simulating the reaction of the heart organ, the toxicity of the traditional Chinese medicine can be evaluated, the safety and effectiveness information about the traditional Chinese medicine can be provided, finally, the use of experimental animals can be reduced, and the animal welfare can be improved.

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.

In various aspects, the invention provides culture platforms, cell culture media, and methods of differentiating pluripotent cells into hematopoietic cells. In certain aspects, the invention also provides pluripotent stem cell-derived hematopoietic cells generated using the media and methods disclosed herein. The pluripotent stem cell-derived permanent HE cells produced by the methods provided herein are capable of differentiating into hematopoietic lineage cells comprising T cell progenitor cells and T cells in addition to NK cell progenitor cells, NK cells, NKT cells, or B cells.

The present invention relates to a modified pluripotent cell and to a method for forward programming said cell. The invention further relates to use of said modified pluripotent stem cell for tissue engineering and to a food product comprising said modified pluripotent stem cell.

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.

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.

Disclosed is a method for producing T cells, the method comprising: (1) culturing three-dimensional cell aggregate(s) comprising cells that can differentiate into T cells, and stromal cells that express Notch ligand(s) derived from pluripotent stem cells. Also disclosed are T cells obtained by the method, and a medicine comprising the T cells.

The invention relates to an N2B27 culture medium for inducing generation of bidirectional pluripotent stem cells, one of LY2090314, AS1842856 or CHIR99021 is added into an N2B27 basal culture medium, and the concentration of the CHIR99021 is 10 mu M. The invention also relates to a preparation method of the N2B27 culture medium. The weight of the LY2090314 is 10 nM, and the weight of the AS1842856 is 0.6 [mu] M. The invention also provides an application of the culture medium in induced production of bidirectional pluripotent stem cells. According to the invention, a novel stem cell culture condition for promoting co-expression of genes OCT4 and CDX2 by adding small molecules LY2090314 (LY), AS1842856 (AS) and CHIR99021 to inhibit signal channels of FOXO1 and GSK3 is screened out. The culture conditions are simple and convenient, the application range is wide, the cultured cells keep the characteristics of 16-32 cell stages of the embryos, and the development characteristics of the embryos can be efficiently reproduced. Under the condition of not depending on transcription factors, the screened bidirectional pluripotent stem cells (BPSCs) can promote efficient generation of TSC cells and establish a TSC cell line through independent differentiation of a serum culture medium or induced differentiation of a TSC culture medium. The obstacles between early embryo pedigree are broken, and the research on early embryo development is promoted.

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 provides a preparation method and a kit for reprogramming induced pluripotent stem cells of PBMC (peripheral blood mononuclear cells), particularly provides PBMC reprogramming based on mRNA-LNP (peripheral blood mononuclear cells), and more particularly relates to a preparation method for reprogramming induced pluripotent stem cells of peripheral blood mononuclear cells. The preparation method comprises the following steps: preparing a nano liposome mRNA (messenger ribonucleic acid) cocktail-LNP; according to the method, the nano-liposome mRNA-LNP is used for inducing peripheral blood mononuclear cells to reprogram, so that reprogrammed induced pluripotent stem cells are obtained, and an iPSCs reprogramming technology which is low in cost, low in toxicity, efficient, stable, repeatable and safer is realized. The invention also provides a reprogramming kit and a preparation method of the induced pluripotent stem cells.

Methods for generating human vascular leptomeningeal cells (VLMCs) from human oligodendrocyte progenitor cells (OPCs) are provided using chemically-defined culture media that allow for generation of differentiated cells in a 34-day culture protocol. Methods of generating human VLMCs from human pluripotent stem cells using chemically-defined culture media in a 40 day culture protocol are also provided. Culture media, isolated cell populations, and kits are also provided.

The present invention provides methods of producing in vitro derived neutrophils or macrophages in xenogen- and serum-free conditions from pluripotent stem cells and in vitro derived populations of neutrophils and macrophages. Methods of treatment using in vitro derived neutrophils or macrophages are also contemplated.

The invention provides cell culture conditions for culturing stem cells, including feeder-free conditions for generating and culturing human induced pluripotent stem cells (iPSCs). More particularly, the invention provides a culture platform that allows long-term culture of pluripotent cells in a feeder-free environment; reprogramming of cells in a feeder-free environment; single-cell dissociation of pluripotent cells; cell sorting of pluripotent cells; maintenance of an undifferentiated status; improved efficiency of reprogramming; and generation of a naïve pluripotent cell.

Disclosed are: a method for producing a cell population containing regulatory T cells, the method comprising (1) culturing a cell population containing pluripotent stem cell-derived CD4+ T cells in the presence of at least one substance selected from the group consisting of a CDK8 and / or CDK19 inhibitor, a TNFR2 agonist, an mTOR inhibitor, and a TGF-βR agonist; a cell population containing regulatory T cells obtained by the method; and a medicine containing the cell population containing regulatory T cells.

The present invention provides a method for producing a cartilage-like tissue from pluripotent stem cells, the method comprising a step for culturing pluripotent stem cells in a cartilage differentiation culture medium and a step for increasing the ascorbic acid concentration of the cartilage differentiation culture medium. The present invention also provides a method for regulating the hardness of induced cartilage-like tissue differentiated from pluripotent stem cells, the method comprising a step for culturing pluripotent stem cells in a cartilage differentiation culture medium and a step for increasing the ascorbic acid concentration of the cartilage differentiation culture medium.

The present invention relates to a method for preparing organ-specific mesenchymal cells from pluripotent stem cell-derived intestinal organoid stromal cells. By using cells derived from stromal cell layers adjacent to intestinal organoids for differentiation into organ-specific mesenchymal cells, the present invention can greatly increase the efficiency of differentiation into stromal cells through the regulation of retinoic acid (RA) and hedgehog (HH) signaling pathways, and can increase the expression of organ-specific markers without exhibiting undifferentiated state cell characteristics, and thus mesenchymal cells having well-simulated biological characteristics can be prepared.

Described herein is a microphysiological system for models of disease. Specifically, induced pluripotent stem cells (iPSCs) and iPSC-derived cells, including those obtained from disease patients, are seeded onto microfluidic “chip” devices to study cellular development and disease pathogenesis. Herein, neurodegenerative disease modeling, including Parkinson's Disease (PD) is shown to reproduce key PD pathology in a vascularized human model that contains neurons relating to PD pathology. Such compositions and methods are used for research for PD biomarkers, patient screening for PD risk assessment, and therapeutic discovery and testing. A panel of biomarkers are generated through analysis of living PD-chips by neural activity, whole transcriptomic, proteomic, and metabolomic analysis, and functional enzyme tests of media and tissue. Introducing therapeutics through a vasculature channel, coupled with blood brain barrier penetration studies can be assessed for efficacy in the human neural cells present in the PD-Chip.

The invention provides a method for inducing human pluripotent stem cells to be differentiated into retinal pigment epithelial cells and application. The method comprises the following steps: performing adherent culture on pluripotent stem cells to serve as differentiation starting cells; the differentiation starting cells are cultured in stages with different culture media to obtain the retinal pigment epithelial cells, and the culture media in at least one stage comprise a WNT non-classical pathway activator. The invention further provides a composition and a culture medium for inducing the human pluripotent stem cells to be differentiated into the retinal pigment epithelial cells, the differentiation efficiency of the RPE can be remarkably improved, the cell harvesting purity is improved, and the composition and the culture medium are suitable for large-scale industrial production.

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 methot 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 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.

Provided herein are genetically-modified cells such as pluripotent stem cells and mesenchymal stem cells and methods for producing the same. The gene editing strategy of the present disclosure achieves high expression of a transgene in the genetically modified cells.

The invention relates to the technical field of biology, in particular to an expression vector structure and a method for generating multilineage hematopoietic stem / progenitor cells based on an expression vector. The cDNA sequences of hnCD16 and mbIL-15 are expressed in the same vector in series and integrated into a mammal pluripotent stem cell genome, the pluripotent stem cell capable of stably expressing hnCD16 and mbIL-15 can be obtained, the method is easy and convenient to operate and high in efficiency, hematopoietic stem / progenitor cells capable of expressing mbIL-15-hnCD16 can be obtained through induced differentiation, and the hematopoietic stem / progenitor cells can be used for preparing the hematopoietic stem / progenitor cells. And the mbIL-15-hnCD16-NK cell obtained by further induced differentiation has higher killing activity and longer survival time.

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.

Disclosed herein are methods, compositions, and culture media for generating synthetic embryos in vitro from mammalian pluripotent stem cells, such as pluripotent embryonic stem cells. In some embodiments, the methods can include co-culturing wild-type mammalian pluripotent stem cells with modified mammalian pluripotent stem cells that include one or more genes encoding transcription factors that can drive the generation of extraembryonic or extraembryonic-like cells (e.g., GATA6, SOX17, GATA3, and / or TFAP2C genes) in a culture medium under conditions that allow the pluripotent stem cells to self-organize into post-implantation embryonic structures. In some embodiments, the pluripotent embryonic stem cells are human pluripotent embryonic stem cells, and the generated synthetic embryo is a human embryo.