47 results about "Embryonic Stem Cell Line" patented technology

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 human-derived recessive retinal degeneration disease organ-like model based on Prom1 gene knockout. Specifically, the invention provides a specific gRNA targeting a Prom1 gene exon 9, and the specific gRNA can be used for efficiently knocking out the Prom1 gene of the human embryonic stem cell. The invention also provides a Prom1 gene knockout stem cell line derived from the human embryo pluripotent stem cell H9 and a retinal organ disease model. The retina-like organ can provide a research model for clinical treatment drugs.

The present invention discloses a method for establishing and applying a caudal fin-induced pluripotent stem cell line of Erythroculter ilishaeformis. By the tissue adherence method, a cell line with high proliferation performance and stemness is isolated from the caudal fin of Erythroculter ilishaeformis. In the early stage, the cells of this cell line are significantly fibroblast-like. Later, the cells become smaller but still maintain a spindle shape, without obvious contact inhibition phenomenon, and can spontaneously form cell clusters similar to induced pluripotent stem cells. Alkaline phosphatase staining is positive and it expresses stemness marker genes such as Oct4, Sox2, Klf4, Nanog, and c-Myc. The present invention uses a single cAMP agonist small molecule compound to reverse the phenomenon of weakened stemness after continuous subculture of this cell line, enabling it to maintain the characteristics of high proliferation performance and high stemness, which is suitable for future research on the induction and differentiation of fish stem cells and provides more choices for the research and application of fish stem cells.

This invention relates to the field of organoid disease models, and discloses a method for constructing a human brain organoid model of early-onset Alzheimer's disease (AD) and its applications. This invention introduces [a specific technology] into human embryonic stem cell lines through single-base editing and lead editing techniques. PSEN1 ΔE9、 PSEN1 M146V and APP Human embryonic stem cell lines carrying four familial pathogenic gene mutations (K670N and M671L) were constructed and induced to differentiate into Alzheimer's disease (AD) brain organoids. The AD brain organoid model established by this invention exhibited tau phosphorylation pathological phenotypes as early as 20 days and Aβ-related phenotypes as early as 40 days, with increased total Aβ, decreased Aβ42 / Aβ40 ratio, and simultaneous aggravation of Aβ-Tau pathology. This represents a complex neurodegenerative pathological model where multiple mutations synergistically regulate Aβ production and tau phosphorylation, which is highly valuable for understanding and studying early-onset familial AD. This invention provides a human brain organoid model for in vitro AD studies and offers a tool for studying the pathological mechanisms of AD and screening drugs.

This invention discloses a method for inducing and culturing burdock stem cells and stem cells, belonging to the field of plant biotechnology. Using burdock roots as explants, the cambium is determined by staining, and the cambium is obtained using enzymatic digestion. The cambium is then inoculated into a specific ratio of induction solid medium and cultured in the dark to efficiently induce primary stem cells. These primary stem cells are then transferred to a proliferation solid medium for subculture and selection to obtain and maintain a vigorous, soft stem cell line. The solid stem cell line is further transferred to a liquid medium for shaking culture to establish a suspension stem cell line. This invention provides high induction efficiency, stable stem cell characteristics, and good reproducibility, offering a high-quality starting material and technical platform for in vitro preservation of burdock germplasm, large-scale production of secondary metabolites, and genetic transformation.

This invention belongs to the medical field and provides a human trophoblastic stem cell line derived from complete hydatidiform mole and its applications. The human trophoblastic stem cell line, with accession number CGMCC No. C2024411, can be induced to differentiate into syncytiotrophoblast and extravillous trophoblast, mimicking the differentiation and developmental defects of the trophoblastic lineage in hydatidiform mole. It can be used as an in vitro research model for complete hydatidiform mole and has broad application prospects in drug target research and genetic studies.

The invention discloses a line establishment method and a culture system of bovine trophoblast stem cells. The stem cell line is obtained by separation and induction from bovine blastocyst. The embryonic stem cell line has stability. The method can be applied to various life science and medical fields such as bovine animal breeding and breeding, gene editing models, animal cloned donor cells, medical musculoskeletal injury experiments, drug development carriers, vaccine production cell banks, embryo quality analysis and the like, and can be applied to large-scale production and application.

The invention belongs to the technical field of gene engineering, and particularly relates to a neural crest stem cell tracing system based on P75NTR-T2A-eGFP and application. The invention focuses on construction and application of a tracing system for differentiating pluripotent stem cells into neural crest stem cells in vitro. By constructing a Crispr / Cas9 vector, a termination codon region of a P75NTR gene expression cassette is precisely targeted. Meanwhile, a gene knock-in vector containing an (LHA) left homologous arm, T2A-eGFP-right homologous arm (RHA) is constructed. The two vectors are introduced into pluripotent stem cells (hPSCs) by means of electric shock transfection. Through the operation, the P75NTR-T2A-eGFP gene knock-in pluripotent stem cell line is successfully obtained, a foundation is laid for building a follow-up tracing system, and the P75NTR-T2A-eGFP gene knock-in pluripotent stem cell line has important application value in the field of neural crest stem cells.

The invention belongs to the field of medicine, and provides a complete grape embryo-derived human trophoblast stem cell line and application thereof. The preservation number of the human trophoblast stem cell line is CGMCC No.C2024411, and the human trophoblast stem cell line can be induced and differentiated into a syncytial trophoblast and an extravillus trophoblast, can simulate differentiation and development defects of a grape embryo trophoblast pedigree, can be applied as an in-vitro research model of a complete grape embryo, and has a wide application prospect in drug target research and genetic research.

The invention provides a construction method of a CETN3 knockout cell line, a human brain organ with microhead deformity and application of the human brain organ, and belongs to the field of organoids. The CETN3-deficient stem cells prepared by the method provided by the invention still have normal multiplication capacity and differentiation potential, do not influence the development of organoid, can still develop into organoid with good morphology, but have reduced volume, which is similar to phenotypes appearing in small head malformation patients. The CETN3 knockout embryonic stem cell line is utilized, the cells can be self-organized into a structure similar to the human cerebral cortex through nerve induction and 3D culture, and the phenotype of a small head malformation patient is simulated. By utilizing the organ, the research on the disease mechanism can be promoted, and the medicine can be screened more effectively and conveniently.

The application discloses a kind of construction methods and applications of motor neuron MNX1 reporter cell lines, belong to gene editing and stem cell technical field.The application develops a kind of expression motor neuron MNX1 reporter gene vector system by screening high-activity sgRNA, and vector system includes the guide vector of single-stranded guide RNA targeted to the vicinity of the termination codon of human motor neuron MNX1 gene, and the donor vector carrying the homologous arm in the vicinity of the termination codon of human motor neuron MNX1 gene.The application also includes the construction method of MNX1 reporter gene human pluripotent stem cell line into the above-mentioned vector system, the application can be applied to identify stem cell differentiation source human motor neuron, and the morphology and function of motor neuron are researched under the condition of living cell, provide important platform for the pathogenesis research of muscle-related disease, treatment method evaluation and new treatment target exploration.

This invention relates to the field of bovine embryonic stem cell culture technology, and particularly to a long-term culture system for morphological bovine embryonic stem cells completely without a feeder layer and its applications. After trying various culture methods, this invention establishes a feeder-free bovine morphological embryonic stem cell line (FF-bESCs). Bovine morphological embryonic stem cells cultured under the feeder-free conditions provided by this invention can maintain stable proliferation capacity and pluripotency over a long period. The bovine morphological embryonic stem cells provided by this invention can undergo gene editing and can be effectively induced to differentiate into primordial germ cell-like cells (PGCLCs). This invention lays an important foundation for the standardized and large-scale culture of bovine embryonic stem cells under feeder-free conditions, and for their application in stem cell breeding and other fields.

The invention relates to a construction method of a human haploid blastoid, which comprises the following steps: firstly, sorting haploid cells, and placing the haploid cells in a human blastoid induction system to generate a human haploid blastoid-like structure with haploid attributes, namely the human haploid blastoid; the constructed human haploid blastoid has a structure and cell composition similar to those of a human blastocyst, and human embryonic stem cell lines, human trophoblast stem cell lines and human extraembryonic endoderm stem cell lines can also be established in different cell induction culture solutions. The human haploid blastoid has the structure and function similar to those of a human blastocyst, also has the advantages of haploid cells, and can be used for research on generation of human haploid embryo related cell lines, genetic screening of human early embryonic development and the like.

The invention discloses an Alzheimer's disease risk gene defect nerve cell as well as a preparation method and application thereof. The engineered induced pluripotent stem cell comprises an induced pluripotent stem cell which is deficient in ABCA7 gene and expresses a neural differentiation transcription factor in an inducible manner. According to the present invention, the Alzheimer's disease (AD) risk gene defect is introduced into the existing pluripotent stem cell line through the CRISPR and other gene editing means, and the inducible expression neural differentiation transcription factor is introduced through the lentiviral vector so as to establish the engineered AD risk gene defect pluripotent stem cell line, the engineered AD risky gene defect pluripotent stem cell line can be used for rapidly preparing nerve cells with Alzheimer's disease related A beta pathological phenotypes in a large scale by adding antibiotics for induced differentiation and screening culture, and is used for researching AD related pathogenesis and intervention means.

The present invention relates to a ground extracellular matrix (ECM) produced by a human stem cell line with chondrogenic activity, wherein the human stem cell line is genetically modified to overexpress Bone Morphogenetic Protein-2 (BMP-2). The invention further relates to a composition comprising said extracellular matrix, and a gel-like substrate or precursor thereof. The invention further relates to a kit comprising said composition. The invention further relates to a method of manufacturing said composition. The invention further relates to a method for bone and / or bone marrow formation in vivo. The invention further relates to a method for testing a candidate pharmaceutical product. The invention further relates to a method for identifying treatment responders. The invention further relates to said composition for use in medicine.

The invention discloses an establishment method of induced pluripotent stem cells capable of overcoming immunogenicity and derived mesenchymal stem cells of the induced pluripotent stem cells, belongs to the technical field of biology, and solves the problems of limited passage times, large individual difference and immunological rejection of the mesenchymal stem cells in the prior art. The method comprises the following steps: reprogramming starting somatic cells to establish an induced pluripotent stem cell line, and continuously picking monoclonal passage to obtain a monoclonal induced pluripotent stem cell line; and performing beta2-microglobulin knockout on the monoclonal induced pluripotent stem cell strain to obtain beta2-microglobulin knockout induced pluripotent stem cell monoclone, namely the induced pluripotent stem cell capable of overcoming immunogenicity. The invention develops a method for inducing wild type and B2M knockout type hiPSC to differentiate into the iMSC by regulating an embryonic development signal channel to generate the iMSC, and the differentiated iMSC-WT and iMSC-KO are extremely similar to UCMSC in phenotype.

The present invention relates to a fluorescently-labeled stem cell line with hepatic stellate cell activation markers, and a method for screening for liver fibrosis-inducing factors or fibrosis-inhibiting factors by using same. A human induced pluripotent stem cell line for screening for liver fibrosis-inducing factors or fibrosis-inhibiting factors, prepared according to the present invention, enables screening for an inducing substance capable of inducing liver fibrosis or a substance capable of inhibiting liver fibrosis in a state in which cells are alive in hepatic stellate cells derived from a pluripotent stem cell line, and thus can be used for imaging-based high-throughput screening (HTS) compound discovery and for discovering liver fibrosis-inducing substances or fibrosis-inhibiting substances by using high-content screening equipment.

The invention relates to the technical field of biology, and particularly provides a glycyrrhiza glabra stem cell line and application thereof. The glycyrrhiza glabra stem cell line is preserved in the China General Microbiological Culture Collection Center, the preservation number of the glycyrrhiza glabra stem cell line is CGMCC No.46600, the glycyrrhiza glabra stem cell line is named as glycyrrhiza glabra stem cell line gc001, and the glycyrrhiza glabra stem cell line has high secondary metabolite content and can be directly used for high-end skin care products and medical products, and the glycyrrhiza glabra resource value is expanded.

The present invention discloses a method for deriving Holstein bovine embryonic stem cells from vitrified blastocysts. The method combines frozen blastocysts with a culture medium specific for the embryonic stem cell derivation stage to establish a stable bovine embryonic stem cell derivation system, achieving an 80% or greater probability of blastocyst adhesion and outgrowth, and an embryonic stem cell line establishment efficiency of 50% or greater. This method addresses the time-consuming issue of deriving bovine embryonic stem cells from fresh embryos, improves the efficiency and stability of bovine embryonic stem cell line establishment, facilitates the efficient establishment of in vitro breeding models, accelerates the breeding process, and reduces breeding costs, possessing significant practical value for the development of new dairy cattle breeds (lines).

The invention belongs to the technical field of biological medicines, and discloses an induced pluripotent stem cell (iPSC) capable of retaining NFIA gene deletion mutation and application of the iPSC. The stem cell strain is preserved in the China Center for Type Culture Collection (the preservation number is CCTCC NO: C202590), carries NC000001.10: g.6165096761842967del heterozygous deletion mutation, contains deletion of exons 3-6 of an NFIA gene, stably expresses pluripotent markers (OCT4, SOX2, NANOG and SSEA4), and has the capability of differentiating to three germ layers. The preparation method comprises the following steps: separating PBMC (peripheral blood mononuclear cells) from peripheral blood of a patient, carrying out reprogramming by utilizing 2.0 Sendai Reprogam Kit, and amplifying pluripotent clone in Essental 8TMMedium. The stem cell strain can be used for constructing a research model of neurodevelopmental disorder diseases (such as corpus callosum insufficiency, hydrocephalus and development retardation), simulating pathological phenotypes by differentiating neurons, glial cells or brain organs, and is also suitable for drug screening and gene therapy research. And a precise humanized tool is provided for mechanism analysis and treatment development of NFIA deficiency related diseases.

The invention provides a method for constructing a vascularized liver organ and the vascularized liver organ, and belongs to the technical field of tissue engineering. The method for constructing the vascularized liver organoid comprises the steps that a plurality of vectors in a transposition vector plasmid kit are transfected into pluripotent stem cells to obtain a stem cell line capable of regulating and controlling ETV2 gene expression, and the transposition vector plasmid kit can regulate and control ETV2 gene expression through non-natural amino acid; adding non-natural amino acid and a liver induction culture medium, inducing stem cell line differentiation and regulating ETV2122 * full-length protein expression, so as to obtain the vascularized liver organ. According to the application, reading-through of ETV2 gene full-length protein is realized by utilizing a gene codon extension technology, conditional expression of a vascular development key factor ETV2 is accurately controlled, efficient vascularization of liver organs is realized, and a new platform is provided for in-vitro liver development research, disease simulation, drug screening, toxicity detection and the like.

The invention relates to a conditional gene knockout / knockin animal model construction method based on a Cre-ES cell system. Firstly, an embryonic stem cell line (Cre-ES) capable of stably expressing Cre recombinase is established from a blastocyst of a tissue-specific Cre tool mouse, the normal karyotype rate of the embryonic stem cell line is larger than or equal to 85%, Cre genes are stably integrated, the chimeric rate after blastocyst injection reaches 100%, and the birth rate is larger than 20%. By utilizing the cell line, a gene editing carrier (such as a loxP-Stop-loxP report system targeting ROSA26) containing a loxP site is introduced into a cell through electrotransfection, positive clones are obtained through G418 screening and then injected to an ICR receptor blastocyst, and an F0-generation gene editing animal is directly obtained. The genotype of the F0-generation animal is flox / flox, and the F0-generation animal carries Cre recombinase, so that tissue specific editing can be realized without mating with a tool mouse. According to the method, the traditional four-generation breeding period is shortened to one generation, the efficiency is improved by 70%, the cost is reduced by 50%, the problems of line mixing, Cre leakage expression and marker gene residue are solved, and the method is suitable for space-time specific gene function research and disease model construction.

The invention discloses a bovine ectoderm stem cell line as well as an establishment method and application thereof. The bovine ectoderm stem cell line has the pluripotency of a bovine intermediate state stem cell line, expresses one or more pluripotency markers, can be stably passaged, still maintains pluripotency transcriptome characteristics and normal karyotype similar to those of an intermediate state (Formative) ectoderm cell after more than 112 generations, and has the potential of being differentiated into three-germ layers. The bovine intermediate-state stem cell line has the potential for myogenic differentiation, primordial germ cell-like cell differentiation and serving as somatic cell nuclear transplantation (SCNT) donor cells, which shows that the bovine intermediate-state stem cell line has wide application prospects in the aspects of promoting cell culture meat production and animal breeding.

The invention provides a method for promoting induction of human totipotent stem cells, belongs to the technical field of biology, and particularly relates to a method for promoting induction of the human totipotent stem cells to the totipotent stem cells through OTX2 knockout or knockdown, the method comprises the following steps: S1, constructing an OTX2 knockout or knockout plasmid vector; s2, establishing a human pluripotent stem cell line with OTX2 knocked down or knocked out; and S3, inducing the human pluripotent stem cells of which the OTX2 is knocked down or knocked out into totipotent stem cells, which are also called 8-cell-like cells (8CLCs). According to the invention, the efficiency of inducing the human pluripotent stem cells into the totipotent stem cells can be improved, and the application of the totipotent stem cells in in-vitro blastocyst-like induction, in-vivo chimeric embryo formation and the like is provided.

The present invention relates to a novel stem cell line derived from a Paralichthys olivaceus fillet and use thereof and, more specifically, to: a stem cell line (Accession No. KCLRF-BP-00545) derived from a Paralichthys olivaceus fillet; cultured meat comprising the stem cell line; and a food composition comprising the cultured meat. The stem cell line according to the present invention has pluripotency, thus enabling differentiation into various types of cells, exhibits an excellent growth rate even at low FBS concentrations, and is easily stored, and thus can be effectively used as a main raw material for the mass production of economical aquatic cell-cultured foods.

The invention discloses a healthy individual-derived autologous tumor stem cell line establishment method, which is characterized by comprising the following two stages: a first stage, inducing healthy human-derived somatic cells into hiPSC; in the second stage, the hiPSC is induced to be converted into the tumor stem cells, specifically, firstly, a tumor-derived conditioned culture medium is prepared, then under the synergistic effect of the tumor-derived conditioned culture medium and an induction adjuvant, the hiPSC is induced to be converted into the tumor stem cells, and the healthy individual-derived tumor stem cells are obtained after culture. According to the invention, the somatic cell-derived iPSC of healthy people can be induced into tumor stem cells, and the method can be well applied to cancer prevention, drug screening and health management.

The invention provides a preparation method of a low-immunogenicity induced pluripotent stem cell line and a method for preparing low-immunogenicity hematopoietic stem / progenitor cells from the cell line through induced differentiation, efficient differentiation from B2M gene knockout iPSC to CD34 + CD45 + hematopoietic stem / progenitor cells is realized, and a new stem cell source is provided for clinical research and application.

This invention discloses a method for inducing blastocysts to form pluripotent embryonic stem cells, thereby constructing a new pluripotent embryonic stem cell line with highly efficient chimeric fetal capacity, which can completely replace the function of epiblast cells in blastocysts. This provides a new model for studying the epigenetic mechanism and transcriptional status of embryonic stem cell pluripotency, and has great application potential for studying mammalian development and generating mouse models.

Embodiments of the present disclosure include compositions and methods related to engineered monoclonal TCR armed genetically engineered T (TARGET) cells for ready-to-use use for clinical therapy. In particular embodiments, the TARGET cells are produced from a pluripotent stem cell line and thus can be infinitely supplied. The TARGET cell products can be conventional alpha beta T cells, gamma delta T cells and unconventional T cells such as constant natural killer T (iNKT) cells, non-constant natural killer T (NKT) cells, mucosa related constant T (MAIT) cells and the like. The TARGET cell product can also be designed as a CD8-positive or CD4-positive T cell subtype, and the CD4-positive TARGET cell can be further designed as a variety of TH polarized subtypes.

Disclosed herein is a method of isolating a stem cell line derived from a tissue from a primary meristem in a leaf of a plant, including the steps of sterilizing at least a part of the leaf containing the central or other vein; cutting through the central or other vein of the leaf to expose primary meristematic cells from the central or other vein inner vascular tissue; and culturing cells from said exposed cells, which circumvents the difficulties of surface sterilization when other types of explants are used. Also disclosed herein are stem cell lines obtained using the method.