57 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 belongs to the technical field of stem cell exosome preparation, and provides a preparation method and therapeutic application of a stem cell exosome loaded with therapeutic molecules, and the preparation method comprises the following steps: S1, selecting and culturing a stem cell line with exosome secretion capability; s2, in the stem cell culture process, adding selected therapeutic molecules, so that the therapeutic molecules can be taken by the stem cells and integrated into exosomes of the stem cells; s3, collecting stem cell culture supernate, and separating and purifying the exosome through a differential centrifugation, ultrafiltration or density gradient centrifugation method; s4, identifying the exosome obtained by separation; a stem cell line with efficient exosome secretion capacity is selected and cultured, and selected therapeutic molecules are added in the culture process, so that efficient loading of the therapeutic molecules is realized; according to the method, the operation steps are simplified, the loading efficiency is improved, it is ensured that therapeutic molecules can be stably integrated into the stem cell exosome, and meanwhile the biological activity and stability of the exosome are kept.

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.

Homogeneous human amniotic fluid stem cell clones are provided. The clones serve as direct bioassays to test the effect of drugs, nutraceuticals, vitamins and toxic agents on fetus growth, differentiation and development, and may be administered in form of pharmaceutical compositions for reconstructive engineering and for the treatment of cardiac, neurological and osteoarthritic diseases.

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 relates to the technical field of biology, and discloses a novel non-integrated bovine induced pluripotent stem cell line construction method, which comprises the following steps: selecting a proper cell source, constructing an episomal vector system, introducing an episomal vector into bovine primary fibroblasts by using an electrotransfection method, and screening and culturing the transfected bovine primary fibroblasts in a culture medium to obtain the novel non-integrated bovine induced pluripotent stem cell line. According to the method, positive cells successfully transferred into dry factors are obtained, the cell morphology is monitored in the reprogramming process, along with the progress of reprogramming, more and more pluripotent clones appear in a cell population, amplification and identification are performed through clone culture, and the pluripotency and differentiation potential of the successfully established bovine induced pluripotent stem cells are verified. A stable bovine induced pluripotent stem cell line is established, long-term culture is carried out, and pluripotency and stability of the bovine induced pluripotent stem cell line are verified. Through an episomal non-integration method, the cattle non-integration induced pluripotent stem cell line which can be passaged for multiple times in vitro, is relatively good in hereditary stability and can maintain pluripotency and in-vitro differentiation capacity is constructed.

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 present disclosure relates to a real-time chemical screening method using hepatic organoids fluorescently labeled with cytochrome P450 1A1 enzyme. The method for screening an AHR modulator or a hazardous chemical using CYP1A1 fluorescently labelled human pluripotent stem cell line-derived hepatic organoids according to the present disclosure enables the detection of substances that induce CYP1A1 regulation with higher sensitivity than when using the conventional human pluripotent stem cell line-derived hepatocytes by using hepatic organoids that can mimic the human body, and thus can be usefully utilized for early screening of new toxic or carcinogenic compounds while replacing animal testing.

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 invention provides a serum-free culture method for converting hUC-MSCs into hiPSCs through induction of a small molecule compound, can be used for preparing pluripotent stem cells, and belongs to the field of biomedicine. Comprising thymine analogs, cAMP activators, TGF-beta receptor inhibitors, bone morphogenetic proteins, RA receptor activators, GSK3 inhibitors and basic fibroblast growth factors, and the culture system does not contain serum. And 2, in the process of carrying out micromolecule compound reprogramming on the hUC-MSCs by utilizing the method provided by the invention, frequent digestion, inoculation and tray separation on the hUC-MSCs are not needed, so that compared with the existing culture method, the operation steps are simplified, the cell loss generated in the digestion, inoculation and tray separation process is reduced, and the cell quality is improved. According to the invention, the reprogramming efficiency of the small molecule compound on hUC-MSCs is obviously improved; and 3, by detecting the co-expression (such as OCT4, SOX2 and NANOG) of transcription factors in reprogrammed cells of a small molecule compound reprogrammed pluripotent stem cell line obtained by the technology, it is proved that the hiPSCs obtained by inducing the hUC-MSCs have stable pluripotency capable of continuous passage.

Homogeneous human amniotic fluid stem cell clones are provided. The clones serve as direct bioassays to test the effect of drugs, nutraceuticals, vitamins and toxic agents on fetus growth, differentiation and development, and may be administered in form of pharmaceutical compositions for reconstructive engineering and for the treatment of cardiac, neurological and osteoarthritic diseases.

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 discloses application of NSUN2 in promoting self-renewal and line establishment of embryonic stem cells. The research finds that the NSUN2 can protect the self-renewal of the embryonic stem cells and inhibit the differentiation of the embryonic stem cells; the deletion of the NSUN2 gene has a remarkable adverse effect on the establishment process of an embryonic stem cell line, and the line establishment efficiency is greatly reduced, so that the NSUN2 gene plays a key role in self-renewal and differentiation regulation of embryonic stem cells, and the deletion of the NSUN2 gene can cause the establishment efficiency of the embryonic stem cell line to be remarkably reduced and influence the normal functions of the embryonic stem cells. Therefore, a new research direction and application prospect can be provided for regenerative medicine and disease treatment.