48 results about "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.

The invention provides a recombinant lentivirus capable of indicating bovine spermatogonial stem cell differentiation, a spermatogonial stem cell line and application, and belongs to the technical field of cell biology. The invention provides a method for constructing a spermatogonial stem cell line for indicating bovine sperm differentiation on the basis of a lentivirus transduction technology, and a visual detection system for differentiating spermatogonial stem cells into spermatoblasts is constructed by regulating expression of mCherry protein by utilizing a PRM1 promoter. According to the method, the expression of PRM1 is indicated by fluorescent protein mCherry, so that the differentiation process of the bovine spermatogonial stem cells is indicated, and a good tool is provided for the induction of differentiation of the bovine spermatogonial stem cells to sperms, the research of a development mechanism and the optimization of a differentiation system of the bovine spermatogonial stem cells. The method is a new way for solving the neck clamping problem that good-variety bovine sperms are difficult to obtain in scientific research and production, can also be used for optimization of a spermatogonial stem cell differentiation system, sorting and purification of sperms, even downstream research of drug screening and the like, and has a good application prospect.

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 application discloses a method for stably producing exosomes of regenerative repair and anti-inflammatory molecules of sheep umbilical cord mesenchymal stem cells and application thereof in skin care products, and comprises the following steps: S1, using healthy sheep umbilical cord as raw material, obtaining primary sheep umbilical cord mesenchymal stem cells through aseptic treatment, Wharton's jelly extraction and tissue block adherent culture; S2, introducing an immortalization gene into the primary sheep umbilical cord mesenchymal stem cells through a lentivirus transduction technology, and obtaining an immortalized sheep umbilical cord mesenchymal stem cell line through resistance screening; S3, collecting supernatant after amplification culture of the immortalized sheep umbilical cord mesenchymal stem cell line, and purifying through multi-stage centrifugal impurity removal and PEG precipitation method to obtain stably produced exosomes containing regenerative repair and anti-inflammatory molecules. The exosomes containing regenerative repair and anti-inflammatory molecules provided by the application exhibit significant advantages in yield and quality, and have outstanding application value in the fields of medical beauty anti-aging and skin care products.

The invention provides an immortalized mesenchymal stem cell line capable of simultaneously expressing IL1-RN and COL17AD protein active segments as well as a preparation method and application of the immortalized mesenchymal stem cell line, and belongs to the technical field of genetic engineering. The method specifically comprises the following steps: (1) constructing a pMSCV-BMI-P2A-TERT virus; (2) infecting the mesenchymal stem cells, and performing passage to obtain immortalized mesenchymal stem cells; (3) constructing a pMSCV-IL1-RN-P2A-COL17AD virus (pMSCV-IL1-RN-P2A-COL17AD And (4) infecting the immortalized mesenchymal stem cells by using the pMSCV-IL1-RN-P2A-COL17AD virus obtained in the step (3), and carrying out passage, so as to obtain the immortalized mesenchymal stem cell line capable of simultaneously expressing IL1-RN and COL17AD protein active segments. The cell supernatant of the cell line provided by the invention has the function of promoting skin injury repair, and can be applied to functional cosmetics and skin repair drugs.

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 belongs to the technical field of cell culture, and particularly relates to a construction method of an immortalized goat umbilical cord mesenchymal stem cell line. The method comprises the following steps: by taking newborn goat lamb umbilical cord tissues as a research object, cleaning goat lamb umbilical cords, stripping blood vessels, separating Wharton's jelly, cutting the Wharton's jelly into 1mm < 3 > tissues, culturing by using a DMEM / F12 complete culture medium containing fetal calf serum, carrying out passage, cryopreservation, repeated resuspension and centrifugation, and inoculating into a culture dish for culture; lentiviruses containing SV40 large T antigens are packaged in vitro and are transduced into goat umbilical cord mesenchymal stem cells, and immortalization is realized after long-term subculture. The result of the embodiment shows that the construction method of the immortalized goat umbilical cord mesenchymal stem cell line provided by the invention is strong in repeatability, simple and efficient in operation steps, good in stem cell growth state and stable in physiological state.

The invention provides a trachinotus ovatus spermatogonial stem cell line as well as establishment, cryopreservation and recovery methods and application thereof, and relates to the technical field of biologication.The method comprises the steps that tissue cells of trachinotus ovatus testis are placed in a complete culture medium to be cultured, and a fresh culture medium is replaced once every 2-4 days within 15 days before culture; culturing until the bottom of the bottle is full of cells, adding a trypsin solution to digest the cells, sucking out the digested cells every 30 seconds, respectively adding a complete culture medium for culturing for three times, screening out high-proportion spermatogonial stem cells for subsequent continuous culture, subculturing to a new culture bottle according to the proportion of 1: (1-3), and culturing to obtain the trachinotus ovatus spermatogonial stem cell line. The trachinotus ovatus spermatogonial stem cells with relatively high purity are obtained by a differential digestion method, and the trachinotus ovatus spermatogonial stem cell line is obtained by setting a passage proportion for culture and can be stably passed. And gene transfection research is carried out, so that technical support is provided for extensive research on spermatogonial stem cells of trachinotus ovatus.

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.

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 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 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 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.