1418results about "Germ cells" patented technology

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.

The invention discloses a high-temperature-resistant klebsiella pneumoniae phage, a composition of the high-temperature-resistant klebsiella pneumoniae phage and a kit, the high-temperature-resistant klebsiella pneumoniae phage is klebsiella pneumoniae phage P6AKPJS20, the preservation number of the high-temperature-resistant klebsiella pneumoniae phage is CCTCC (China Center For Type Culture Collection) NO: M20242965, and the high-temperature-resistant klebsiella pneumoniae phage has a nucleotide sequence as shown in SEQ ID No.1-6. The bacteriophage has broad-spectrum bactericidal ability, wide acid-base tolerance range and certain tolerance to chloroform and diethyl ether, and can still maintain high activity while splitting target bacteria; meanwhile, the bacteriophage has good tolerance to high temperature and can adapt to various application conditions in production.

The invention provides a method for constructing a chronic unpredictable negative stress model, the chronic unpredictable negative stress model and application of the chronic unpredictable negative stress model in mental disease research, and belongs to the field of brain organ in-vitro model construction. Through dual verification of a microelectrode array and single cell sequencing, an in-vitro brain organ model capable of simulating chronic unpredictable negative stress core characteristics is successfully constructed, and a potential neural network function reconstruction mechanism and a cell molecule basis of the in-vitro brain organ model are disclosed. The comprehensive research normal form breaks through the limitation of a traditional single technology platform, and an accurate and efficient innovative research platform highly related to human is provided for pathogenesis research of mental diseases, especially chronic stress related diseases such as depression and anxiety and development of novel treatment strategies.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetically modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.

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.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a Myo1B protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

The invention provides preparation, sorting and application of LGE / CGE organoid and GABAergic neurons, and belongs to the field of brain organoid differentiation and GABAergic neurons. The preparation method of the LGE / CGE type organ comprises the following steps: S1, performing embryoid formation and initial culture on the 0th-4th day; s2, performing nerve induction and matrigel embedding on the 5th-11th day; s3, culturing the brain organoid on the 12th day to the 19th day; s4, dynamically culturing and screening the brain organoid on the 20th-35th day; and S5, carrying out differentiation and collection on LCOs of the LGE / CGE type organs after the 35th day. According to the method, the LGE / CGE organs containing the high-purity GABA energy neurons are obtained through differentiation from the brain organs for the first time, the high-purity LGE type GABA energy neurons and the high-purity CGE type GABA energy neurons are obtained through a specific sorting method, and the method has the deep application potential in the fields of disease model construction, medicine research and development and the like.

The invention belongs to the technical field of medicines, and discloses application of exosomes derived from human pluripotent stem cells in preparation of anti-hepatic inflammation medicines. According to the invention, the exosome derived from the human pluripotent stem cells is applied to the preparation of the anti-hepatic inflammation medicine, especially the preparation of the anti-hepatic fibrosis anti-inflammatory medicine. The human embryonic stem cell-derived exosome can inhibit and regulate polarization of liver macrophages, inhibit inflammatory liver macrophages, up-regulate anti-inflammatory liver macrophages, reduce secretion of inflammatory factors, improve secretion of anti-inflammatory factors and inhibit liver inflammations and development thereof, has an obvious anti-inflammatory treatment effect on inflammation of liver fibrosis, and can be used for preparing medicines for treating liver fibrosis. Therefore, the progress of hepatic fibrosis can be inhibited.

The invention relates to the technical field of biology, in particular to a composition for preparing a brain organ, the brain organ and application of the brain organ. The invention further discloses a construction method of the brain organ, and on the basis of the method, different stem cells such as ESCs and iPSCs can be utilized to stably construct the three-dimensional brain organ model capable of simulating the medulla oblongata trigeminal nerve. The invention further discloses an organoid assembly which has important application potential in the fields of research on development, functions and related diseases of brain nuclei, drug discovery and the like.

The present technology relates to a systems for quantifying rheotaxis in a sperm-containing sample. The system includes a microfluidic system having an inlet for charging fluids into a passage. One or more probes defining a confinement region suitable for retaining motile sperm are within the passage. The system further comprises an image processing computing device for obtaining a sequence of images of the confinement region of at least one of the one or more probes having motile sperm retained therein. The sequence of images of the confinement region is processed to determine a signal intensity value for said sequence of images, wherein said signal intensity value is based on a concentration of the motile sperm located in the confinement region at the flow rate. A rheotaxis quality value is determined for said sperm-containing sample based on the signal intensity value. Methods for quantifying rheotaxis in a sperm-containing sample are also disclosed.

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and / or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

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.

The invention discloses a double-gene screening expression vector of a CHO monoclonal cell strain and a preparation method and application of the double-gene screening expression vector, the double-gene screening expression vector comprises a glutamyl synthetase GS screening gene and an antibiotic screening gene, the antibiotic screening gene is subjected to weakening expression treatment, that is, an IRES sequence exists in front of the antibiotic screening gene, and an IRES sequence exists in front of the antibiotic screening gene. A target expression gene and an antibiotic screening marker gene share one set of promoter and terminator, and when the double-gene screening expression vector disclosed by the invention is used for screening CHO monoclonal cell strains, the screening efficiency can be remarkably improved, the screening time is saved, and the target gene has high expression quantity.

The invention provides a method for overspeed preparation of gene edited goldfish. The method comprises the following steps: S1, obtaining gobiocypris rarus without primordial germ cells as receptor fish; s2, designing a target spot for the goldfish amh gene, introducing a Cas9 / gRNA complex into a goldfish fertilized egg, performing sequencing verification to obtain a mutant individual, and feeding the mutant individual as a donor fish; s3, taking gonads of donor fish, cutting into pieces, performing enzymolysis digestion, filtering, and performing density gradient centrifugation to enrich donor germline stem cells; s4, transplanting the germline stem cells of the donor fish into the receptor fish body, and feeding to obtain the gene edited goldfish sperms and ova. S5, hybridizing and feeding goldfish amh mutant sperms and ova produced by gobiocypris rarus to obtain gene edited goldfish individuals. According to the invention, gobiocypris gobiocypris is used as a receptor, the cross-subfamily gene edited goldfish is used as a donor, and an efficient, stable and easy-to-popularize method for preparing the gene edited goldfish is provided by utilizing a belly borrowing reproduction technology.

Disclosed herein are non-human animals (e.g., rodents, e.g., mice or rats) genetically engineered to express a human or humanized T cell receptor (TCR) from a human or humanized TCR locus comprising a non-human TCR non-coding sequence, and optionally a humanized T cell co-receptor (e.g., humanized CD4 and / or CD8 (e.g., CD8α and / or CD8β)), and / or a human or humanized major histocompatibility complex that binds the humanized T cell co-receptor (e.g., human or humanized MHC II (e.g., MHC II α and / or MHC II β chains) and / or MHC I (e.g., MHC Iα) respectively, and optionally human or humanized β2 microglobulin). Also provided are embryos, tissues, and cells expressing the same. Methods for making the genetically engineered animals are also provided. Methods for using the genetically engineered animals for developing human therapeutics are also provided.

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and / or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

The invention discloses a zebra fish spermatogonial stem cell separation and transplantation method, and belongs to the technical field of fish germline stem cell separation. The method comprises the following steps: washing complete sexual glands of zebra fish testis by adopting penicillin / streptomycin double-antibody-containing PBS (Phosphate Buffer Solution), physically shearing, digesting and dissociating, and filtering by a screen, so as to obtain zebra fish testis cell suspension; performing density gradient centrifugation to obtain zebra fish spermatogonial stem cells; dyeing, marking, centrifuging, washing, re-suspending, and carrying out micro-injection transplantation to enter the back of the abdominal cavity of the acceptor zebrafish juvenile fish. According to different physical properties of different cells, spermatogonial stem cells are separated, purified and transplanted from zebra fish testis cell suspension by adopting a density gradient centrifugation method, and migration and proliferation processes of donor germ cells can be observed in receptor fish after fluorescence labeling. And a new technical path is provided for fish germplasm resource preservation, endangered fish protection, economic cultured fish yield increase and new variety cultivation.

The invention discloses a method for separating, purifying and transplanting spermatogonial stem cells of lateolabrax japonicus, the spermatogonial stem cells of lateolabrax japonicus are separated and purified, marine fish larvae or freshwater fish larvae are used as receptor larvae, and then the spermatogonial stem cells of lateolabrax japonicus are transplanted to abdominal cavities of the receptor larvae. The spermatogonial stem cells obtained through separation are complete in number, structure and function and low in cell clustering rate. After the transplanted spermatogonial stem cells are subjected to fluorescence labeling, the migration and proliferation processes of donor germ cells can be observed in receptor fishes of marine fish fries or freshwater fish fries, so that the receptor fishes with the chimeric germline are obtained. According to the method, through xenotransplantation of the spermatogonial stem cells, a'belly borrowing reproduction 'breeding mode is provided for the marine fishes, a new way for gamete preservation and rapid propagation is provided for the marine fishes with the long reproductive cycle, and a new method and a new thought are provided for genetic improvement and germplasm resource preservation.

The invention discloses a pharmaceutical composition for delaying oocyte senescence and application thereof, and relates to the technical field of biological medicine, and the pharmaceutical composition comprises alpha-ketoglutaric acid, a bicarbonate buffer solution, an energy metabolism substrate, polyvinyl alcohol, hormone, a growth factor and insulin-transferrin-selenium. The pharmaceutical composition for delaying oocyte aging disclosed by the invention is beneficial to improving the maturation rate of oocytes, reducing the cytoplasm fragmentation rate and the cleavage fragmentation rate, and improving the cleavage rate and the blastocyst rate; according to the invention, the content of beta-galactosidase is reduced, the function of mitochondria in the aging process after ovulation of the mature oocyte is improved, the membrane potential of the mitochondria in the aging process after ovulation of the mature oocyte is stabilized, the activity of the mitochondria is improved, and the normalization of RNA methylation modification in the aging process after ovulation of the mature oocyte is promoted.

The invention discloses a chicken sex identification method based on fluorescence labeling, and belongs to the technical field of gene engineering. The chicken sex identification method comprises the following steps: co-transfecting a CRISPR / Cas9 system and a WPG-EGFP donor plasmid into chicken primordial germ cells PGCs, screening to obtain EGFP positive PGCs, micro-injecting the EGFP positive PGCs into a receptor chicken embryo, hatching to obtain an EGFP reproductive chimera chicken, and identifying the sex through fluorescence labeling, the CRISPR / Cas9 system comprises sgRNA of a targeted chicken W chromosome WPG gene; the WPG-EGFP donor plasmid is constructed by inserting an EGFP gene into a WPG gene locus through a homologous recombination strategy. According to the method, the sex of the chicken can be visually and rapidly identified. The method solves the problems that a traditional sex identification method is complex in operation and depends on sample ages, has the advantages of high efficiency and accuracy, and is suitable for sex control and screening of poultry such as laying hens and broilers.

The present invention generally relates to compositions and methods useful for treating a brain injury such as stroke, optic neuropathy, traumatic brain injury, and cerebral palsy. The methods include administering HMCs obtained by in vitro differentiation of pluripotent stem cells and / or extracellular vesicles (EVs) derived from such HMCs (HMC-EVs) into a subject.

The invention discloses a method for separating and transplanting spermatogonial stem cells of Chinese sturgeons, and belongs to the technical field of separation of germline stem cells of fishes. The separation method comprises the following steps: washing Chinese sturgeon testis tissues with an L-15 culture medium containing penicillin / streptomycin double antibodies, and carrying out physical shearing, digestive enzyme enzymolysis and screen filtration to obtain a Chinese sturgeon testis cell suspension; performing percoll density gradient centrifugation to obtain high-purity spermatogonial stem cells of the Chinese sturgeons; and dyeing, washing, resuspending and carrying out micro-injection transplantation to enter the gonad primordium reproductive ridge part of the juvenile acceptor fish. According to the physical characteristics of the spermatogonial stem cells of the Chinese sturgeons, the spermatogonial stem cells with high purity are obtained by adopting percoll density gradient centrifugation, and the purpose of abdomen borrowing reproduction is achieved by adopting a microinjection mode, so that the preservation of germ cells of endangered fishes and the reconstruction of artificial populations are realized; and a new technical approach is provided for protecting genetic resources of the Chinese sturgeons and breaking through artificial propagation bottlenecks.

The invention provides a culture medium composition and a method for preparing mesenchymal stem cells. The culture medium combination comprises a culture medium A, a culture medium B and a differential culture medium, the culture medium A comprises a basic culture medium and a first small molecule compound combination, the culture medium B comprises a basic culture medium and a second small molecule compound combination, and the differential culture medium comprises a basic culture medium and a third small molecule compound combination. According to the culture medium and the method, the MSC can be prepared through ES or iPSC, and the culture medium and the method can be widely applied to the fields of regenerative medicine, tissue engineering, immunotherapy and the like.