2618results about "Germ cells" patented technology

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

The invention 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.

The invention discloses a pig chimeric embryo culture medium and application thereof in effectively improving the in-vitro chimeric ability of pig embryonic stem cells, and belongs to the technical field of bioengineering. According to the pig chimeric embryo culture medium disclosed by the invention, a neutral culture environment for promoting synchronous development of stem cells and embryos is created by accurately regulating and controlling growth factors, nutrient substances and other key components in the culture medium. On the basis, overexpression of anti-apoptosis factors, optimization of microinjection conditions and adoption of a laser-assisted incubation means are combined, so that the chimeric efficiency is further improved, integration of the stem cells and embryos is effectively supported, the chimeric efficiency is improved, and a reliable in-vitro tool is provided for evaluating the chimeric ability and pluripotency of the pig stem cells.

The invention provides a porcine oocyte in-vitro maturation culture solution additive and application thereof, and belongs to the field of embryo engineering, and the additive contains taurine. The additive also contains sodium pyruvate, cysteine, glucose, NaHCO3, fetal calf serum, estradiol, pregnant horse serum gonadotropin and human chorionic gonadotropin. When the porcine oocyte in-vitro maturation culture solution added with the additive is used for culturing porcine oocytes in vitro, the ROS content in the porcine oocytes can be remarkably reduced, the oxidation resistance of the porcine oocytes can be improved, and the mitochondrial distribution of the oocytes can be improved, so that the development of the oocytes can be improved; the maturation of porcine oocytes and the development of early embryos are promoted. When the technology is applied to the pig assisted reproduction technology, the success rate and quality of embryo engineering can be improved; the pig oocyte in-vitro maturation culture solution added with the additive can improve the premature embryo division rate in parthenogenetic activation of the pig oocytes.

The invention discloses a sebastes schlegeli testis cell line as well as a construction method and application thereof, and belongs to the technical field of marine fish cell culture. Primary culture and subculture are carried out on sebastes schlegeli juvenile fish testis tissues, so that the sebastes schlegeli testis cell line capable of being continuously subcultured in vitro for a long time is established. The sebastes schlegeli testis cell line established by the invention is in a fibroblast shape, has a good growth curve, and has relatively strong multiplication capacity. The lentiviral vector can significantly express the stromal cell marker gene temp2, can transfect the cell line, and can be subsequently applied to research of gonad development related functional genes. At present, the cell line has been subjected to in-vitro continuous passage for more than 60 generations, can be subjected to cryopreservation, and is high in recovery survival rate. Establishment of the cell line provides a stable and excellent experimental material for research of fish gonad development genes.

Methods, compositions and devices characterized by in vitro maturation of oocytes. In particular, the present disclosure features methods of inducing oocyte maturation in vitro by co-culturing oocytes of a female subject with an ex vivo composition comprising a plurality of ovarian supporting cells (e.g., granular cells). Additional methods of administering a follicle triggering agent and extracting oocytes from a female subject are also provided. Such methods, compositions, and devices are particularly useful for assisted reproductive technology (ART) procedures.

The invention belongs to the field of stem cells, and particularly relates to a method for inducing porcine embryonic stem cells to differentiate skeletal muscle cells by using a complete small molecule compound. The method comprises the following steps: S1, an induction stage: culturing the porcine embryonic stem cells in a culture medium for two days, and replacing with culture medium culture in the induction stage; s2, a differentiation stage: a differentiation stage culture medium is used for culture. The method is simple in operation process and high in efficiency, and large-scale production of the cell meat is facilitated while differentiation of the porcine embryonic stem cells into the skeletal muscle cells is achieved.

The invention relates to a method for induced differentiation of skeletal muscle cells by porcine embryonic stem cells. Comprising the following steps: (1) constructing a PAX7-overexpressed porcine embryonic stem cell stable cell line; (2) induced differentiation of skeletal muscle cells; (3) performing immunofluorescence identification; wherein the step (1) specifically comprises the following steps: 1.1, constructing a DOX (deoxyribonucleic acid) induced PAX7 expression plasmid PiggyBac-TRE3G-EF1a-PAX7; 1.2, performing electrical transformation on the porcine embryonic stem cells cultured for two days, and adding Buffer E into an electrical transformation cup; taking Buffer R, mixing plasmid and PBase, uniformly mixing, and re-suspending cell precipitate; 1.3, after electrotransformation is finished, the cells are planted on the feeding layer, Y-27632 is added into the culture medium, and the culture medium without Y-27632 is replaced with the culture medium without Y-27632 on the second day; and 1.4, after culturing for 3-4 days, selecting the porcine embryonic stem cells which are successfully subjected to electrotransfection for amplification culture, and naming PAX7-PESCs.

The invention discloses for the first time pluripotent cells, including induced pluripotent stem cells, embryonic stem cells, hypo-immune pluripotent cells, cells that have been derived therefrom, and cells that have been biologically differentiated therefrom into particular tissue lineages that are ABO blood type O Rhesus Factor negative or otherwise evade rejection resulting from blood type antigen mismatch. The invention further provides universally acceptable “off-the-shelf” pluripotent cells and derivatives thereof for generating or regenerating specific tissues and organs.

The invention discloses a micro-fluidic chip for vitrification cryopreservation of oocytes or embryos and a preparation method and application of the micro-fluidic chip. The micro-fluidic chip comprises a micro-fluid channel layer, and the micro-fluid channel layer is arranged in the micro-fluid flowing direction; the microfluid channel layer comprises one or more solution inflow channels, a snakelike mixing channel, one or more shunt channels, a snakelike channel, a cell storage chamber and a waste liquid discharge channel which are communicated in sequence; wherein a first micro-column array is arranged at an outlet of the cell storage chamber, and a second micro-column array is arranged in a channel outside an inlet of the cell storage chamber; and a cell channel is communicated between the cell storage chamber and the second micro-column array. The micro-fluidic chip can realize the integration of operations such as vitrification cryopreservation and rewarming, reduces the cell transfer frequency, avoids pollution, improves the treatment efficiency, can significantly improve the survival rate of oocytes or embryos after rewarming, and can further promote the development of an assisted reproduction technology.

The invention relates to an application of a regulator in improving the maturation quality of frozen pig oocytes, the regulator is N-(2-(2-(4-chlorphenyl) thiazole-4-yl) ethyl) butyryl, and the regulator improves the maturation quality of the frozen pig oocytes by improving the mitochondrial function of the frozen pig oocytes after in-vitro maturation. The invention provides a new application of N-(2-(2-(4-chlorphenyl) thiazole-4-yl) ethyl) butyramide, namely the N-(2-(2-(4-chlorphenyl) thiazole-4-yl) ethyl) butyramide is used for developing a novel oocyte vitrification freezing solution and an in-vitro maturation culture solution, and the obtained high-quality oocytes can be beneficial to embryo biotechnology researches such as in-vitro fertilization and somatic cell nuclear transplantation.

The invention relates to the technical field of cell differentiation and the technical field of myocardial tissue regenerative medicine, in particular to a method for rapidly inducing directional differentiation of embryonic stem cells into myocardial cells in vitro. A matrix sandwich method is adopted, specific small molecule compounds are added in different time periods to induce the embryonic stem cells to differentiate towards the myocardial cells, and nearly 80% of myocardial cells can be obtained on the seventh day of induced differentiation. Compared with a traditional in-vitro myocardial differentiation induction scheme, the induced differentiation method provided by the invention has the advantages that the variety of related small molecules is few, the dosage is low, the induced differentiation time can be effectively shortened from 15 days to 7 days, the induced differentiation cost is saved, and the method is suitable for large-scale popularization and application. A novel way which is more economical, simpler, more convenient, more stable and more efficient is provided for the production of the stem cell-derived myocardial cells; the cardiac muscle cells obtained by induced differentiation are relatively high in purity and relatively wide in application prospect, and are of great significance to research on drug screening, myocardial regenerative medicine and cardiac developmental biology.

The invention relates to a culture solution for recovering the quality of porcine oocytes by adding uridine. The culture solution comprises a basic culture solution and uridine, the basic culture solution contains 10% of pig follicular fluid, 0.57 mM of cysteine, 0.5 IU / mL of LH, 0.5 IU / mL of FSH, 10 ng / mL of epidermal growth factors, 1% of a double-antibody TCM 199 culture medium and the balance of water; the addition amount of uridine is 100 [mu] M of the basic culture solution. According to the method disclosed by the invention, metabolite uridine (Urd) required by a human body is used for respectively rescuing in-vitro cultured H2O2 induced oxidative stress porcine oocytes and delayed cultured aged porcine oocytes, so that the quality of the oocytes is improved.

The invention relates to application of pirfenidone in preparation of testicular organs, and the pirfenidone can improve one or more properties and / or functions of the testicular organs, especially improve blood testis barriers and improve spermatogenesis efficiency. The invention also provides a method, a culture medium and a kit for preparing the testicular organoid.

The invention belongs to the technical field of biological medicines, and particularly relates to a sperm injury repair model and an establishment method thereof. Comprising the following steps: step 1, establishing a sperm lead poisoning model; and 2, establishing a lead poisoning sperm repair model. The method for establishing the lead poisoning sperm repair model in the step 2 comprises the following steps: (1) preparing a preparation for optimizing sperm functions in an extracellular vesicle way; (2) repairing the damaged sperms by using the preparation for optimizing the sperm function through the extracellular vesicles; and (3) detecting the activity of the repaired sperms. In the invention, by establishing a sperm injury repair model and adopting an extracellular vesicle modification technology, the key functional protein is introduced into the extracellular vesicle, and the method has important significance on auxiliary researches such as development and research of related drugs for improving sperm injury and improving sperm motility of asthenospermia patients in vitro.

The present invention relates to a method for culturing different types of cell in the same culture medium, to a method of culturing cells at high densities and to cell cultures comprising different types of cell.

The invention belongs to the technical field of biomedicine, and particularly relates to a preparation method of human nourishing stem cells induced by human extended potential stem cells, which comprises the following specific steps: step 1, converting H9 human embryonic stem cells into adult extended potential stem cells on the basis of a feeder-layer-free system: (1) coating culture dishes with Matrigel, adding 1ml of Matrigel working solution into 35 dishes, and uniformly stirring; placing in an incubator at 37 DEG C overnight, and fully coating the dish bottom with the substrate for later use; according to the application, H9 human embryonic stem cells are converted into adult extended potential stem cells on the basis of a feeder-layer-free system; based on a human trophoblast stem cell culture system, feeder-layer-free human extended potential stem cells are differentiated into induced human trophoblast stem cells, and finally the induced human trophoblast stem cells are identified and induced to differentiate, so that interference of feeder-layer cells on the induced trophoblast stem cells is eliminated, and the induction efficiency of the induced trophoblast stem cells under the feeder-layer-free system is ensured.

An optical arrangement receives output light emanating from an object disposed within a fluid column that crosses an optical refraction boundary of the fluid column between the object and the optical arrangement. The optical arrangement modifies the output light such that the modified output light has an intensity that is more uniform than the output light.

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 hydrogel for 3D culture of stem cells / organoids and a preparation method of the hydrogel, and belongs to the technical field of biological materials. The hydrogel for 3D culture of stem cells / organoids is formed by a four-arm polyethylene glycol macromolecular polymer, the polyethylene glycol macromolecular polymer has a maleimide group (PEG-4MAL) at each terminal, is coupled with an adhesive peptide containing cysteine, and is obtained by cross-linking with mercapto polyethylene glycol (PEG-SH). The hydrogel is beneficial to organoids, the stem cells are high in growth rate and short in culture period, and the obtained organoids are high in maturity, uniform in size, high in bionic capacity and high in application value. Therefore, the prepared novel hydrogel material provides a more ideal microenvironment for culture of cells and organoids, and promotes application and development of regenerative medicine and tissue engineering.

The invention relates to the field of cell biology, and particularly discloses a method for preparing mesoderm progenitor cells from human pluripotent stem cells and an induced differentiation culture medium of the mesoderm progenitor cells. The method comprises the following steps: inoculating human pluripotent stem cells in a single cell state into a culture dish covered with matrigel; and inducing the human pluripotent stem cells to be differentiated into the mesoderm progenitor cells within 1-3 days by using the mesoderm progenitor cell induced differentiation culture medium. Wherein the mesoderm progenitor cell induced differentiation culture medium is composed of a basic culture medium and an additive, and the additive contains a B-27 additive, non-essential amino acid, glutamine, ascorbic acid, polyvinyl alcohol, a GSK-3 inhibitor and a CDK8 inhibitor. According to the method, the human pluripotent stem cells are subjected to induced differentiation by using the mesoderm progenitor cell induced differentiation culture medium with specific composition, the method does not depend on cloning of the pluripotent stem cells, a large number of high-uniformity mesoderm progenitor cells can be obtained through induced differentiation within 1-3 days, the induction efficiency is high, and the stability is high.

The invention relates to application of an NR1D1 gene in regulation and control of granular cells, and belongs to the technical field of biology. The invention provides an application of an NR1D1 gene in regulation and control of granulosa cells, including an application of a sequence shown in siRNA of the NR1D1 gene in preparation of a product for regulation and control of sheep granulosa cells, and the sequence is shown in SEQ ID NO: 1 and SEQ ID NO: 2. According to the regulation method and product of the granulosa cells, the proliferation apoptosis and activity of the granulosa cells can be effectively regulated, and the expression of follicular development related genes can be regulated. The method has the beneficial effects that in-vivo or in-vitro regulation and control effects on follicles and / or ovaries are realized, and a method and a product guarantee are provided for research on development and physiological and biochemical functions of granular cells, follicles and ovaries.

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 discloses a pig sperm specific antibody. The coding sequence of the antibody is shown as any one of SEQ ID No: 1-3. According to the method, the nano antibody library is prepared by immunizing alpaca with whole pig sperms, and then the antibody capable of being specifically combined with the X or Y sperms can be screened out through the library, so that sorting of the X and Y sperms is realized. The screened antibody is a nano antibody, the size of the nano antibody is only 10% of that of a traditional antibody, the structure is simpler, the advantages of high stability, strong specificity and low immunogenicity are achieved, large-scale expression of the antibody can be achieved, the production cost of the antibody is greatly reduced, and industrial application is facilitated.

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 present teachings provide for a method of breeding livestock in vitro. Provided are steps to create embryonic stem cells from a plurality of blastocysts, genotype the embryonic stem cells to select the best embryos for mating to create offspring, and induce the embryonic stem cells into primordial germ cell-like cells (PGCLCs). Male PGCLCs are further induced into spermatogonial stem cell-like cells, and then spermatid-like cells. Female PGCLCs are induced into oocytes, which are then matured. The resulting gametes can then be mixed with each other or with opposite sex gametes from animals with desirable genetics to create the next generation of embryos, which can then be run through the process again. This method of in vitro breeding can be used to increase the speed of genetic progress in livestock.

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.

The invention belongs to the field of stem cell biology, and relates to pedigree specific differentiation of human pluripotent or embryonic stem cells, in particular to a mesenchymal stem cell induced differentiation culture medium and application thereof. The mesenchymal stem cell induced differentiation culture medium is used for directly inducing and differentiating human induced pluripotent stem cells / or embryonic stem cells into mesenchymal stem cells, the culture medium comprises a mesenchymal stem cell basic culture medium and an additive, and the additive comprises the following components: 2-10 [mu] M of a TGF-beta inhibitor, 2-10 [mu] M of a GSK3 beta inhibitor and 5-10 [mu] M of a Rock inhibitor. By adopting the induced differentiation culture medium provided by the invention, human induced pluripotent stem cells / or embryonic stem cells can be directly induced and differentiated into mesenchymal stem cells, and the obtained mesenchymal stem cells have the advantages of infinite sources, stable quality, high homogeneity and the like; and the human induced pluripotent stem cells / or embryonic stem cells can be directly and directionally differentiated into the mesenchymal stem cells in a relatively short time, so that the culture time is shortened, and the production cost is reduced.