62 results about "Embryo cell" patented technology

The invention provides biopsy liquid and application thereof in pig blastocyst laser cutting, and belongs to the technical field of animal biology. The invention provides a biopsy solution and a preparation method thereof, the biopsy solution can be applied to pig blastocyst biopsy by laser cutting, the problem that at present, no adaptive biopsy solution exists for laser cutting of pig blastocyst is solved, and the invention further provides a method for laser cutting of pig blastocyst by using the biopsy solution. The problems of embryo cell injury, embryo development potential damage and the like possibly caused by laser cutting of the pig blastocyst in the prior art are solved. The biopsy liquid and the use method thereof can be efficiently and accurately applied to acquisition of pig blastocyst trophoblast cells, and are low in cost and good in repeatability.

The invention provides an improved blastocyst culture solution as well as a preparation method and application thereof, and relates to the field of embryonic cell in-vitro culture. Based on a formula of a conventional blastocyst culture solution, colony stimulating factor-1 (CSF-1) and a transforming growth factor-alpha (TGF-alpha) are added, and the colony stimulating factor-1 and the transforming growth factor-alpha are dissolved in the conventional culture solution according to the concentration of 0.02-0.06 mmol / L and the concentration of 0.01-0.05 mmol / L; the obtained improved blastocyst culture solution can improve the mulberry embryo formation rate or blastocyst formation rate in in-vitro culture, promote the development of low-quality embryos and improve the mulberry embryo quality or blastocyst quality, and can be used for preparing medicines for promoting embryo implantation or subsequent growth and development.

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 precise clamping type multi-station embryo culture dish operation table and a use method thereof, and relates to the technical field of embryos. A guide groove is formed in the inner side of a guide frame, a plurality of alignment flat grooves are formed in the top side face of the guide frame, and a plurality of spiral fasteners aligned with the alignment flat grooves are installed on the side face of the guide frame in a screwed mode. The clamping tooth piece comprises a toothed plate, two vertical plates fixed to the upper side of the side end of the toothed plate and a transverse rod fixed between the upper side ends of the two vertical plates, and a sliding piece installed at the guide groove in a sliding mode is further fixed to the side end of the toothed plate. The embryo culture dish comprises a lower ring body and an upper ring body, and a ring gap is formed between the periphery of the lower ring body and the upper part of the upper ring body. The gear ring piece comprises a ring plate which is positioned and sleeved at the position of the ring position notch; and an outer ring tooth opening matched with the side tooth opening is formed in the outer ring side of the ring plate. The culture dish is effectively prevented from being mistakenly touched and shaken, the problem that normal development of cells is affected by a high-intensity magnetic field and the like is also avoided, and a stable external environment is provided for embryonic cell culture.

This invention relates to the field of embryo resuscitation technology and discloses an auxiliary warming device for frozen embryo resuscitation, including a collection plate. A fixing ring is fixedly connected to the side wall of the collection plate, and a forward threaded groove is formed on the side wall of the fixing ring. The cover plate drives the separator ring to rotate clockwise through the contact rod and the fixing plate. Since the reverse threaded groove rotates in the opposite direction to the forward threaded groove, the rotating ring will move upward at this time. The upward rotating ring will drive the separator ring to move upward synchronously, so that the through hole gap coincides with the flow hole. At this time, the new solution on the outer wall of the separator ring flows downward through the flow hole and the through hole gap. Finally, the new solution mixes with the solution inside the collection plate. At this time, the pressure of the dripping will be absorbed by the top of the fixing ring and will not directly contact the solution inside the collection plate. When the operator twists the cover plate to close it, the flow hole and the through hole gap will coincide, and the new solution will mix with the solution inside the collection plate, effectively reducing the damage to the outer wall of the embryo cell membrane when injecting the new solution.

The utility model relates to an embryonic cell culture kit based on gelatin, which is applied to the technical field of reagent packaging boxes, and realizes that when a reagent in a kit body is used, after a bottle cap is opened, a handle is rotated to be in a vertical state, and then the handle is held by a hand to drive a turning cover to tear a rubber plug, so that the reagent can be taken out from a bottle opening, and the operation is convenient. After the reagent is taken out, the rubber plug can be pressed back to the original position to make the rubber plug abut against the clamping box on the inner wall of the bottle opening, so that the sealing performance is ensured, then the handle is rotated to be in the horizontal state with the turning cover, and at the moment, the reagent box body can be refrigerated and stored after the bottle cap is in threaded connection with the bottle opening; after the turning cover and the rubber plug are turned over and opened, the extending opening is clamped at the top end of a bottle opening, the rubber plug is pressed, then a reagent can be easily poured out through the leaking opening, the dripping leakage phenomenon is avoided, and the effects that the unsealed reagent is conveniently used, the sealing performance is improved, and storage is convenient are effectively achieved.

The invention provides genetically modified non-human animals that express chimeric human / non-human MHC I polypeptide and / or human or humanized β2 microglobulin polypeptide, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified animals and methods of making the same. Methods of using the genetically modified animals to study various aspects of human immune system are provided.

The present application relates to a primer combination for amplifying whole genome DNA of sheep embryo cells and application thereof, and the primer combination is composed of 20 or 30 primers. The present application designs random primers based on sheep reference genome sequence, optimizes the reaction system based on MDA whole genome amplification technology, so that the whole genome DNA amplification product with high coverage and low mismatch rate can be generated by using 10 sheep embryo cells or as low as pg level genomic DNA, which can be directly adapted to downstream applications such as second-generation sequencing and targeted capture sequencing, and the detection cost is reduced.

The invention relates to the technical field of embryonic development evaluation, and discloses an embryonic chromatin openness and gene expression evaluation method, which comprises the following steps: extracting an embryonic cell sample with a survival rate of more than or equal to 95% through a micromanipulation technology; acquiring chromatin open signal data by adopting an ATAC-seq technology; carrying out genome positioning by utilizing a BWA comparison tool, and constructing a three-dimensional contact matrix through a matrix decomposition algorithm in combination with Hi-C data; determining the gene expression quantity by adopting fluorescent quantitative PCR (Polymerase Chain Reaction); and constructing an XGBoost integrated learning model to fuse chromatin characteristics and gene expression data. By optimizing micromanipulation parameters and an ATAC-seq experimental process, the cell survival rate is increased to 95.2 + / -1.3%, the ATAC-seq library complexity reaches 85.3%, and Hi-C data standardization processing is combined, so that the technical problem of data deviation is solved, and the embryo sample processing efficiency and data reliability are remarkably improved.

The invention provides a chromatin conformation capturing method of high-lipid-droplet-content single cells based on low-melting-point agar immobilization, and relates to the technical field of chromatin conformation capture.The chromatin conformation capturing method comprises the steps that permeabilized single cells are placed in a buffer solution, and an isometric low-melting-point agarose solution is added to immobilize the cells; the mass concentration of the low-melting-point agarose solution is 0.5% w / v. According to the method, permeabilized cells are fixed by adopting low-melting-point agarose, the problem that cell nucleuses of high-lipid-droplet-content cells are not easy to track in chromatin conformation capture of the cells is solved, the influence of lipid droplets on enzyme digestion of the cell nucleuses is reduced by adopting a 0.5% w / v low-melting-point agarose solution, and the total chromatin interaction number obtained by data is superior to that obtained by adopting a single-cell Hi-C method in the prior art; compared with the bulk Hi-C data of the same type of embryonic cells, the same variation trend and interaction distribution can be displayed. The technical problem that in the prior art, a common single cell Hi-C technology is not suitable for high-fat-content single cells of pig embryos is solved.

The invention relates to the technical field of agricultural biology, in particular to a method for increasing the germination rate of hemerocallis citrina L. seeds, the method for remarkably increasing the germination rate of the hemerocallis citrina L. seeds through combined treatment of gibberellin GA3 and hydrogen peroxide H2O2 is easy and convenient to operate and low in cost, the physiological dormancy of the seeds is broken through gibberellin, embryo cell division is promoted, and the germination rate of the seeds is increased. Meanwhile, hydrogen peroxide is used for moderately oxidizing seed coats, permeability is improved, active oxygen signal molecules are provided for activating germination related enzyme systems, the hydrogen peroxide and the active oxygen signal molecules generate a synergistic interaction effect through a dormancy breaking-permeation promoting-activation triple mechanism, the germination rate is increased in a breakthrough mode, and the germination period is remarkably shortened.

The application discloses a single-cell transcriptome data pseudo-time trajectory analysis method and a computer system, which comprises the following steps: 1) calculating a gene explicit comparison advantage matrix; 2) obtaining a gene similarity matrix by similarity and constructing a gene network; 3) taking an initial node in the gene network, starting random walking from the initial node, recording each gene walked through to form a gene text composed of gene sequences; 4) converting the gene text into a gene word vector; 5) adding all single-cell expressed gene vectors with expression as weight to form a sum vector as a word vector representation of the single cell in the gene space; and 6) visualizing all cell vector representations to obtain an embryo cell development pseudo-time trajectory result. The application provides an analysis basis for identifying different rare cell subtypes in tissues and variant genes of different cell subtypes and has a wide and important application prospect in the fields of tumors, developmental biology and life science.

The present invention relates to an exogenous gene delivery system based on carbon dot nanoparticles. The preparation method of the carbon dot nanoparticles of the present invention is to electrolyze a graphite rod with a voltage of 15V - 20V to obtain an electrolyte solution, filter and dialyze the electrolyte solution to obtain a carbon dot solution; add an activator to the carbon dot solution for the first ice-water bath reaction, then add polyethyleneimine and carry out the second ice-water bath reaction, and obtain carbon dot nanoparticles after dialysis. Mixing the carbon dot nanoparticles of the present invention with a vector carrying an exogenous gene can prepare the exogenous gene delivery system of the present invention, and transgenic plants can be achieved by coating or soaking; moreover, the carbon dot nanoparticles of the present invention are tubular, so the exogenous gene delivery system can enter the seed embryo cells and carry the exogenous gene when the plant grows, and transgenic plants can be achieved from the root source.

The application discloses an electroporation transfection protection fluid suitable for abalone embryo cell gene editing and application thereof, and belongs to the technical field of gene editing. The electroporation transfection protection fluid comprises a basic fluid and a cell protection agent; the basic fluid comprises NaCl, MgSO4, CaCl2, KCl, NaHCO3, NaBr, K2HPO4, glucose and glutathione; and the cell protection agent comprises polyethylene glycol 400. The electroporation transfection protection fluid can significantly reduce the deformity rate and mortality rate of abalone cells after electroporation technology, improve the gene transfection and gene editing efficiency, and reduce the cost of gene editing experiments, thereby laying a foundation for accelerating marine biological research and promoting environmental protection, species conservation and aquaculture.

Methods of producing synthetic embryos are provided. Accordingly, a method of producing a synthetic embryo is provided, comprising inducing expression of a factor in a subset of primitive pluripotent stem cells (PSC) that induces differentiation into trophoderm cells to obtain trophoderm originating cells; inducing expression of factors which are induced to differentiate into extraembryonic primitive endoderm cells in the second subgroup of the primitive state PSC to obtain extraembryonic primitive endoderm originating state cells; and mixing the trophoblast ectoderm originating state cells and the extraembryonic primitive endoderm originating state cells with primitive state PSC under conditions that allow the formation of aggregated cells. Articles, mixtures, and aggregates of cells and methods of using the same are also provided.

The invention provides a pelteobagrus fulvidraco embryonic cell line as well as a subculture method and application thereof, and belongs to the field of fish cell biology. The pelteobagrus fulvidraco cell line is a pelteobagrus fulvidraco embryo-derived cell line and is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C202516. The pelteobagrus fulvidraco embryonic cell line YCE is a fibrous cell, is continuously cultured for more than 100 generations, has a good growth state, can be stably passaged, can provide a large number of pelteobagrus fulvidraco embryonic cells, can be cryopreserved, and can still be stably passaged after being unfrozen. The fish embryo source cell line disclosed by the invention not only can be used for related research on an anti-infection immune mechanism of pelteobagrus fulvidraco, but also provides a new thought for an establishment method of a freshwater fish embryo cell line, and enriches basic biological data of fishes.

The invention provides genetically modified non-human animals that express chimeric human / non-human MHC I and MHC II polypeptides and / or human or humanized β2 microglobulin polypeptide, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified animals and methods of making the same. Methods of using the genetically modified animals to study various aspects of human immune system are provided.

This invention discloses a kit for amplifying the whole genome of bovine embryonic cells and its applications. The invention provides a primer set for amplifying the whole genome of bovine embryonic cells, consisting of 20 primers, each 6 nt in length. This invention can utilize 4-8 bovine embryonic cells or genomic DNA down to the pg level to amplify and generate whole-genome DNA amplification products with high coverage and low mismatch rate. These products can be directly adapted to downstream applications such as next-generation sequencing and targeted capture sequencing, while reducing detection costs. Furthermore, the three-step operation process of this invention (cell lysis-lysis termination-isothermal amplification) can complete efficient amplification within 3.5 hours.

The utility model relates to the technical field of medical culture equipment, in particular to a positive pressure swing culture system, which comprises a closed box body, a mounting plate is fixedly mounted at one end of the bottom of the box body, a plurality of culture bottles communicated with the mounting plate are fixed in the mounting plate, a gas input end is arranged on the box body, and a gas outlet end is arranged on the gas input end. The other side of the gas input end is hermetically connected with a gas washing bottle arranged in the box body through a gas pipe, and the gas outlet end of the gas washing bottle is communicated with the mounting disc through a gas pipe. By controlling and adjusting the pressure and temperature of the culture environment and driving the culture bottle to move eccentrically, the natural growth environment of the embryonic cells in a living body is simulated, so that the growth and development of the embryonic cells are promoted, and the culture efficiency of the embryonic cells is improved.

The utility model relates to the field of assisted reproductive technology equipment discloses a kind of bovine in vitro embryo culture environment simulation device, including device body assembly, the device body assembly includes simulation device body, the outer surface of the simulation device body is rotatably connected with turnover observation window, the bottom of the simulation device body is fixedly connected with buffer rod by bottom mounting seat.The utility model is through the series connection effect of first spring damper, buffer spring and second spring damper, device forms gradient shock-absorbing network in three-dimensional space, horizontal direction impact is preferentially absorbed by first spring damper and converted into heat energy dissipation, vertical load is guided to spiral spring by buffer rod and carried out flexible support, extend the action cycle to reduce peak acceleration, moving block linkage mechanism cooperates second damper to realize the vector decomposition of oblique disturbance, finally reduce the vibration amplitude at culture dish, provide nearly stationary growth interface for embryo cell division.

The invention belongs to the technical field of immunoassay, and particularly relates to application of a Tmx2 gene in assisted reproduction screening. The invention verifies that after the Tmx2 gene is knocked down in the embryo of a mouse, the proliferation of embryonic cells is inhibited, and the apoptosis of the embryonic cells is induced. The Tmx2 gene provided by the invention can be used as a marker for evaluating the development condition of an embryo, meanwhile, normal expression of the Tmx2 gene in the embryo can be ensured, the development rate of the embryo in assisted reproduction can be improved, meanwhile, a medicine for treating preimplantation embryo development retardation and recovering Tmx2 gene activation is prepared, and a new clinical scheme is provided for improving the development rate of an embryo in vitro in assisted reproduction.

The invention provides a method for improving production and freezing efficiency of in-vitro embryos of cattle and sheep, which comprises the following steps of: adding fuchsin, linoleic acid and Mg < 2 + > into an IVC culture medium, and detecting the developmental capacity, freezing survival rate, lipid content, embryonic cell apoptosis level, ROS level, cytoskeleton and the like of the embryos; the invention is helpful for establishing an effective method to reduce the lipid content of the cattle and sheep embryos, thereby improving the developmental ability of the vitrification frozen cattle and sheep embryos.

The invention discloses an application of erlotinib in preparation of an integrated drug for promoting retinal pigment epithelial cell transplantation. It is found that human embryo RPE cells (human fRPE, hfRPE) transplanted to the subretinal space of a mouse specifically express EGFR, and after erlotinib intragastric administration (40mg / kg) treatment is performed on the mouse, it is observed that under the condition that an immunosuppressive agent is not used, xenogeneic donor cells can be better expanded in the subretinal space of the host and integrated into the RPE cells of the host, tight connection is formed, and therefore the immunosuppressive effect is achieved. Meanwhile, the RPE65 functional protein is stably expressed. Under the treatment of the erlotinib with the concentration, the integrity of tissue structures (retinal pigment epithelial cells, photoreceptor extracellular nodes and the like) of a subretinal space is not influenced.

The utility model relates to a sheep embryo freezing device, which comprises a box body, a bracket, a box cover and freezing tubes, the bracket is arranged in the box body, sunken grooves are uniformly distributed on the bracket, and the freezing tubes are arranged in the grooves; a box cover is arranged at the top of the box body; a cavity between the bracket and the box body is filled with a freezing medium; and the bracket is also provided with a liquid nitrogen supplementing device and a temperature detection probe. On the basis of a medium cryopreservation box, a device for detecting the temperature of the cryopreservation box and a corresponding device for supplementing liquid nitrogen to a medium to accelerate cooling of the medium are additionally arranged, and by monitoring the temperature of the cryopreservation box, the medium in the cryopreservation box is accelerated to cool in a short time in a liquid nitrogen supplementing mode when the temperature is reduced to a set low temperature; therefore, the retention time of the ice crystals in an ice crystal generating zone is reduced, and the generation of the ice crystals is reduced, namely the mechanical damage of the ice crystals to embryonic cells is reduced.

The present invention discloses a method for identifying the breeding value of dairy cow embryos. The method comprises performing genomic selection on in vitro dairy cow embryo cells to be tested, obtaining the genomic breeding value of the dairy cow embryo to be tested, and identifying the breeding value of the dairy cow embryo based on the genomic breeding value. The present invention develops a new technology for rapid detection and evaluation of genetic performance of dairy cow breeding embryos before implantation, combines embryo micromanipulation sampling technology and genomic selection technology, and performs early and accurate genomic detection and genetic evaluation of embryos. The present invention applies whole genome selection technology to early detection of dairy cow embryos for the first time, transforming the original detection after calf birth to predicting the traits of reserve cows before embryo implantation, screening high-quality embryos, and rapidly expanding high-yielding dairy cows through embryo transplantation technology, constructing a high-yielding dairy cow breeding core group and screening excellent seed cows, thereby improving the directional and qualitative seed production capabilities of dairy cows, cultivating excellent breeding cows, improving selection accuracy, shortening the breeding cycle, accelerating genetic progress, and reducing breeding costs.

The invention discloses a human protogut-like in-vitro model based on embryonic cells, a construction method and application, and belongs to the technical field of protogut-like model culture. In order to solve the problem that the formation process of human original stripes cannot be simulated in vitro, a completely defined co-culture system is developed, embryonic stem cells and various extraembryonic cell types are cultured together, the regulation and control processes of in-vivo amniotic ectoderm, trophoblast and extraembryonic mesoderm are simulated, and the formation process of the human original stripes can be simulated in vitro. The invention also discloses the effect of different extraembryonic cells, which are not known before, in adjusting embryonic cells. Besides, by utilizing the advantages of the micro-engineering technology, the interaction between different extraembryonic cells and embryonic cells is reconstructed in space and molecule, and it is proved that the formation process of human original stripes can be summarized only through coordinated regulation and control of the extraembryonic cells. And a convenient in-vitro research platform is provided for analysis of human early-stage protointestine formation.

The invention provides application of nicotinamide ribose in preparation of drugs for increasing the number of primordial germ cells and prolonging the growth cycle, and relates to the technical field of cell biology. Animal experiments prove that the supplement of nicotinamide ribose in the embryonic period can significantly increase the number of PGCs and prolong the post-birth growth cycle, and the action mechanism comprises: nicotinamide ribose promotes the proliferation of embryonic cells by up-regulating the NAD + level in PGCs, and enhances the expression of TFAP2C at the same time to maintain the undifferentiated state of cells and delay the meiosis initiation of female embryo PGCs and the differentiation initiation of male embryo PGCs; in post-birth effect evaluation, the number of follicles in ovaries of the female child rats in the NR group is remarkably increased, the fertility cycle is remarkably prolonged, the fertility of filial generations is not affected, and nicotinamide ribose is prompted to effectively maintain reproductive reserve and enhance the fertility by optimizing the development time sequence of PGCs. The invention provides a solid experimental foundation for the application of nicotinamide ribose in increasing the number of primordial germ cells and prolonging the growth cycle, and has a wide application prospect.

To provide a method for producing a population of high titer recombinant adeno-associated virus (AAV) lacking prokaryotic sequences.SOLUTION: Culturing a human embryo cell strain in suspension, transfecting the human embryo cell strain with (a) nucleic acids sequences encoding helper proteins sufficient for rAAV replication, (b) nucleic acids sequences encoding AAVrep and AAVcap genes, and (c) a closed ended linear duplex rAAV vector nucleic acids comprising at least one inverted terminal repeats (ITR) sequence and a heterologous transgene operably linked to one or more regulatory elements, incubating the transfected human cell strain for about 40 to 400 hours, and optionally, B. lysing the transfected human cell line to purify the nucleic acid sequence encoding the rAAV, thereby producing the rAAV.SELECTED DRAWING: None

The invention discloses a method and a kit for whole genome sequencing and library building of embryo trace cells, and belongs to the technical field of molecular biology. The method comprises the following steps: (1) splitting 5-16 trace embryonic cells by using a cell splitting reagent, and releasing genome DNA (Deoxyribose Nucleic Acid); (2) carrying out enzyme digestion breaking on the genome DNA to obtain a DNA fragment; (3) connecting an amplification linker sequence to two ends of the DNA fragment; (4) carrying out PCR (Polymerase Chain Reaction) amplification by utilizing a fixed primer complementary with the amplification linker sequence, and constructing a sequencing library; wherein the step (1) to the step (4) are completed in the same reaction tube. Compared with a traditional single cell amplification method, the method is shorter in time consumption, simpler and more convenient to operate and lower in cost, and is helpful for quickly promoting the early screening of human individual genetic diseases and the early selection process in livestock and poultry breeding.

The invention relates to the technical field of biology, in particular to a preparation method of mesenchymal-like stromal cells derived on the basis of cat embryonic cell spheres. According to the preparation method of the mesenchymal-like stromal cells derived on the basis of the cat embryonic cell spheres, disclosed by the invention, the cell spherical aggregate is firstly cultured, and then adherent culture and induction are performed, so that the prepared cat mesenchymal-like stromal cells are higher in quality. In addition, by optimizing the formula of the induction culture solution, the concentration of the bFGF is improved, and the cell activity and the cell quality are effectively improved.