139 results about "Blastocytes" patented technology

The invention discloses an image detection and evaluation method for an unfrozen form of a fertilized egg embryo. The method comprises the following steps: acquiring a time sequence image sequence after the fertilized egg embryo is unfrozen; performing image standardization processing on the time sequence image sequence to generate a standardized embryo image set; extracting multi-dimensional embryo quality parameters from the standardized embryo image set; wherein the multi-dimensional embryo quality parameters comprise a morphological static parameter set, a dynamic change parameter set and a texture feature parameter set; generating a blastocyst formation rate based on the multi-dimensional embryo quality parameters; outputting a third-level embryo quality evaluation result according to a comparison relationship between the blastocyst formation rate and a preset threshold interval; wherein the three-stage embryo quality evaluation result comprises an excellent embryo grade, a qualified embryo grade and an abnormal embryo grade. The method has the following advantages and effects: full-automatic and multi-dimensional quantitative evaluation of the thawing process of the unfrozen embryos is realized, so that the scientificity of embryo screening and the success rate of clinical pregnancy are improved.

The invention belongs to the field of antibody development, and discloses a screening method and application of a human-mouse protein high homologous target antibody based on a fully humanized antibody mouse. Aiming at the problem of weak antibody response caused by immune tolerance of human-mouse high homologous targets (protein homology is greater than or equal to 95%), the following scheme is provided: in embryonic stem cells (ES cells) of HUGO-Mabfully humanized antibody transgenic mice, a mouse target gene (such as ACVR2A) is knocked out through a Turbo Knockout technology, and homozygous knockout ES clones are screened; carrying out microinjection on the clones to the whitened B6 mouse blastocyst, and transplanting a pregnant mouse to obtain a Founder mouse; the Founder mouse is subjected to target antigen immunization for more than or equal to 4 times (the Freund's complete adjuvant is used for the first time), and the titer of the serum antibody is detected. According to the invention, 100% homozygous knockout chimeric efficiency is realized in the Founder stage, the mouse construction period is shortened from traditional 8-10 months to 3-4 months, and the diversity and affinity of the antibody are significantly improved (titer reaches 1: 729,000). The obtained antibody can be used for preparing medicines for treating tumors or autoimmune diseases.

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

The invention relates to a method for inducing mouse trophoblast stem cells in vitro. According to the method, mouse embryonic stem cells are used as starting cells, Sorcs3 gene knockout mouse embryonic stem cells are obtained through a gene editing technology, and in-vitro induced trophoblast stem cells (iTSCs) are efficiently and rapidly obtained by means of a flow cytometry sorting technology and in combination with specific antibody sorting. According to the invention, the Sorcs3 gene is knocked out from the embryonic stem cell, so that the capability of the embryonic stem cell to differentiate to the trophoblast lineage can be obviously enhanced; an important research platform is provided for deeply researching a lineage intertransformation mechanism of cell masses (ICM) and trophoblast (TE) in the blastocysts, and the method has important significance for researching lineage limitation of early embryonic development of mammals and the like.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a glucose slow-release structure used in an embryo in-vitro culture solution, a preparation process and a use method. The glucose slow-release structure comprises at least one inner layer containing glucose and a biodegradable material outer layer wrapping the inner layer, and the biodegradable material outer layer comprises a matrix with a plurality of micropores and at least one sealing material layer used for sealing pores of the micropores. According to the invention, gradient release from low sugar in the cleavage stage to high sugar in the blastocyst stage is realized, midway liquid change or artificial sugar supplement is not needed, the operation is obviously simplified, and pollution and mechanical stress risks are reduced.

The invention discloses a bovine oocyte in-vitro maturation culture solution as well as a preparation method and application thereof, and belongs to the technical field of animal reproduction and embryo engineering. The preparation method of the bovine oocyte in-vitro maturation culture solution comprises the step of adding mitoquinine mesylate into a conventional IVM culture solution, wherein the adding concentration of the mitoquinine mesylate is preferably 50 nmol / L. The bovine oocyte in-vitro maturation culture solution provided by the invention can effectively reduce the oxidative stress reaction of the oocytes in the in-vitro culture process, enhance the mitochondrial function and remarkably improve the bovine oocyte in-vitro maturation quality and the embryonic development potential, so that the blastocyst rate is increased, and the problem that the development potential of the existing oocyte IVM culture solution is insufficient is solved.

The present disclosure relates to methods, products, and compositions useful for deriving bovine induced blastoids. Methods are described that allow for juvenile stem cell aggregation and induced blastocyst formation. The invention further provides an aggregation culture medium and an induced blastocyst-like culture medium which are suitable for promoting aggregation of the young stem cells and preparing the induced blastocyst-like. The bovine induced blastoids can be used in selection and breeding procedures, including the replication of pre-implant embryos with desired characteristics and in vitro breeding including a cross-generation breeding cycle from parent germ cells and / or embryos to progeny embryos.

Described are small molecule cocktails that enable de novo derivation and long-term culture of bovine extraembryonic endoderm cells (bXENs). Methods of using the small molecule cocktails to generate bXENs are also described. Methods of using the bXENs to form blastocyst models or to maintain the stemness of bovine ESCs and prevent them from differentiation are also described.

The invention relates to the technical field of livestock and poultry breeding, in particular to a goat hybridization breeding method capable of achieving complete-cycle nutrition enhancement. The core of the method is that systematic nutrition intervention runs through the whole breeding process. Before hybridization, performing differential nutrition pretreatment on the parents for 60 days: feeding the female parents with a breeding nutrition enhancer added with components such as D-chiro-inositol and N-acetylcysteine, and supplementing sperm motility enhancer containing acetylated L-carnitine and ergothioneine to the male parents. In the gamete treatment stage, the oocytes are subjected to in-vitro maturation culture in a special culture solution containing components such as D-chiro-inositol and a growth differentiation factor 9; after the sperms are subjected to gradient centrifugal screening, completing in-vitro fertilization in an optimized fertilization culture solution; after the embryos develop into blastocysts in the sequential culture system, carrying out laparoscopic transplantation, and carrying out staged precise nutrition culture on the pregnant ewes. According to the method, through multi-level nutrition enhancement, the reproductive capacity of the goats and the growth performance of offspring are effectively improved.

The invention discloses a method for improving developmental potential of cloned embryos through high expression of Smarca5 and application of the method. According to the method, after the SCNT embryo is reconstructed, the proportion of 4 cell periods, the blastocyst formation rate and the proportion of inner cell clusters are remarkably increased by microinjection of Smarca5mRNA (500ng / mL). Immunofluorescence and qPCR results show that the expression of EGA related genes such as Polr2a and Dppa2 is up-regulated through high expression of the Smarca5, the synthesis level of new RNA is enhanced, and the abundance of H3K9me3 is remarkably reduced by adjusting the balance of Kdm4b / Suv39h2. According to the method, the embryo epigenetic reprogramming level is improved, the SCNT development quality is remarkably improved, and the method can be widely applied to the fields of animal cloning and embryo engineering.

It is revealed that an organ such as pancreas can be regenerated by utilizing a fact that the deficiency of an organ is complemented by injecting an induced pluripotent stem cell (iPS cell) into a developed blastocyst in a blastocyst complementation method. Thus, the present invention has solved the above-described object. This provides a method for producing a target organ, using an iPS cell, in a living body of a non-human mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a different individual mammal that is an individual different from the non-human mammal.

The present invention relates to a medium composition for in vitro fertilization and / or in vitro culture comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, and a method using same. The compound or composition suppresses reactive oxygen species and lipid peroxidation in ovarian granulosa cells to preventing cytotoxicity and mitochondrial dysfunction caused by apoptosis and ferroptosis, leading to an improvement in the quality of aged oocytes, and to increase the developmental rate and blastocyst formation rate of pre-implantation embryos, thereby improving the efficiency of in vitro fertilization. In addition, treatment with the medium composition during the vitrification process of in vitro–fertilized embryos improves re-expansion and survival rates, thereby reducing structural and metabolic damage. Accordingly, the present invention can be advantageously applied to an assisted reproductive technology for treating infertility and subfertility, and in vitro fertilization and / or in vitro culture for improving the propagation efficiency of livestock.

The invention provides an oocyte in-vitro maturation culture solution additive and application thereof. The additive is tert-butylhydroquinone. By adding 100 [mu] g / mL of TBHQ into a basic culture solution, the first polar body discharge rate of oocytes is remarkably increased to 77.29%, the ROS level is reduced, and the blastocyst rate of in-vitro fertilized embryos is increased. The technology is suitable for breed conservation of livestock genetic resources such as sheep and can replace live livestock transportation to reduce the cost.

The invention belongs to the technical field of bioengineering, and particularly relates to application of RPS26 recombinant protein in promoting early embryonic development of animals. Experiments show that after the RPS26 recombinant protein is added in the early-stage in-vitro embryo culture process of the lamb goats, the blastocyst rate of the lamb goats can be increased to 40.0%; meanwhile, the cell proportion of cell mass in the blastocyst can be remarkably increased, the cell proportion of trophoderm is reduced, and the total number of blastocyst cells is not influenced, so that the blastocyst development rate can be remarkably increased and the blastocyst quality can be improved by utilizing the RPS26 recombinant protein.

The embodiment of the invention discloses an in-vitro fertilization embryo culture blastocyst formation rate prediction method and system. The method comprises the following steps: acquiring parameter information of an in-vitro fertilization embryo to be evaluated; the parameter information serves as input and is input into a pre-trained prediction model, a formation rate prediction result of the to-be-evaluated in-vitro fertilized embryo is obtained, and the pre-trained prediction model is obtained through training based on a LightGBM model; and if the formation rate prediction result of the to-be-evaluated in-vitro fertilized embryo is greater than a preset threshold value, using the to-be-evaluated in-vitro fertilized embryo for subsequent transplantation.

The present invention belongs to the field of molecular biology and genetics, and in particular relates to a method for editing a bovine gene based on Pro-iCHI. The present invention provides a method for editing a bovine gene based on Pro-iCHI. Protamine is transiently expressed in gene-edited b-haSCs, which are then injected to mature oocytes to obtain reconstructed embryos. Protamine can eliminate abnormal DNA methylation resulting from oocyte intracytoplasmic haSCs injection and enable the nucli to compress into sperm-like structures, and the obtained bovine Pro-iCHI embryos can successfully develop into blastocysts, with a blastocyst rate comparable to that of the embryos obtained by in vitro fertilization. Moreover, in the present invention, a protamine-encoding gene is inserted into a Saccharomyces cerevisiae protein expression vector for transient expression, which ensures that abnormal DNA methylation is erased, without integration into the genome resulting in the insertion of exogenous genes.

The invention relates to the technical field of construction of conditional gene knockout models, and discloses a construction method of a mouse Oct4 conditional gene knockout model based on quadruple sgRNA collaborative targeting, which comprises the following steps: step 1, designing four sgRNAs, namely sgRNA1, sgRNA2, sgRNA3 and sgRNA4, in which the sgRNA1 and the sgRNA2 target 5'end and 3 'end non-coding regions of a second exon of an Oct4 gene, and the sgRNA3 and the sgRNA4 target structural domain conserved sequences; step 2, preparing an electrotransfection solution, wherein the electrotransfection solution is composed of Cas9mRNA, the four sgRNAs and an electrotransfection buffer solution; step 3, carrying out electroporation treatment on the fertilized eggs of the mice; step 4, culturing the fertilized eggs subjected to the electrotransformation treatment to a blastocyst stage; and 5, carrying out genotype identification on the cultured blastocyst. According to the scheme, the sgRNA combination and embryo electrotransfection system is optimized, the mouse Oct4 conditional gene knockout model is efficiently and accurately constructed, a reliable tool is provided for regulation mechanism research of Oct4 in mouse early embryo trigerm differentiation, and the medical transformation value of Oct4 is excavated.

A method of deriving and maintaining Salmonidae stem cells is provided, the method comprising deriving Salmonidae cells at the eyed or blastula stages of salmonid or salmonoid ova development, so as to produce cells with use in the food industry.

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.

This invention discloses a method and system for embryo quality detection, relating to the field of embryo detection technology, including the following steps: S1: fertilized embryos are continuously cultured in an in vitro culture system containing specific metabolic marker detection components, and continuous dynamic imaging of the embryos is performed to obtain a complete dynamic developmental image sequence; S2: the dynamic developmental image sequence is segmented, and temporal developmental features are captured, identifying key time points such as pronucleus appearance and disappearance, cleavage, and blastocyst formation, and generating a segmentation mask corresponding to each frame of the dynamic developmental image sequence; this invention achieves full-process time-difference imaging to capture the temporal features of embryonic development, without trauma or interference, avoiding subjective bias in static observation; by segmenting the dynamic developmental image sequence and capturing temporal developmental features, the segmentation accuracy is high and the anti-interference ability is strong.

This invention discloses a culture medium additive for improving embryonic developmental potential and its application, belonging to the field of in vitro embryo culture technology. This invention is the first to discover that CPI-455, a KDM5 histone demethylase inhibitor, can significantly improve the blastocyst rate, developmental speed, and embryo quality of preimplantation embryos by increasing H3K4me3 levels within a certain concentration range. Simultaneously, the addition of CPI-455 and NMN can substantially increase the rate of high-quality / high-quality blastocysts and embryo quality in in vitro embryo culture. The embryonic developmental potential additive provided by this invention is of great significance for improving in vitro embryo culture efficiency and embryo quality, accelerating the process of breeding superior embryos, and assisting in research on embryo-related mechanisms.

The present application discloses an artificial propagation-hybridization combined breeding method for meat goats, which relates to the technical field of animal breeding. The specific steps include: selecting a goat breed with good meat performance as the maternal parent and selecting a goat breed with a large body as the paternal parent; culturing the maternal oocytes in vitro to mature oocytes containing the first polar body, using a temperature gradient screening device to screen out highly motile sperm, fertilizing the mature oocytes with the highly motile sperm in vitro, culturing them to a hybrid blastocyst morphology, transplanting them into the maternal body to develop until natural delivery, and obtaining a hybrid F1 generation; backcrossing the F1 generation with the maternal parent, and backcrossing the backcross offspring with the maternal parent multiple times until the breeding goal is met, and then cross-crossing and fixing them to obtain a new breed / line with excellent traits; improving the sperm screening efficiency by utilizing the screening device, ensuring the quality of the screened sperm, and thus significantly improving the fertilization efficiency and embryo survival rate.

This invention relates to the field of bioengineering technology, and in particular to a method for simultaneously collecting the inner cell mass and trophectoderm of mammalian blastocysts. The method includes the following steps: (1) after removing the zona pellucida of the blastocyst, the blastocyst is cut into an inner cell mass and a trophectoderm portion, and the cut trophectoderm portion is transferred to a culture medium for collection; (2) the inner cell mass portion obtained in step (1) is sequentially placed in an antibody treatment solution and a complement treatment solution for incubation; (3) the inner cell mass portion treated in step (2) is pipetted to obtain a purified inner cell mass. This method achieves simultaneous and efficient separation of two key cell lines from the blastocyst, fundamentally changing the traditional method's "resource waste" model of only obtaining the inner cell mass while discarding the trophectoderm, and providing more comprehensive experimental materials for embryonic development, placental biology, and stem cell research.

The present application relates to the field of animal embryo engineering technology, and provides a method for improving the development ability of bovine IVP embryos. In the production of bovine in vitro embryos, by adding PAPPA, ergothioneine and selenocysteine to the bovine oocyte in vitro maturation culture solution, early embryo culture solution, late embryo culture solution, freezing solution and thawing solution, the in vitro maturation performance of oocytes and the development performance and vitality of the subsequent obtained embryos can be effectively improved. Studies have shown that after adding the above additives, the maturation rate of oocytes, the cleavage rate, the blastocyst rate of embryos, and the freezing survival rate, the hatching rate and the ATP content of embryos after freezing preservation are significantly improved, which has important application value.

The application relates to a mouse model construction method for psoriasis research based on SBSN 600 lysine dihydroxyisobutyryl modification, which comprises the following steps: first step, constructing a plasmid comprising a DNA fragment of a SBSN gene sequence, wherein 600 sites of the SBSN sequence are mutated from lysine (K) to threonine (T); second step, culturing mouse embryonic stem cells; third step, electrically transforming the embryonic stem cells; fourth step, drug screening; fifth step, culturing the embryonic stem cells screened in the fourth step; sixth step, injecting the embryonic stem cells cultured in the fourth step into mouse blastocysts, then culturing the mouse blastocysts, and transplanting the cultured blastocysts into the uterus of a pseudopregnant female mouse, wherein the mouse delivered by the female mouse is a chimera mouse; and seventh step, breeding the chimera mouse and identifying the mouse genotype through PCR.

The invention relates to the technical field of artificial intelligence in assisted reproduction, in particular to an embryo optimization model construction method, device and equipment and a storage medium. Comprising the following steps: acquiring video data of embryonic development from a plurality of fertilized eggs to a blastocyst stage based on a delayed photography embryo culture technology; constructing time sequence image data according to the video data; the time sequence image data comprises multiple frames of continuous images; performing feature extraction on each frame of image of the time sequence image to obtain a high-dimensional feature corresponding to each frame of image; determining a comparison difference feature with the corresponding reference sample feature, performing fusion, and constructing a fusion feature corresponding to each frame of image; constructing a feature sequence of the target embryo development according to the fusion features; and acquiring a trained embryo quality grading model according to a time dependency relationship of the feature sequence for capturing embryo development. The model provided by the invention can optimize the utilization rate of embryos, reduce unnecessary repeated ovulation promoting cycles, and promote the development of reproductive medicine towards the precision direction.

The application provides a blastocyst three-dimensional fitting reconstruction method and device, electronic equipment and storage medium. The method comprises the following steps: acquiring a plurality of focal plane images of a blastocyst; identifying a trophoblast cell mask and an inner cell mass mask in the blastocyst based on the plurality of focal plane images; constructing a trophoblast cell grid model and an inner cell mass grid model according to the trophoblast cell mask and the inner cell mass mask; constructing a depth map of the blastocyst according to the plurality of focal plane images; performing image fusion on the plurality of focal plane images to obtain a texture image; performing plane mapping according to the texture image and the depth map to generate an internal three-dimensional point cloud model of the blastocyst; constructing a three-dimensional geometric model of the blastocyst according to the trophoblast cell grid model, the inner cell mass grid model and the internal three-dimensional point cloud model; and mapping the texture image to the surface of the three-dimensional geometric model to generate a blastocyst three-dimensional model. The reconstruction based on the method does not affect the development of the embryo, and is low in cost and safe and reliable.

This invention relates to the fields of biomedical engineering and embryo quality assessment, and particularly to a biomarker for assessing the quality of IVM-derived embryos and its application. The biomarker includes time point parameters t2 and t9 of the IVM-derived embryos, and time interval parameters S2, t9–t7, t9–t8, or tSB–t9. A method for assessing the quality of IVM-derived embryos based on the aforementioned biomarker has been established. This method combines non-invasive temporal imaging technology with morphodynamic multi-parameter analysis, providing a reliable tool for assessing the blastocyst formation potential and chromosomal euploidy of IVM-derived embryos in clinical practice, thus potentially expanding the application of IVM technology in assisted reproductive technology.

This invention provides a method for the cryopreservation and reconstitution of porcine embryos. During the in vitro maturation of porcine oocytes, porcine IVF embryo culture, and embryo freezing and thawing processes, the addition of TSA, 5-Aza-dC, Etomoxir, GSK-J4, and NR, either alone or in combination, improves the development and freezing efficiency of porcine IVF embryos. Experiments show that the oocyte maturation rate, embryo cleavage rate, and blastocyst rate of the groups adding TSA, 5-Aza-dC, and Etomoxir to the porcine oocyte in vitro maturation culture medium and porcine IVF embryo culture medium are significantly higher than those of the control group. Furthermore, the hatching rate of porcine IVF embryos after freezing and thawing is significantly higher in the groups adding GSK-J4 and NR to the freezing and thawing solutions than in the control group. This invention lays the foundation for the production of porcine in vitro fertilized embryos and related scientific research, providing an effective, safe, and feasible strategy for improving the production efficiency and quality of porcine in vitro embryos.

The invention discloses a culture solution for improving the development rate of in-vitro cultured pig blastocysts as well as application and a use method of the culture solution, and belongs to the technical field of livestock embryo engineering. The invention discloses a culture solution for improving the development rate of in-vitro cultured pig blastocysts, which is characterized in that a PZM3 culture solution is used as a basic culture solution, and 5-25ng / mL of tumor necrosis factor superfamily cell factor TNFSF12 is added. According to the present invention, the pig embryo needs to be subjected to in-vitro culture for 4 days in the TNFSF12-free basic culture solution, and then is transferred to the TNFSF12-containing basic culture solution so as to be continuously cultured for 2 days, such that the generation efficiency of the pig blastocyst can be effectively improved, and the support is provided for the application of the pig in-vitro fertilization, the somatic cell nuclear transfer, the embryo genome selection and other embryo engineering technologies in the agricultural breeding and the human medicine;