13 results about "Zona pellucida" patented technology

An automated method for assessing the ZP binding capacity of sperm from morphological characteristics of sperm in assisted reproduction using deep learning is disclosed. The invention also provides a method for predicting fertilization success based on ZP binding capacity of sperm in clinical assisted reproduction.

The invention discloses a dairy cow early embryo sex identification method, and belongs to the technical field of dairy cow breeding and breeding. The identification method comprises the following steps: (1) culturing a dairy cow embryo to an early blastocyst stage, starting to perform single-embryo single-droplet culture, and after culturing until a blastocyst cavity is formed, punching a blastocyst zona pellucida to promote the blastocyst to shrink and release blastocyst cavity liquid; (2) collecting blastocyst cavity liquid, and extracting embryo free DNA in the blastocyst cavity liquid; (3) by taking the free DNA as a template, designing a primer to amplify a specific repetitive sequence ChrY fragment of the bull Y chromosome; if the amplification product exists, the embryo is identified as a bull, and if the amplification product does not exist, the embryo is identified as a cow. The method can ensure that the embryo free DNA in the obtained blastocyst cavity fluid is only derived from the embryo, and can effectively avoid embryo injury. According to the method disclosed by the invention, the copy number of the amplified target area in the bull Y chromosome is more than 2000 times, a detection signal can be amplified, and the detection accuracy is improved.

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.

This invention relates to the field of reproductive engineering technology, specifically to the application of rutabagastone in improving the cryopreservation and in vitro maturation of oocytes. This invention is the first to clearly demonstrate that rutabagastone can improve the quality of cryopreservation and in vitro maturation of frozen oocytes. By adding rutabagastone to the cryopreservation solution, thawing solution, and in vitro maturation solution, this invention reduces cryopreservation damage to oocytes, resulting in increased survival rate after thawing, improved mitochondrial function, reduced damage to the oocyte spindle and zona pellucida, reduced oxidative stress levels, and ultimately improved subsequent developmental capacity of frozen oocytes.

The invention relates to a mitochondrial delivery system and a preparation method and application thereof. According to the mitochondrial delivery system, safe and efficient cell entry of active mitochondria can be achieved, the mitochondrial delivery system has appropriate particle size and surface potential, toxicity to cells is avoided while the entrapment rate of the mitochondria is effectively improved and the electrochemical stability and biological activity of a mitochondrial membrane are guaranteed, and the mitochondrial cell entry efficiency is remarkably improved. The mitochondrial delivery system not only can transplant active mitochondria into common cells to repair cell damage, but also can transplant the active mitochondria into egg cells to play a role across a zona pellucida and a plasma membrane barrier. The mitochondrial delivery system can be used for treating mitochondrial diseases related to mitochondrial dysfunction, especially infertility caused by mitochondrial dysfunction of egg cells.

The application discloses a kind of high efficiency gentle mammal blastocyst single cell dissociation method.The method includes the following steps: (1) zona pellucida removal: the blastocyst is placed in calcium magnesium table type liquid, and zona pellucida is completely removed by dissolving and physical blowing operation;(2) enzyme digestion incubation: the blastocyst after step (1) is handled and is transferred to 35-37 ℃ pre-equilibrated complex dissociation solution and incubated for 15-30 minutes, during which intermittent blowing is carried out, until the blastocyst is completely dissociated into single cell;(3) washing: the single cell obtained by dissociation is washed using dPBS, and the residual dissociation solution is removed.The method of the application has high efficiency, high activity and simplified operation.The dissociation process is simplified from 3-4 steps to two steps (zona pellucida dissolution+enzyme digestion incubation), the total time is shortened from more than 90 minutes to less than 30 minutes, the dissociation efficiency is significantly improved;Cell viability is increased from about 70% to more than 90%, the cell junction is gently lysed, and cell damage is reduced.

The present application belongs to the technical field of pharmaceutical preparations, and specifically relates to the new use of Tongren Wuji Baifeng Pills and a pharmaceutical composition in the preparation of a drug for treating premature ovarian failure, a pharmaceutical preparation comprising Tongren Wuji Baifeng Pills, a method for preparing same, and a method for preparing a drug for treating premature ovarian failure. In the present application, a rat model of chemotherapy-induced premature ovarian failure via intraperitoneal injection of cisplatin and a mouse model of immune-mediated premature ovarian failure via multi-point subcutaneous injection of mouse zona pellucida peptide 3 (pZP3) are established. By means of observing the effects of Tongren Wuji Baifeng Pills on the estrous cycles, organ coefficients, serum sex hormone levels, and ovarian histopathological changes in animals in both models, the therapeutic effect of Tongren Wuji Baifeng Pills on premature ovarian failure is verified and clarified. This expands the new clinical use of the drug and provides a new therapeutic drug and means for treating premature ovarian failure, thereby offering technical support for clinical precision medication.

Provided herein are methods for improving the efficiency of in vitro fertilization of animals. The method comprises (a) obtaining an oocyte from the animal, wherein the oocyte comprises an oocyte body and a zona pellucida; (b) obtaining sperm from the animal; (c) maturing the oocytes in a maturing medium; (d) removing cumulus cells from the oocytes; (e) drilling in the zona pellucida of the oocyte with a laser; (f) contacting the oocyte with the sperm; (g) co-incubating the oocytes with sperms, whereby the incubation enables in vitro fertilization of the oocytes and sperms to produce animal embryos, and whereby drilling is performed in the zona pellucida of the oocytes by laser to improve the efficiency of in vitro fertilization of the production of animal embryos.

A method for automated ICSI includes receiving at least one droplet containing an egg in a dish placed on a stage. The method includes using an artificial intelligence / machine learning system (AI / ML system) and an imaging system to detect a zona pellucida. The imaging system includes a microscopy system, a camera system, and a lighting system. The method includes holding the egg using a robotic microtool and lowering a robotic pipettor into the droplet. The method includes using the AI / ML system and imaging system to determine an area at which to hold the egg and positioning the robotic microtool to that area. The method includes using the AI / ML system and imaging system to instruct the robotic microtool to apply negative pressure to hold the egg to the robotic pipettor. The method includes using the AI / ML system and imaging system to determine a target location where zona ablation should be performed.

The application relates to an image recognition-based method for positioning an oocyte polar body in an intracytoplasmic sperm injection, obtaining a microscopic digital image of an oocyte; determining an inner edge of a zona pellucida, an outer edge of a cytoplasm and a peripheral gap region located between the two according to the microscopic digital image; extracting a candidate region in the peripheral gap region; respectively judging whether each candidate region is located in the peripheral gap region, is not connected with the cytoplasm region and is not connected with the zona pellucida outer side region; determining a valid candidate region which is located in the peripheral gap region, is not connected with the cytoplasm region and is not connected with the zona pellucida outer side region; determining a target polar body region according to the valid candidate region; determining a polar body center position and polar body orientation information according to the target polar body region, and outputting a polar body positioning result. The method can be implemented based on a microscopic digital image, and is favorable for improving the stability, consistency and automation application value of the polar body positioning result.

The invention relates to the technical field of biological medicines, in particular to a construction method of Gjb2 homozygous mutation deafness mice. Injecting the Gjb2 homozygous mutant mouse embryonic stem cells into a diploid mouse embryo through zona pellucida to prepare a reconstructed embryo; and transferring the reconstructed embryo to a pseudo-pregnant female mouse, so that the postpartum generation mouse is the Gjb2 homozygous mutation deafness mouse. By adopting the construction method disclosed by the invention, the survival ratio of the mouse with homozygous mutation deafness is high, a large number of important mouse models can be provided for follow-up disease treatment exploration, and an important foundation is laid for efficient establishment of mouse models with other different mutations of GJB2.

The present invention provides a method for improving the efficiency of in vitro fertilization in animals. The method comprises: (a) obtaining oocytes from the animal, wherein the oocytes include the oocyte body and the zona pellucida; (b) obtaining sperm from the animal; (c) maturing the oocytes in a maturation medium; (d) removing cumulus cells from the oocytes; (e) creating holes in the zona pellucida of the oocytes with a laser; (f) contacting the oocytes with the sperm; and (g) incubating the oocytes with the sperm, wherein the incubation enables in vitro fertilization of the oocytes and sperm, and an animal embryo is produced; and the efficiency of in vitro fertilization for producing an animal embryo is improved by creating holes in the zona pellucida of the oocytes with a laser. In certain embodiments, the cumulus cells are removed from the oocytes by treatment with hyaluronidase, pipetting, and / or vortexing. In one embodiment, the oocytes are placed on a microscope stage prior to step (e).

Embodiments described herein use deep learning to automate measurement of key morphokinetic embryo features associated with viability and quality, in particular those relevant for clinical in-vitro fertilization (IVF). Systems and methods may, for example, acquire one or more digital images of one or more embryos; select one or more embryos in each digital image; and for each selected embryo, (i) computationally characterize the zona pellucida, detect the degree of fragmentation in the embryo, and for each embryo with a low fragmentation score, computationally classify the embryo's developmental stage based on whether cells constituting the embryo exceed a threshold number (e.g., nine). For an embryo consisting of a single cell, pronuclei may be detected and counted. Based on these measurements, a viability score may be assigned to the embryo.