13 results about "Cumulus Cell" patented technology

The present disclosure generally relates to micromanipulation devices and methods for the manipulation of mammalian sub-cellular structures. Uses include intra cytoplasmic sperm injection procedures, embryo biopsy procedures, trophectoderm biopsy procedures, mitochondrial transfer procedures, pronuclear transfer procedures, maternal spindle transfer procedures, cumulus cell biopsy procedures and other sub-cellular manipulation procedures. Devices comprise a cellular manipulation dish further comprising multiple sub- cellular manipulation units having a microwell and a cell deposit well wherein the cell deposit opening is disposed adjacent to the microwell opening.

The invention relates to the technical field of biomedical treatment, and discloses a cumulus cell stripping quality assessment method and device.The method comprises the steps that on different focusing layers, two focusing images of an ovum to be assessed after cumulus cells are stripped are collected, and the positions of all cumulus cells and the total number of the cumulus cells are recognized; summing the total number of cumulus cells in the two focused images to obtain an initial number; intercepting each cumulus cell in the two focused images, and pairing each cumulus cell in the first focused image and each cumulus cell in the second focused image to obtain a plurality of cumulus cell pairs; identifying the logarithm of the cumulus cell pairs of which the two cumulus cells are the same cumulus cell, and based on the position of each cumulus cell, rejecting the cumulus cell pairs of which the position difference is greater than a preset distance to obtain a possible cell logarithm; and calculating the difference between the initial number and the possible cell logarithm to obtain the number of cumulus cells around the to-be-evaluated ovum, and obtaining the cumulus cell stripping quality of the to-be-evaluated ovum.

Microfluidic systems are configured to process liquid samples that include cumulus-oocyte complexes (COCs), such as raw follicular fluid, in an automated and continuous manner to produce oocytes separate or “denuded” from surrounding cumulus cells. The systems include a substrate and a channel that can have two or more sub-channels. Each channel includes an inlet, an outlet, and one or more stages arranged in series. Each of the stages includes one or more expansion units and one or more constriction units. At least one stage includes constriction units having jagged internal surfaces, e.g., teeth, that help remove the cumulus cells from the COCs.

The invention belongs to the technical field of medical biology, and particularly relates to application of porcine cumulus cell miR-31 in improvement of maturation quality of oocytes, and the nucleotide sequence of the porcine cumulus cell miR-31 is shown as SEQ ID NO.1. The invention also relates to a preparation method of the porcine cumulus cell miR-31. The miR-31 can improve the cumulus expansion level, improve the oocyte quality, enhance the mitochondrial function and reduce oxidative stress, target MAPK8 to participate in a JNK channel, inhibit the apoptosis level of the oocyte and remarkably improve the in-vitro oocyte quality. The miR-31 can be used as a potential biomarker for maturation of oocytes. The results provide a theoretical basis for further discussing the effect of miRNA participating in proliferation and differentiation of animal reproduction related cells and the action mechanism of miRNA on follicle development and oocyte maturation.

The invention discloses application of a compound shown in a formula I in promoting in-vitro cultured oocyte development or in-vitro embryo development, and in the formula I, R1 is selected from halogenated alkyl; r2 is selected from H, halogen or C1-6 alkyl; n is selected from 0, 1, 2 or 3. According to the application, the expansion area of cumulus cells in the cumulus-oocyte complex can be increased; the oxidative damage of the cumulus-oocyte complex is relieved; the first polar body discharge rate of the oocytes is improved; the abnormal distribution rate of mitochondria in oocytes is reduced; the membrane potential of mitochondria in the oocyte is improved; and the cleavage rate and blastocyst formation rate of fertilized eggs can be improved, and the apoptosis rate of oocytes can be reduced.

Cumulus cell (CC) gene expression is explored as an additional method of morphological scoring to select an embryo with the highest chance of pregnancy. The present invention relates to a novel method for identifying biomarker genes for assessing the ability of mammalian oocytes to produce viable pregnancy after fertilization based on the use of live birth and embryonic development as endpoint criteria for the oocytes for exon level analysis of potential biomarker genes. The invention further provides biomarker genes of CC expression thus identified, as well as prognostic models based on biomarker genes identified using the methods of the invention.

The invention belongs to the field of cell culture, and particularly discloses application of dominant follicle exosomes in oocyte in-vitro maturation culture, after the dominant follicle exosomes with the concentration of 10 ng / mL-200 ng / mL are added into a porcine oocyte in-vitro maturation culture solution for porcine oocyte in-vitro maturation culture, the first polar body discharge rate is remarkably increased, and the first polar body discharge rate is remarkably increased. Therefore, the dominant follicle exosome can effectively improve the in-vitro maturity of the porcine oocytes; the dominant follicle exosomes with different concentrations are added, so that the diffusion area of cumulus cells is remarkably increased; after the dominant follicle exosome is added, the ROS fluorescence intensity is obviously reduced, and the GSH fluorescence intensity is obviously increased; by adding the dominant exosome, the remodeling of the mitochondrial structure and function in the oocyte is accelerated; adding of the dominant exosome promotes functional remodeling of the mitochondrial complex I to increase ATP generation and reduce oxidative stress so as to promote nucleoplasm co-maturation of oocytes. The subsequent embryonic development can be promoted.

The invention provides application of a cumulus cell mTOR-S6 protein marker in non-invasive prediction of embryo quality and pregnancy outcome, and relates to the technical field of bioengineering. According to the present invention, the Westernblot technology is adopted to quantitatively detect the levels of the mTOR signal channel key proteins such as p-mTOR, mTOR, p-S6 and S6 in cumulus cells; meanwhile, the system collects IVF related clinical indexes of the patient, correlation analysis is carried out on the clinical indexes and the relative levels of the four key proteins through a statistical method, and it is found that the IVF related indexes of the patient are evaluated with the median value of the relative expression values of the four proteins as a threshold value. Whether the ovarian reserve function, the embryonic development quality and the pregnancy outcome are good or not can be judged. The invention not only provides an innovative auxiliary evaluation tool for the field of reproductive medicine, but also provides a scientific basis for formulating an individualized IVF treatment scheme.

A method for automated, artificial-intelligence-based denudation includes scanning, robotically, a dish containing a cumulus-oocyte-complex (COC) using an imaging system and an artificial intelligence / machine learning system (AI / ML system) to create an image object. The imaging system includes a microscopy system, a camera system, and a lighting system. The dish includes an enzyme to remove cumulus cells from an oocyte among the COC. The method includes identifying the COC within the image object, and a location, based on comparing the image object to a threshold using the AI / ML system. The threshold is an optical pattern with a probability of corresponding to a COC mass. The method includes instructing a robotic pipettor to collect the COC at the location. The method includes iteratively collecting the COC within the robotic pipettor and expelling the COC to return it to the dish until the AI / ML system confirms sufficient removal of cumulus and corona cells.

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.

The invention discloses an efficient immature oocyte microinjection method which comprises the following steps: sucking an oocyte washing liquid by using a pipettor, adding the oocyte washing liquid into a culture dish until the oocyte washing liquid submerges the bottom of the culture dish, then adding immature oocytes, aligning a cumulus oocyte complex in the culture dish by using the pipettor, and gently blowing and beating to remove part of cumulus, the method comprises the following steps: sucking one oocyte by using a common oocyte transfer needle, placing the oocyte in a T10 liquid drop, pressing a part of a residual cumulus cell by using a small-caliber oocyte transfer needle, gently and repeatedly sucking and blowing until part of oocyte zona pellucida is exposed, transferring the oocyte zona pellucida into another T10 liquid drop, and cleaning; the exposed zona pellucida part of the oocyte is aligned with the microinjection needle, and when the needle tip of the microinjection needle is observed to be tangent to the spherical surface of the oocyte under a microscope, the needle tip is inserted into cytoplasm from the exposed zona pellucida part to inject siRNA. According to the invention, efficient injection of immature oocytes can be realized, and meanwhile, the influence on maturation of the oocytes is smaller.

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).

The invention discloses a method for predicting OPU-IVF blastocyst development and embryo production performance by identifying a multi-gene expression pattern in an oocyte and a cumulus cell, which comprises the following steps: detecting expression patterns of mitochondrial functional genes ISCU and IGF2BP3 in the oocyte and an oocyte-cumulus cell interaction gene CTNND1; the method further comprises the step of detecting expression modes of an oxidative phosphorylation and metabolism regulation pathway SOX4, a gene KLF12 participating in cell cycle control and apoptosis regulation, an oxidative phosphorylation and metabolism regulation pathway OGT and an oocyte marker FST in cumulus cells. The expression modes of the genes can predict the embryonic development potential of oocytes after in-vitro maturation and in-vitro fertilization, important genetic information prediction is provided for predicting the reproductive performance of livestock, especially applying the in-vitro fertilization technology to embryo production of large livestock, and the breeding and breeding process of high-value livestock populations is promoted.