23 results about "Maturation oocyte" patented technology

The invention discloses a pharmaceutical composition for delaying oocyte senescence and application thereof, and relates to the technical field of biological medicine, and the pharmaceutical composition comprises alpha-ketoglutaric acid, a bicarbonate buffer solution, an energy metabolism substrate, polyvinyl alcohol, hormone, a growth factor and insulin-transferrin-selenium. The pharmaceutical composition for delaying oocyte aging disclosed by the invention is beneficial to improving the maturation rate of oocytes, reducing the cytoplasm fragmentation rate and the cleavage fragmentation rate, and improving the cleavage rate and the blastocyst rate; according to the invention, the content of beta-galactosidase is reduced, the function of mitochondria in the aging process after ovulation of the mature oocyte is improved, the membrane potential of the mitochondria in the aging process after ovulation of the mature oocyte is stabilized, the activity of the mitochondria is improved, and the normalization of RNA methylation modification in the aging process after ovulation of the mature oocyte is promoted.

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 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.

The invention belongs to the technical field of embryo in-vitro culture, and discloses a method for in-vitro production of high-quality embryos by using frozen boar semen. In the in-vitro production process of the pig embryo, the fertilization rate and the quality of the in-vitro cultured embryo are improved by using the frozen semen treated by the antifreeze protein NT5C1B and the mature oocyte through in-vitro fertilization. Wherein the selected protein can reduce the generation of ice crystallization in the freezing process of the semen, improve the quality and fertilization ability of the frozen semen, and provide a basis for high-quality development of embryos after in-vitro fertilization. The application provided by the invention is simple and convenient to operate and remarkable in effect, and does not generate any harmful influence on animals. The invention provides a powerful theoretical basis for in-vitro production of high-quality embryos.

The present invention relates to a composition for prevention, improvement, or treatment of poor ovarian responders caused by aging, comprising butylphthalide or an acceptable salt thereof as an active ingredient, for which a statistically significant increase in the number and quality of mature oocytes was confirmed in 38- to 40-week-old mice (40 years old or older in humans) fed butylphthalide. Not only does the number of mature oocytes produced in the ovaries decrease in women as a result of aging, but also healthy mature oocytes (MII oocytes) decrease as a result thereof, thereby gradually reducing the possibility of pregnancy, but the active ingredient of the present invention may increase the number and quality of mature oocytes and thus has the effect of increasing the chances of pregnancy for older women. In addition, the present invention may inhibit a decrease in the number of primordial follicles caused by aging and thus has the effect of enabling the inhibition of diminished ovarian reserve caused by aging.

This invention provides a method to improve age-related decline in ovarian function by supplementing quiescent oocytes within primordial follicles. [Solution] The present invention provides a method for inducing mature oocytes, comprising the steps of: preparing a single-cell suspension from the ovary of a mammal other than a human; separating the single-cell suspension into oocytes and ovarian somatic cells (supporting cells) within a primordial follicle using a flow cytometer; re-aggregating the oocytes and ovarian somatic cells within the separated primordial follicle; and maturing the re-aggregated primordial follicle in vitro.

The invention belongs to the technical field of nucleus visual identification, and discloses a method for identifying and predicting the nucleus position of an oocyte by using a computer image, which comprises the following steps of: dividing the oocyte into a mature oocyte and an immature oocyte, and acquiring an oocyte image; dividing the oocyte image into a mature oocyte image and an immature oocyte image; carrying out visual imaging operation on the nucleus of the oocyte, and meanwhile, carrying out image annotation on an oocyte photo; performing data amplification on images containing mature oocytes and cell nucleus positions, and dividing the images into a training set and a verification set; a computer vision framework is trained using the data set. According to the method disclosed by the invention, the cell nucleus position of the mammal oocyte is predicted by utilizing a computer visual identification technology, the enucleation accuracy is improved from 53.84% to 92.30%, and the accuracy of judging the cell nucleus position of the oocyte in a somatic cell nucleus transplantation process is improved.

The invention discloses a culture solution for promoting in-vitro maturation of bovine oocytes and application of the culture solution, and belongs to the technical field of assisted reproduction. The invention provides the culture solution which is composed of brand new components and is used for promoting in-vitro maturation of the bovine oocytes, support is provided for maturation of the oocytes from multiple aspects of carrier transportation, energy metabolism regulation, signal transduction, extracellular environment and the like, the in-vitro maturation quality of the bovine oocytes can be effectively promoted, and the survival rate of the bovine oocytes is increased. The quality and consistency of nucleus maturation and cytoplasm maturation of oocytes are obviously improved; the cultured mature oocytes are used for in-vitro fertilization, so that the fertilization proportion and the blastocyst hatching rate can be remarkably improved; the provided culture solution is appropriate in component number, clear in proportion, simple and easy to operate in preparation method, short in time consumption, good in repeatability and suitable for batch production, and has a good application prospect in maturation culture of bovine in-vitro oocytes.

The application relates to the field of super-low-temperature cryopreservation of mammalian oocytes, and particularly relates to application of miR-32 in improving the development capacity of pig immature oocytes after freezing. Pig immature oocytes are frozen and cultured in vitro, and mature oocytes which are cultured to expel the first polar body are incubated in transfection liquid containing a miR-32 nucleotide sequence for 2-6 hours; the pig immature oocytes are GV stage oocytes; and the miR-32 nucleotide sequence is shown in SEQ ID NO:1. It is found that miR-32 can be used as a key marker molecule of damage of vitrification oocytes, and by transfecting miR-32 mimics into the frozen oocytes, the early embryo development capacity of the oocytes can be improved, a new solution is provided for improving the maturation quality of the frozen oocytes, and various embryo biological technology researches are facilitated.

The application discloses a culture medium for improving in-vitro aging of oocytes after ovulation and application thereof. The culture medium is prepared by adding 0.1-30 muM of astragaloside IV into an oocyte culture solution. The culture medium is safe, non-toxic and free of side effects, can reduce the oocyte fragmentation rate, active oxygen level and abnormal spindle rate of the oocytes after 24 hours of in-vitro culture, can significantly improve the mitochondrial membrane potential, in-vitro sperm binding capacity, in-vitro fertilization pronucleus rate and cleavage rate of the mature oocytes after 24 hours of in-vitro culture, and can improve the quality of the late embryos. The application of the culture medium provides favorable support for assisted reproductive technology, and has a good application prospect.

The present invention relates to the field of biotechnology and provides a composition, culture medium, product, and method for promoting in vitro maturation of immature oocytes. The composition comprises KT5823 and gastrodin. The composition can reduce the levels of pro-inflammatory factors in oocytes, alleviating the toxic effects of inflammation and autophagy on oocytes. Furthermore, it can effectively reduce reactive oxygen species (ROS) levels and increase the antioxidant glutathione content, thereby alleviating oxidative stress damage to oocytes. Through these synergistic effects, the composition significantly reduces oocyte apoptosis, significantly improves the quality of in vitro oocyte maturation, and significantly increases the development rate after in vitro fertilization.

A system for predicting the number of mature oocytes in a subject, comprising: a data acquisition module, used for obtaining data of the age, basal anti-Mullerian hormone (AMH) level, basal follicle-stimulating hormone (FSH) level or basal antral follicle count (AFC), and dynamic change of an inhibin B level in an early follicular phase (ΔINHB / difference between inhibin B on the sixth day and inhibin B on the 2nd day of menstruation in an ovulation induction cycle) of the subject; and a mature oocyte number calculation module, used for calculating the data obtained in the data acquisition module, so as to calculate the number of retrieved mature oocytes (NROs) in the subject. According to the system and method of the present application, the dynamic change of the inhibin B level in the early follicular phase is used as an evaluation index to replace an AFC index having a plurality of defects in the prior art, so that a better effect of predicting the NROs is achieved. The system can be used for predicting the NROs in an ovulation induction process (such as the sixth day of menstruation in the ovulation induction cycle) according to a dynamic change index, thereby adjusting the dosage of an ovulation induction drug.

A method of providing a culture of oogonia stem cells comprising oogonia stem cells is provided. The method may further include culturing the oogonia stem cells with granulosa and theca cells to differentiate the oogonia stem cells into oocytes. In some embodiments, the culturing comprises including the oogonia stem cells in an in vitro follicle construct or a microcapsule comprising said granulosa and theca cells. Further described herein is a method of forming a bioengineered follicle construct capable of releasing a mature oocyte. An in vitro fertilization method using the mature oocyte is also provided.

The invention provides a culture solution for improving an oocyte in-vitro maturation defect caused by diabetes and application of BGP-15 or the culture solution, and particularly belongs to the technical field of oocyte culture. The invention provides an application of BGP-15 in preparation of a product for improving an oocyte in-vitro maturation defect caused by diabetes mellitus. The product further comprises an oocyte basal culture medium. BGP-15 is combined with an oocyte basic culture medium, so that the first polar body discharge rate of in-vitro mature oocytes of diabetic mice can be remarkably increased, the oxidative stress level of the oocytes is reduced, the mitochondrial membrane potential of the oocytes is recovered, and the survival rate of the oocytes is increased. The in-vitro maturation rate of the diabetic mouse oocytes is increased, the quality of the in-vitro mature oocytes of the diabetic mouse is improved, and the method has important guiding significance in treatment of diabetic female infertility.

The invention relates to the technical field of biology, and provides a composition, a culture solution, a product and a method for promoting in-vitro maturation of immature oocytes. The composition is prepared from KT5823 (KT5823) and Gastrodin (Gastrodin). The composition can reduce the proinflammatory factor level of oocytes and relieve the toxic effect of inflammatory response and autophagy response on the oocytes. Meanwhile, the active oxygen level can be effectively reduced, the content of glutathione serving as an antioxidant substance is increased, and damage of oxidative stress to oocytes is relieved. Through the synergistic effects, the composition obviously reduces the apoptosis rate of oocytes, obviously improves the in-vitro maturation quality of the oocytes, and greatly improves the development rate after in-vitro fertilization.

The present invention relates to the field of in vitro maturation culture of oocytes, and particularly to an additive, a culture medium for promoting the quality of in vitro matured bovine oocytes and their applications; the additive for promoting the in vitro maturation of oocytes is NAD+, and the concentration of the additive NAD+ is 50 μmol / L. A culture medium for promoting the in vitro maturation of oocytes, the components of the culture medium are: M199 medium, 10 mmol / L Hepes, 10% FBS, 0.01 μg / mL FSH, 1 μg / mL LH, 0.01 μg / mL E2, 10-100 μmol / L NAD+. Adding NAD+ to the in vitro maturation culture medium of oocytes can effectively increase the number of mature oocytes and improve the quality of in vitro matured oocytes. By adding NAD+ to the in vitro maturation solution of oocytes, the in vitro maturation efficiency of oocytes can be improved, the existing oocyte maturation culture system can be improved, and then it can be applied to technical fields such as in vitro fertilization, parthenogenetic activation, somatic cell cloning, etc., so as to improve its production and application value.

The invention discloses a culture solution and a method for improving in-vitro maturation and quality of remaining GV stage ova after ICSI, and relates to the field of assisted reproduction. The invention provides a culture solution for improving in-vitro maturation and quality of remaining GV stage ova after ICSI, the culture solution is a BM-CoE finished product culture solution composed of 50 [mu] M of coenzyme Q10, 10 ng / ml of epidermal growth factors and a BM basic culture solution, and the BM basic culture solution is composed of a finished product blastocyst culture solution 1029, a 10% serum protein substitute, 29 [mu] M of sodium pyruvate and 1.5 IU / ml of human menopausal gonadotropin. By jointly adding the coenzyme Q10 and the epidermal growth factor with certain concentration into the BM basic culture solution, compared with a commercially available finished product oocyte in-vitro maturation culture solution, the culture solution disclosed by the invention not only can obtain higher oocyte maturation rate and better mature oocyte quality, but also is beneficial to improving the clinical oocyte utilization rate and increasing the number of available embryos of a patient.

The invention relates to the technical field of assisted reproduction, and discloses an in-vitro maturation culture system and method for immature oocytes of elderly women based on NMN supplement. The culture system is an in vitro maturing (IVM) culture solution containing 10 [mu] M of NMN (Nitrogen Morphocyte Nitride), and the culture system is an in vitro maturing (IVM) culture solution containing 10 [mu] M of NMN; the culture method comprises the following steps: preparing an IVM culture solution; culturing in vitro; carrying out in-vitro fertilization; and early embryo development observation. According to the application, NAD < + > is improved through targeted supplement of NMN, the mitochondrial function is further repaired, a brand new path for cooperatively regulating meiosis is achieved, and the problem that in the prior art, only oxidative stress is relieved, but the energy supply defect cannot be solved is solved. Experimental data show that the IVM system added with the NMN can remarkably improve the blastocyst formation rate of oocytes of elderly women, the mitochondrial function is recovered, and the NMN-containing IVM system has a remarkable clinical transformation value.

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 invention provides application of an AIM2 expression inhibitor in preparation of a medicine for treating ovulation disorder of polycystic ovarian syndrome. Experiments show that AIM2 is highly expressed in ovaries and / or granulosa cells of a polycystic ovarian syndrome ovulation disorder patient and an animal model, and further show that by adding A151 to inhibit expression of AIM2, ovarian polycystic change can be reversed, the number of sinus follicles and atresia follicles is reduced, the number of corpus luteum, the number of oocytes and the rate of mature oocytes are increased, and the effect of inhibiting ovulation disorder of the polycystic ovarian syndrome is achieved. Therefore, the expression inhibitor of the AIM2 can be used for preparing a medicine for treating ovulation disorder of the polycystic ovarian syndrome.

A method of providing a culture of oogonia stem cells comprising oogonia stem cells is provided. The method may further include culturing the oogonia stem cells with granulosa and theca cells to differentiate the oogonia stem cells into oocytes. In some embodiments, the culturing comprises including the oogonia stem cells in an in vitro follicle construct or a microcapsule comprising said granulosa and theca cells. Further described herein is a method of forming a bioengineered follicle construct capable of releasing a mature oocyte. An in vitro fertilization method using the mature oocyte is also provided.

The present disclosure relates to methods for producing mature oocytes comprising transplanting ovarian tissue or any fragment thereof of a human female into at least one recipient site of an immunodeficient rabbit, and collecting mature oocytes from said transplanted ovarian tissue. In addition, the methods comprises providing conditions for follicles development and oocytes maturation in said rabbit.

The invention provides application of an expression inhibitor of HAS2 in preparation of a medicine for treating ovulation disorder of polycystic ovarian syndrome. Experiments show that HAS2 is highly expressed in ovaries and / or granular cells of patients suffering from ovulation disorder of polycystic ovarian syndrome and animal models, and further show that ovarian polycystic change can be reversed by adding hydroxymerocoumarin to inhibit expression of HAS2, the number of sinus follicles and atresia follicles is reduced, and ovarian ovulation disorder can be inhibited. The quantity of corpus luteum, the quantity of oocytes and the mature oocyte rate are increased, so that the expression inhibitor of the HAS2 can be used for preparing the medicine for treating the ovulation disorder of the polycystic ovarian syndrome.