31 results about "Mature 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 present invention relates to the field of biotechnology, and the present invention provides a composition, a culture medium, a product and a method for alleviating the aging of mature eggs. The composition comprises SB265610, MTK458 and 8-Br-cGMP. The composition can reduce the level of inflammatory factors and the toxic effect of inflammatory reactions on eggs. At the same time, it can increase the number and activity of mitochondria to provide sufficient energy guarantee for eggs. In addition, the composition can also effectively reduce the level of reactive oxygen species and the damage of oxidative stress to eggs. Through a series of combined effects, the composition significantly reduces the incidence of egg apoptosis and the rate of spindle abnormalities. By using the composition of the present invention, the aging rate of mature eggs is significantly alleviated, and the development rate is increased by nearly 20 times. The present invention provides a new strategy for animal embryo engineering and human assisted reproductive technology and has broad application prospects.

The present invention relates to a culture medium composition for in vitro maturation of oocytes, comprising piperine as an active ingredient, and an in vitro embryo production method using same. The culture medium composition for in vitro maturation of oocytes, comprising piperine as an active ingredient, of the present invention, can qualitatively improve oocytes by inhibiting reactive oxygen species, and in vitro embryo production efficiency can be increased using in vitro matured oocytes obtained from the medium of the present invention.

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.

The invention provides application of an RTL6 gene in improvement of an in-vitro embryo development rate, and relates to the technical field of mammal embryo engineering. According to the present invention, the RTL6 gene is constructed into the PcDNA3.1 (+)-RTL6 recombinant vector, and the pcDNA3.1 (+)-RTL6 recombinant vector is injected into the in vitro mature oocyte cytoplasm 4-5 h after the pig parthenogenetic activation, such that the embryonic development retardation phenomenon is effectively relieved, and the embryonic development is promoted;

The invention discloses a method for improving the normal fertilization rate of an in-vitro ICSI embryo of a dairy cow and reducing the abortion rate, which comprises the following steps: preparing an ICSI egg from X-sex controlled sperm and an oocyte which is cultured by a mature oocyte and is used as a donor by adopting a microinjection mode, the ICSI eggs are incubated in an oocyte maturation culture solution (OM) for 3-5 h and then transferred to ETH to be activated for 4-6 min so as to promote normal fertilization of the ICSI eggs, then embryo culture is conducted, and then sexual control embryos are obtained; the ETH refers to an oocyte maturation culture solution containing 6 to 8 percent of ethanol. The normal fertilization rate of the ICSI embryos produced by the method is remarkably improved, and the early abortion rate is remarkably reduced in inspection after transplantation.

The application belongs to the technical field of pest control, and discloses application of a transcription factor CG17328 in control of drosophila melanogaster. The application discloses application of CG17328 in regulation of insect reproductive capacity. The application firstly screens a transcription factor for regulating the 'dumping' process of nurse cells in oogenesis of drosophila melanogaster by using transcriptome sequencing, genetics and molecular biology and other related technologies and means. The transcription factor is a ZAD-C2H2 type zinc finger protein CG17328. CG17328 is mainly located in the nuclei of nurse cells and oocytes in the late stage of oogenesis. CG17328 can affect the oogenesis process by regulating the 'dumping' process of nurse cells. The loss of CG17328 expression leads to significant decrease of the number of mature oocytes generated by ovaries and the hatching rate of eggs, which indicates that the loss or inhibition of CG17328 expression can achieve the control effect on drosophila melanogaster.

The invention belongs to the technical field of biology, and particularly relates to a method for improving in-vitro mature oocyte telomere length and maturation quality and application of the in-vitro mature oocyte telomere length and the maturation quality. And daphnetin. According to the preparation disclosed by the invention, at least one of the Sema3 type protein and the daphnetin is added, so that the telomere length of an oocyte and an in-vitro embryo is effectively maintained, repaired or prolonged, the maturation quality of the in-vitro oocyte and the development potential and development efficiency of the in-vitro embryo are further remarkably improved, and further, the development potential and development efficiency of the in-vitro embryo are further improved. The application provides a more effective new strategy for in-vitro embryo production of livestock, animal genetic improvement, assisted reproduction technology and the like, and the application prospect is wide.

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.

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 discloses a sheep in-vitro fertilization culture method based on a serum substitute, and belongs to the technical field of biology. The method comprises the following steps that mature oocytes of sheep are cultured through in-vitro fertilization liquid, the in-vitro fertilization liquid does not contain oestrus sheep serum or fetal calf serum, and the in-vitro fertilization liquid contains serum substitutes. Experiments show that 10% XRS replaces 10% FBS, and an ideal oocyte maturation rate can be obtained; xRS replaces OSS, the cleavage rate can be increased, and therefore the XRS replacement scheme is feasible in the sheep germ cell in-vitro culture technology.

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.

A method for inducing immature oocytes includes introducing four genes consisting of FIGLA, NOBOX, LHX8 and TBPL2, or transcripts or expressed proteins thereof, into at least one type of cell selected from the group consisting of pluripotent stem cells, epiblast-like cells and primordial germ cells. A method for producing mature oocytes includes introducing four genes consisting of FIGLA, NOBOX, LHX8 and TBPL2, or transcripts or expressed proteins thereof, into at least one type of cell selected from the group consisting of pluripotent stem cells, epiblast-like cells and primordial germ cells, and co-culturing the cell obtained after the introduction and ovarian somatic cells.

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 discloses a method for improving chemical activation efficiency of oocytes. The method comprises the following steps: designing a combination mode of ionomycin treatment concentration, ionomycin treatment time, 6-DMAP treatment concentration and 6-DMAP treatment time through an orthogonal test; (2) placing the mature oocytes in a DMEM / F12 culture medium containing ionomycin and a PZM-3 culture medium containing 6-DMAP according to a specific combination mode for parthenogenetic activation treatment; the treatment concentration of ionomycin is 6-12 [mu] M, and the treatment time is 3-6 min; the treatment concentration of the 6-DMAP is 1.8 to 2.1 mM, and the treatment time is 4 to 7 hours. According to the method, the blastocyst rate reaches 66.53 + / -1.71% by designing an orthogonal test, systematically optimizing a specific chemical activation combination mode of ionomycin and 6-DMAP and based on parameter combination optimized by the orthogonal test, and compared with an existing chemical activation scheme, the absolute value of the blastocyst formation rate is increased by more than 36%.

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.

The invention discloses an application of gene expression regulation in rescue of developmental capacity of aged oocytes after ovulation, and relates to the technical field of biological medicines, which comprises regulation of YAP1 gene knock-down and GOT2 gene overexpression of the aged oocytes after ovulation. The developmental phenotypes (maturation rate, cleavage rate, blastocyst rate, cytoplasm fragmentation rate and cleavage fragmentation rate) of the cultured aged oocytes after ovulation in combination with YAP1 knock-down and GOT2 overexpression tend to be close to normal mature oocytes which are not subjected to aging treatment after ovulation, and the developmental ability of the aged oocytes after ovulation can be saved.

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 relates to the technical field of animal breeding biology, and particularly discloses a method for improving the development efficiency of in-vitro embryos of young cattle and sheep, which comprises the following steps: collecting oocytes from an in-vitro ovary, washing by using a conventional in-vitro maturation solution added with enoxone, and drying to obtain the in-vitro embryos of the young cattle and sheep. Carrying out pretreatment culture for 2-4 hours in a conventional in-vitro maturation culture solution added with enoxone, and then transferring into the conventional in-vitro maturation culture solution to continuously culture for 20-26 hours; and carrying out in-vitro fertilization on the mature oocytes to obtain fertilized eggs, and carrying out in-vitro culture for 8-10 days. According to the method provided by the invention, the cleavage rate and blastocyst development rate of oocytes of calves and lambs after in-vitro fertilization are improved by adding Enoxilone into an in-vitro maturation culture solution.