9 results about "Maturation oocyte" patented technology

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 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.

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 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.

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.