32 results about "Immature oocyte" patented technology

The invention discloses an in-vitro maturation culture method for an oocyte mouse and a method for establishing a parthenogenetic embryonic stem cell line. In the in-vitro maturation culture method, an immature oocyte is cultured in a basic culture solution in which the HCG (human chorionic gonadotropin) and PMSG (pregnant mare serum gonadotropin) are added, and the maturation rate of the oocyte can reach more than 83 percent. In the method for establishing a parthenogenetic embryonic stem cell line, the parthenogenetic activation culture development is carried out on the mouse oocyte after in-vitro maturation culture to obtain the embryonic stem cell line; a hepatocyte obtained by the method has no immunogenicity, is simultaneously equivalent to a stem cell derived from a fertilized embryo on totipotency and has guiding significance for utilizing in-vitro maturation of the immature oocyte of a human and separating parthenogenetic embryonic stem cells. Aiming at the current conditionsof human ovum donation shortage, great discarding of clinical immature oocytes by an assisted reproductive technology, and the like, the two methods are combined to lay the foundation for developing and utilizing the in-vitro maturation culture of the immature oocyte of people and further researching the parthenogenetic embryo by utilizing an in-vitro mature ovum and separating the embryonic stemcells.

The extracorporeal culture and mature method of immature oocyte from ovary tissue block includes selecting marrow mesenchyme stem cell MSC of female mouse as the feeder cell, compounding MSC culture liquid and agar and covering the culture rack, adding HCG in 3 IU / ml, and adhering the ovary tissue block on the rack to culture. In the co-culture system, estradiol, progestin and luteotopin contents are increased obviously, and after culturing, there is mature oocyte discharged. The method is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

The present invention relates to the technical field of assisted reproduction and discloses an in-vitro culture solution of immature oocytes. The in-vitro culture solution comprises a basic culture solution and a composition. The basic culture solution comprises a solvent and a solute, the solvent comprises water, and the solute comprises vitamins, energy substances, inorganic salts and amino acids; and the composition comprises 0.01-1 mg / L of butyrolactone-I, 0.001-2 mg / L of estradiol, 0.001-1 IU / L of follicle-stimulating hormone, 0.001-1 IU / L of luteinizing hormone, 0.001-1 IU / L of epidermalgrowth factor, 0.001-1 mg / L of insulin-like growth factor and 0.001-1 IU / L of human chorionic gonadotropin. The present invention also discloses a preparation method and an application of the in-vitro culture solution. In addition to adding basic substances, by adding the energy substances, composition and vitamins, nucleus and cytoplasm of the immature oocytes are synchronously mature and besides, a fertilization rate and a high-quality embryo rate are improved.

An assembly is disclosed that is to be used for the culturing of specimens such as embryos and gametes for use in in vitro fertilization. The assembly includes an annular ring which is affixed to the stage of an optical viewing instrument, such as a microscope. The viewing instrument is focused on a point which lies inside of the ring at a predetermined distance from the center of the ring. Circular specimen dishes having a plurality of specimen wells in which the specimens in question are cultured or grown is removably positioned inside of the ring. The specimen wells have bottom walls which are configured so as to ensure that the specimens in the wells will gravitate to the same predetermined position in each of the wells. That position coincides with the focus point of the viewing instrument. When the specimen culturing dishes are placed inside of the ring, they can be rotated inside of the ring to bring the specimens in each well sequentially into the focus point of the viewing instrument whereby the specimens in each dish can be quickly and accurately monitored for growth and development. Other devices are disclosed which enable the rotation of the dish and proper alignment of the specimens in each well with the focus point of the viewing instrument. The assembly can be used for a wide range of specimen culturing. Typical specimens include animal and human cells, tissues, stem cells, embryos, oocytes, immature oocytes, sperm precursor cells, embryonic cells, blastocysts, and spermatozoa.

The invention provides an application of enhancing antioxidation of an oocyte cultured in vitro by a Peroxiredoxin 4 protein. The maturing rate of the immature oocyte cultured in vitro can be improvedby improving the resistance of particle cells in oxidative stress and improving the antioxidation of the oocyte. Aging of the oocyte is also delayed through an antioxidant stress mechanism in the cell, and the fertility rate and the clinical pregnancy rate of the oocyte in an assisted breeding process are improved.

The invention provides methods for in vitro maturation of immature human oocytes. In one aspect, the invention provides a method for in vitro maturation of immature human oocytes, comprising the steps of: (a) inducing in a female human subject an increase in endogenous luteinizing hormone levels, said subject having not undergone an ovarian stimulation protocol prior to said inducing step; (b) obtaining from said subject an immature oocyte; and (c) culturing said oocyte until maturity.

The invention discloses mature optimized human oocyte cytoplasm fluid. The invention develops the mature optimized human oocyte cytoplasm fluid by adding some important hormones, energy substances and efficient antioxidant melatonin on the basis of low-sugar TCM199 tissue culture solution and patient autoserum. The invention develops a culture solution which is suitable for mature optimization of the oocyte cytoplasm in IVF-ET period and also is suitable for reutilization after in vitro maturity of the immature oocyte in the period. The mature optimized human oocyte cytoplasm fluid is capable of improving the maturity degree of the cytoplasm of the matured oocyte. A clinical result proves that the high-quality embryo rate of the oocyte is greatly increased, the development potential of the immature oocyte is increased and some immature oocytes in the IVF-ET period have a reutilization value.

The extracorporeal culture and mature method of immature oocyte from ovary tissue block includes selecting marrow mesenchyme stem cell MSC of female mouse as the feeder cell, compounding MSC culture liquid and agar and covering the culture rack, adding HCG in 3 IU / ml, and adhering the ovary tissue block on the rack to culture. In the co-culture system, estradiol, progestin and luteotopin contentsare increased obviously, and after culturing, there is mature oocyte discharged. The method is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

The invention proposes a genetic modification method. The method comprises: in situ microinjection of genetic modification reagents to immature oocytes in model animals. Using the genetic modification method according to the embodiment of the present invention, the time of microinjection is advanced to the period when the eggs are not yet mature, and the method of direct injection in the mother is adopted. After the injected oocytes mature naturally in the mother, they are injected After excretion and fertilization, mutants with edited genomes in almost all cells can be obtained in the F0 generation, and the phenotypes consistent with the homozygous mutants can be seen in the injected contemporary embryos, as well as a stable expression of the transgene map. With technology, the time to obtain mutants is shortened to 2-3 months, and large-scale screening work is not required.

The present invention is feeder cell for extracorporeal culture of immature oocyte and features that marrow mesenchyme stem cell MSC of female mouse is used as the feeder cell. The feeder cell is easy to obtain, can secrete several kinds of cell factors and growth factors, promote over 90 % of the immature oocyte to become mature and promote the growth, ovulation and maturing of the ovarian follicles in the ovary tissue block under extracorporeal culture. The present invention is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

The invention relates to an immature oocyte in vitro maturation promoting culture medium and application thereof. The immature oocyte in vitro maturation promoting culture medium is screened out through research on effects of different additives on immature oocytes and prepared by adding 20-100 mu M of BAPTA-AM in normal oocyte in vitro maturation culture solution. The immature oocyte in vitro maturation promoting culture medium is safe and free to side and toxic effects, enables meiosis of the immature oocytes, particularly those in level 3 cumulus-oocyte complexes, improves the in vitro development of the oocytes, promotes maturation, improves the cleavage rate and the blastula development rate after the mature oocytes are parthenogenetically-activated and further improves embryo quality. Application of the immature oocyte in vitro maturation promoting culture medium provides favorable support for the technology of in vitro propagation of animal embryos.

The invention relates to the technical field of artificial reproduction of fish, in particular to a method for improving the efficiency of artificial reproduction of long thin loach. The specific steps are as follows. Firstly, the broodstock are carefully screened and temporarily kept in polyvinyl chloride cages, and then the broodstock are classified. According to different developmental levels, different oxytocin needle intervals and drug dosages are selected, which is beneficial to the protection of the broodstock. With the synchronous maturation of oocytes, the proportion of immature eggs in the induced oocytes decreases from 10% to 3%, and then the induced oocytes are treated to mix the sperm and eggs evenly, and finally the broodstock is nursed to minimize the number of spawning broodstock. The injury and the death of postpartum broodstock have protected the limited long thin loach resources. The mortality rate of postpartum broodstock in traditional methods is as high as 70% to 100%, and the mortality rate is 10% under the technical conditions.

The invention relates to a novel bovine oocyte in vitro maturation (IVM) culture solution, belonging to the field of embryo biotechnolory. In the invention, the routine IVM culture solution is added with amniotic epithelia or the supernatant of the culture solution thereof to establish a novel bovine immature oocyte IVM culture system, so as to improve the quality and maturation rate of the IVM bovine oocyte, thus solving the problem of low fertilization rate and low formation rate of blastula in the existing culture method.

The method of obtaining immature oocyte from ovary tissue through separating or extracting for extracorporeal culture includes selecting marrow mesenchyme stem cell MSC of female mouse as the feeder cell or culturing the immature oocyte in the MSC culture liquid. After culturing, the co-culture system has obviously increased estradiol, progestin and luteotopin contents, and over 90 % of the immature oocyte become mature. The method is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

The invention discloses an in-vitro maturation culture method for an oocyte mouse and a method for establishing a parthenogenetic embryonic stem cell line. In the in-vitro maturation culture method, an immature oocyte is cultured in a basic culture solution in which the HCG (human chorionic gonadotropin) and PMSG (pregnant mare serum gonadotropin) are added, and the maturation rate of the oocyte can reach more than 83 percent. In the method for establishing a parthenogenetic embryonic stem cell line, the parthenogenetic activation culture development is carried out on the mouse oocyte after in-vitro maturation culture to obtain the embryonic stem cell line; a hepatocyte obtained by the method has no immunogenicity, is simultaneously equivalent to a stem cell derived from a fertilized embryo on totipotency and has guiding significance for utilizing in-vitro maturation of the immature oocyte of a human and separating parthenogenetic embryonic stem cells. Aiming at the current conditions of human ovum donation shortage, great discarding of clinical immature oocytes by an assisted reproductive technology, and the like, the two methods are combined to lay the foundation for developing and utilizing the in-vitro maturation culture of the immature oocyte of people and further researching the parthenogenetic embryo by utilizing an in-vitro mature ovum and separating the embryonic stem cells.

The present invention relates to cell culture, and specifically discloses a culture scheme for immature oocytes, which can promote the in vitro maturation of immature oocytes, specifically comprising follicular granulosa cells and a culture medium for cultivating the follicular granulosa cells, said culture The culture solution of follicular granulosa cells contains CNP or its variant or its analogue and HDAC (histone deacetylase) inhibitor. The present invention further provides an in vitro maturation culture solution comprising CNP or its variants or analogs thereof and HDAC inhibitors and related compositions thereof, as well as the above-mentioned medium, culture solution and composition in promoting the in vitro maturation of immature oocytes Applications.

This invention relates to a method for reducing the generation interval and advancing genetic gain in bovines via pregnancies created by oocytes harvested from select prepubertal female calves less than six months of age and / or from smaller framed bovine breeds less than ten months of age, and alternatively less than nine months of age. Said animals are treated with a hormone regiment to advance the first pubertal estrus in order to retrieve oocytes for fertilization and implantation. Oocytes are collected from said animals via laparoscopic aspiration. In the case of oocytes that have not matured in vivo, the oocytes are transferred to and cultured to maturation. Once matured, oocytes are fertilized, and the resultant embryos are transferred to synchronized recipients.