66 results about "Maturation oocyte" patented technology

The invention belongs to the technical field of pig genetic breeding, and specifically relates to a culture solution capable of improving in vitro maturation rate of pig oocytes and application thereof. The culture solution capable of improving the in vitro maturation rate of pig oocytes is prepared from the following raw materials with the following concentrations: 9.5 g/L of TCM-199, 2.2 mg/mL of NaHCO3, 3.05 mmol/L of D-glucose, 0.91 mmol/L of Sodium Pyruvate, 0.5 g/L of Streptomycin sulfate, 0.05% of PVA, 0.57 mmol/L of Cysteine and 0.075 g/L of Penicillin G. The 0.05% of PVA and 5% of a follicular fluid are added on the basis of pig oocyte in vitro maturation culture solution widely used in our country, so that the in vitro maturation rate of the pig oocytes can be improved to more than 90%, and matured oocytes with higher quality and more quantity can be provided for porcine somatic cell nuclear transfer technology.

The invention discloses a method for promoting in-vitro maturing of human immature oocyte by utilizing a 3D printing technology. The method comprises the following steps: separating and collecting granular cells in clinic, propagating the granular cells in an in-vitro culture mode for 2 to 7 days; evenly mixing biological hydrogel with the pre-processed granular cells, printing artificial follicles by a biological 3D printing technology; adding a culture liquid into the artificial follicles, wherein the volume percentage content of human mature follicular fluid of the culture liquid is not less than 10%; culturing for 24 to 48 hours, then adding an immature oocyte-cumulus granular cell composite body, and culturing the 3D cells for 24 to 48 hours to promote the maturing. In the provided method, a 3D cell printing technology is adopted, human granular cells are used to construct an immature oocyte 3D culturing system, and multiple growth factors and simulated follicle culture liquid are added at the same time to create a simulated human follicle so as to promote the in-vitro maturing of immature oocyte.

The invention provides a purpose for preparing supplementary reproduction medicine from the following raw material medicine in percentage by weight: 18 to 30 parts of radix rehmanniae preparata, 9 to 12 parts of dogwood, 8 to 16 parts of Chinese yam, 9 to 12 parts of fructus lycii, 8 to 16 parts of semen cuscutae and 8 to 16 parts of antler gum. The invention also provides a method for promoting ovarian ovum in vitro maturation. The method comprises the following steps that a, an immature ovarian ovum is taken; and b, the immature ovarian ovum is placed in M199 culture liquid, a microdrop method is adopted for culture for 24 hours, and a mature ovarian ovum is obtained, wherein 2 percent of medicine is contained in the culture liquid. The medicine can promote the growth and development of the immature ovarian ovum in vitro and mainly promotes the ovarian ovum core maturation, and in addition, the cytoskeleton abnormality probability of the mature ovarian ovum is not increased. The medicine can also generate the influence on the ovum passing capability of sperms, and the reliable basis is provided for clinically preventing and treating the male sperm dysfunction, improving the IVF (in vitro fertilization) success ratio and reducing the use rate of ICSI (intracytoplasmic sperm injection).

The invention relates to cell culture and particularly discloses an immature oocyte culture scheme which can promoting in-vitro maturation of immature oocytes. Specially, the scheme adopts follicular granulosa cells and a culture solution for culturing the follicular granulosa cells, wherein the culture solution for culturing the follicular granulosa cells contains CNP or its variants or analogues and an HDAC (histone deacetylase) inhibitor. The invention further provides an in-vitro maturation culture solution containing the CNP or its variants or analogues and the HDAC inhibitor and related compositions and application of a culture medium, the culture solution and the compositions in promotion of in-vitro maturation of immature oocytes.

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 invention discloses a simple in-vitro maturation culture method for oocytes, which comprises the following steps: A1, collecting oocytes; A2, preparing a closed air bag and performing in-vitro maturation culture of oocytes; and A3, performing parthenogenetic activation and in-vivo culture of oocytes, namely after 22 to 24 hours of in-vitro maturation culture, selecting mature oocytes discharging first polar bodies, activating by a chemical process in the 28th hour, culturing for 3 days in merogenesis culture solution, culturing for 4 days in blastaea culture solution, and observing and counting a blastaea rate after 7 to 9 days. When a self-prepared closed air bag system is used, the in-vitro culture of the oocytes and embryos can be accomplished by using a common biochemical culture tank or water bath pot, so the dependency on expensive instruments is reduced and a research doorsill is reduced. When the simple closed air bag is used for in-vitro culture of the oocytes, blastaea can be grown, and the in-vitro maturation rate, merogenesis rate and blastaea rate are higher than those of oocytes cultured in a normal CO2 culture tank.

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.

Methods and kits are provided for assessing the ovarian reserve and predicting the ovarian response to fertility treatments in a female subject. The serum levels of MIS are shown to be positively correlated with the production and retrieval of mature oocytes and serve as prognostic indicators for the female response to fertility treatment. The MIS levels can be monitored prior to and during fertility treatment and are useful to adjust the timing and dosage of treatments in order to produce optimal outcome in individual patients, to avoid ovarian hyperstimulation, or to indicate cancellation of an unsuccessful treatment. MIS can also be administered to women to stimulate follicle development and to prevent depletion of ovarian reserve.

The invention discloses a method for improving transferring efficiencies of pig somatic cell nucleuses. The method includes the following steps of selecting and cultivating egg mother cells; separating and cultivating donor cells; using a blind aspiration method to remove a mature egg mother cell nucleus and a first polar body, injecting a donor cell to perivitelline space, subjecting a reconstructed embryo to electrofusion, and activating the reconstructed embryo; and placing the reconstructed embryo in a PZM-3 culture solution containing RG108 with concentration of 100-200 mu M and Scriptaid with concentration of 100-500 nM to be cultivated for 14-16 hours, and then placing the reconstructed embryo in a PZM-3 culture solution for cultivation. According to the method, development blastosphere rates and blastosphere qualities of pig cloning early embryos can be obviously improved; and simultaneously, two epigenetic modification of gene methylation and acetylation with close connection and synergistic effects can be adjusted, the method is better in effects than that only by using the Scriptaid for treatment, early genetic expression of embryos can be adjusted to be normal, and finally the transferring efficiencies of the pig somatic cell nucleuses are improved.

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 belongs to the field of assisted reproduction and particularly relates to vitrification refrigerating liquid. According to the vitrification refrigerating liquid provided by the invention, taxol is added to serve as a cryoprotectant and the vitrification refrigerating liquid obtained with reference to the taxol adding mode is treated, so that the development rate of in vitro fertilized blastocyst after the oocyte is thawed can be increased. The taxol serving as an additive in the refrigerating liquid can achieve the effects of reducing freezing injury of the mature oocytes and improving the development capability of the embryo after fertilization. The optimal concentration of the adding mode is obtained by researching the adding concentration of the taxol. The vitrification refrigerating liquid provided by the invention can greatly increase recovery rate after the frozen mature oocytes are thawed and the development rate of the blastula and achieves the protective effecton the cell ultrastructure.

The invention discloses a culture solution for promoting in-vitro maturation of porcine oocyte, the culture solution takes an M199 culture solution without HEPES as a basic culture solution, and porcine follicular fluid, L-cysteine, sodium pyruvate, epidermal growth factor, insulin, gonadotropin, chorionic gonadotropin and WNT5A cytokine are also added. When the culture solution is used for carrying out in-vitro culture on the porcine oocytes, the in-vitro maturation efficiency and quality of the oocytes can be effectively improved, the obtained mature oocytes are used for carrying out porcinein-vitro fertilization and somatic cell nuclear transfer embryo production, and the blastocyst development rate and quality of the mature oocytes are also remarkably improved.

The invention discloses a method for improving the parthenogenetic development potential of vitrified mature oocytes, wherein oocytes are frozen in a melatonin-containing freezing liquid, and then arethawed with a melatonin-containing sucrose solution, the thawed oocytes are cultured, the cultured oocytes are subjected to parthenogenetic activation, and finally the parthenogenetically activated embryo is cultured in vitro. According to the present invention, the method has advantages of simple process and short time consumption, and can effectively improve the parthenogenetic activation development potential of vitrified mature oocytes.

The invention provides an extraction method of a mouse ovarian tissue fluid. The method comprises the following steps: placing mouse ovaries in a basal medium; mashing and dissloving the ovarian tissue fluid in the basal medium to form a pre-culture solution. The invention also provides an in-vitro maturation culture method of mouse oocyte, and the method comprises: transferring an immature mouse oocyte into a mature culture solution for in-vitro culture, wherein the mature culture solution is formed by adding the pre-culture solution in the basal medium of oocytes. In addition, the invention also provides a method for obtaining a test-tube mouse through embryo transplantation, and the method comprises: carrying out in-vitro fertilization and embryo transplantation on the oocyte matured by the in-vitro maturation culture method. By using the ovarian tissue fluid provided by the invention in the in-vitro maturation of the mouse oocyte, the cleavage rate of the in-vitro fertilized mature egg can be effectively increased, and is nearly the same as the that of the in-vivo mature egg; and finally, the test-tube mouse can be obtained through embryo transplantation.

The invention discloses a method for evaluating the influences of smoking on the quality of oocytes. Specifically, oocytes are cultured to be mature in culture solutions containing different concentrations of cotinine. Finally, the influences of smoking on the quality of oocytes are evaluated by detecting some indicators, such as the maturation rate of oocytes, chromosomal aneuploidy, actin distribution, mitochondrial membrane potential, intracellular reactive oxygen species level, spindle morphology, chromosome arrangement of mature oocytes, sperm binding number after in-vitro fertilization,pronucleus formation rate after parthenogenetic activation and the like. Through a large number of objective tests, the influences of smoking on the quality of oocytes are evaluated, thus providing areference of theoretical data for the maintenance of reproductive capacity of female smokers.

The invention belongs to the field of animal breeding and particularly relates to an artificial reproduction-hybridization joint breeding method for a meat goat. The method comprises the steps of selecting one goat variety with good meat performance as a female parent, selecting another goat variety with a large body size as a male parent; culturing oocytes of the female parent in vitro into mature oocytes containing a first polar body, and soaking semen of the male parent with a semen motility enhancement liquid and then preparing a semen suspension; carrying out in vitro fertilization on themature oocytes and the semen suspension, culturing zygotes into a hybrid blastocyst form, transplanting hybrid blastocysts into a body of the female parent for developing to natural delivery in the body of the female parent and obtaining hybrid F1; and backcrossing the hybrid F1 and the female parent, backcrossing backcross descendants and the female parent for multiple times until a breeding objective is met, and carrying out crossbred fixing to obtain a new variety/line with good characters. According to the method, an artificial assistant reproduction technology is combined with a traditional hybridization technology, so that the number of animal embryos can be increased, the survival rate of the animal embryos can be improved and the method has a good application prospect in the fieldof animal breeding.

The invention belongs to the field of biology, and particularly relates to a method for lamb oocyte in-vitro fertilization culture. The method comprises the steps of oocyte acquisition and maturation culture, sperm capacitation, mature oocyte in-vitro fertilization and zygote culture. During sperm capacitation and mature oocyte in-vitro fertilization, incubating time, the rotating speed of a centrifugal machine, centrifugation time, the number of times of centrifugal washing and mature oocyte culture time conditions are optimized, so that the zygote cleavage rate of lambs is basically the same as that of adult ewes.

The invention relates to the technical field of embryo engineering, in particular to a method for obtaining male pronucleus by using xenogeneic animal oocytes as a medium. The method comprises the following steps of step 1, obtaining mature oocytes of animals A, with a first polar body, and placing the obtained mature oocytes in external fertilization liquid droplets of animals B; step 2, injecting the sperms of the animals B into the liquid droplets containing the oocytes in the step 1 to obtain sperm and oveum mixed droplets; and step 3, placing the sperm and oveum mixed droplets obtained inthe step (2) in a CO2 incubator, and performing hatching until sperms of the animals B enter the cytoplasm of the oocytes to form male pronucleus, wherein the animals A and the animals B are xenogeneic animals. According to the method, a fertilization biology principle of reproductive isolation of uncrossed fertilization of the xenogeneic animals is used, an external fertilization method is adopted, the oocytes of the xenogeneic animals are used as a medium to obtain the male pronucleus of another animal, and the formation rate of the male pronucleus is as high as 17.8% or above.

The invention discloses novel use of a Wuzi Yanzong prescription. The invention provides use of a composition in preparation of complementary medicines before or/and after transplantation of ovaries. The composition comprises the following active pharmaceutical ingredients in parts by weight: 30-50 parts of fructus lycii, 30-50 parts of semen Cuscutae, 15-25 parts of raspberry, 3-7 parts of schisandra chinensis and 7-13 parts of semen plantaginis. The study shows that a Wuzi Yanzong pill is used before or/and after transplantation of ovaries, is capable of promoting growth and development of autologously transplanted ovarian follicles after being frozen and stored, restoring the developmental capacity of autologously transplanted follicular oocytes, promoting maturing of immature oocytes which are cultivated in vitro, improving the 2-cell embryos development rate and mulberry embryo development rate for in vitro development, has a good protective effect on functions of the frozen autologously transplanted ovaries, and is capable of effectively increasing the success rate of ovarian transplantation, and providing new supplementary means for an ovarian transplantation operation.