23 results about "First polar body" patented technology

The invention discloses a method for inducing ostrea rivularis triploid, and relates to the technical field of shellfish genetic breeding. The specific preparation method comprises the following steps: firstly, obtaining egg liquid and seminal fluid from gonads of female and male ostrea rivularis individuals, cleaning and curing the egg liquid, and adding the seminal fluid into the egg liquid after cleaning to obtain a fertilized egg solution; when the proportion of the first polar bodies in the eggs of the fertilized egg solution is 30-50%, adding an inducer into the fertilized egg solution for induction and cleaning to obtain induced fertilized eggs; and finally, hatching and screening the induced fertilized eggs to obtain D-shaped larvae, and culturing to obtain the crassostrea rivularis triploid population. The method for inducing the crassostrea rivularis triploid is high in triploid induction rate, high in final hatching rate, capable of stably surviving and good in application prospect.

The invention provides an oocyte in-vitro maturation culture solution additive and application thereof. The additive is tert-butylhydroquinone. By adding 100 [mu] g / mL of TBHQ into a basic culture solution, the first polar body discharge rate of oocytes is remarkably increased to 77.29%, the ROS level is reduced, and the blastocyst rate of in-vitro fertilized embryos is increased. The technology is suitable for breed conservation of livestock genetic resources such as sheep and can replace live livestock transportation to reduce the cost.

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 biology, and particularly relates to application of astragaloside in regulation of parthenogenetic activation embryo development. According to the method disclosed by the invention, the maturation rate of the oocytes can be obviously improved by adding substances such as astragaloside into the sheep oocyte in-vitro maturation liquid. The preparation method comprises the following steps: firstly, respectively adding lycopene, astragaloside, isoliquiritigenin, forsythin and baicalein into an oocyte maturation solution, and screening out the optimal dosage of each additive according to the first polar body discharge rate, the cleavage rate and the blastocyst rate; and finally, adding an additive combination into the oocyte maturation liquid according to the screened optimal dosage, detecting the first polar body discharge rate, the cleavage rate and the blastocyst rate to determine an optimal composition, and detecting the levels of ROS, GSH, mitochondrial membrane potential, oxidative stress and apoptosis related genes of the optimal composition to further determine the application effect of the composition.

The invention discloses a method for breeding land-sealed Malus salicina tetraploid. The method comprises the following steps: parent fish rearing; selecting parents; egg taking time; manually squeezing eggs; and inducing and activating, and hatching eggs. According to the method, by mastering the discharge time of the first polar bodies of the primary oocytes of the land-sealed Malus salicina and applying hydrostatic pressure induction to the first polar bodies, throwing of the first polar bodies is successfully inhibited, the first polar bodies are left in the oocytes to continue to be copied, the land-sealed Malus salicina tetraploid fry is successfully bred, and it is detected through a ploidy analyzer that 100% of the fry is tetraploid, so that the first polar bodies are completely separated from the oocytes. According to the method, the land-sealed type Malus salicina is bred to the adult fish stage at present, no malformation exists, the survival rate is high, and the method can provide theoretical and practical basis for land-sealed type Malus salicina fingerling resource protection, tetraploid population research and healthy triploid breeding, has important significance in promoting industrialization development of domestic land-sealed type Malus salicina and has wide market application prospects.

The invention discloses a method for inducing triploid of haliotis discus hannai by salinity, which comprises the following steps: (1) synchronously fertilizing sperms and ova of haliotis discus hannai to obtain fertilized ova; (2) when the appearance proportion of the first polar bodies of the fertilized eggs obtained in the step (1) is 60-70%, performing high-salt treatment or low-salt treatment on the fertilized eggs; (3) the fertilized eggs treated in the step (2) are subjected to salinity gradient sand filtration seawater re-infiltration treatment, the osmotic pressure is gradually adjusted through salinity gradient sand filtration seawater, damage to the fertilized eggs is relieved, then the fertilized eggs are resuspended in fresh sand filtration seawater with normal salinity, and conventional incubation and cultivation are conducted. According to the method, efficient triploid induction can be achieved through salinity optimization and re-infiltration steps, the problems of drug residues and equipment requirements of traditional chemical and physical methods are solved, and the method has the advantages of being safe, simple, convenient and efficient.

The invention discloses a method for improving in-vitro maturation quality and cryopreservation effect of porcine oocytes, and relates to the technical field of biology. By adding the NMN, the comprehensive efficiency of in-vitro maturation and cryopreservation of the porcine oocytes is remarkably improved. Research finds that by adding NMN into an IVM culture medium, maturation of cell nucleuses is promoted, discharge of first polar bodies of oocytes is improved, oxidative stress and lipid peroxidation of the oocytes are reduced, mitochondrial activity is improved, and cytoplasm maturation of the oocytes is facilitated. Meanwhile, after the NMN is used for pretreatment, the survival rate of the vitrified and frozen oocytes is remarkably improved, and the active oxygen level and the DNA breakage level are reduced. Therefore, the addition of the NMN in the IVM culture solution may become a new method for improving and optimizing the IVM technology, and an innovative solution is provided for pig germplasm resource preservation and excellent breeding stock breeding.

The invention discloses a novel application of a compound beta-alanine (beta-alanine) in preparation of a medicine for improving oocyte quality and treating ovarian endometriosis (OEM) related infertility, and belongs to the technical field of biological medicines. An OEM animal model proves that beta-alanine can remarkably improve reproductive function disorder of model animals, specifically, follicular dysplasia caused by endometriosis is effectively relieved, the fertility is improved, and the litter size is increased; in the aspect of oocyte quality, the compound can promote normal discharge of a first polar body in an oocyte in-vitro maturation process, reduce cytoplasm fragmentation, recover spindle body form and microtubule arrangement, enhance mitochondrial membrane potential stability and maintain the integrity of an ultrastructure. The results show that beta-alanine can reverse oocyte quality reduction caused by ovarian endometriosis in multiple dimensions, has a definite treatment potential, and provides an important candidate molecule for developing a novel assisted reproduction intervention strategy for targeting oocyte health.

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 invention discloses application of a compound shown in a formula I in promoting in-vitro cultured oocyte development or in-vitro embryo development, and in the formula I, R1 is selected from halogenated alkyl; r2 is selected from H, halogen or C1-6 alkyl; n is selected from 0, 1, 2 or 3. According to the application, the expansion area of cumulus cells in the cumulus-oocyte complex can be increased; the oxidative damage of the cumulus-oocyte complex is relieved; the first polar body discharge rate of the oocytes is improved; the abnormal distribution rate of mitochondria in oocytes is reduced; the membrane potential of mitochondria in the oocyte is improved; and the cleavage rate and blastocyst formation rate of fertilized eggs can be improved, and the apoptosis rate of oocytes can be reduced.

The invention belongs to the technical field of biology, and particularly relates to application of lycopene in sheep oocyte in-vitro culture. It is determined that the maturation rate of the oocytes can be remarkably increased by adding the lycopene and astragaloside composition into the sheep oocyte in-vitro maturation liquid. The preparation method comprises the following steps: firstly, respectively adding lycopene, astragaloside, isoliquiritigenin, forsythin and baicalein into an oocyte maturation solution, and screening out the optimal dosage of each additive according to the first polar body discharge rate, the cleavage rate and the blastocyst rate; and finally, adding an additive combination into the oocyte maturation liquid according to the screened optimal dosage, detecting the first polar body discharge rate, the cleavage rate and the blastocyst rate to determine an optimal composition, and detecting the levels of ROS, GSH, mitochondrial membrane potential, oxidative stress and apoptosis related genes of the optimal composition to further determine the application effect of the composition.

The invention discloses a novel application of myristic acid and a culture solution for promoting maturation of oocytes of pig small cavity follicles, and relates to the technical field of reproductive biology. The method can be used for preparing products for promoting maturation of the oocytes of the small cavity follicles of the pigs. The method comprises the following steps: culturing a cumulus-oocyte complex (COC) in a pig small cavity follicle under an in-vitro condition, and adding a proper concentration of MA (Maleic Anhydride) into a culture solution. By detecting functional indexes such as first polar body discharge of oocytes, mitochondrial distribution of the oocytes and apoptosis of the oocytes, it is found for the first time that in-vitro maturation of the oocytes of the small follicles is promoted and the quality of the oocytes is improved by adding MA in vitro.

The invention belongs to the field of cell culture, and particularly discloses application of dominant follicle exosomes in oocyte in-vitro maturation culture, after the dominant follicle exosomes with the concentration of 10 ng / mL-200 ng / mL are added into a porcine oocyte in-vitro maturation culture solution for porcine oocyte in-vitro maturation culture, the first polar body discharge rate is remarkably increased, and the first polar body discharge rate is remarkably increased. Therefore, the dominant follicle exosome can effectively improve the in-vitro maturity of the porcine oocytes; the dominant follicle exosomes with different concentrations are added, so that the diffusion area of cumulus cells is remarkably increased; after the dominant follicle exosome is added, the ROS fluorescence intensity is obviously reduced, and the GSH fluorescence intensity is obviously increased; by adding the dominant exosome, the remodeling of the mitochondrial structure and function in the oocyte is accelerated; adding of the dominant exosome promotes functional remodeling of the mitochondrial complex I to increase ATP generation and reduce oxidative stress so as to promote nucleoplasm co-maturation of oocytes. The subsequent embryonic development can be promoted.

The invention belongs to the technical field of shellfish genetic breeding, and particularly relates to a cultivation method of triploid offspring seeds for promoting growth of mytilus coruscus. According to the cultivation method, when the proportion of first polar bodies in fertilized eggs of mytilus coruscus accounts for 35%-40% of the total fertilized eggs, a cytochalasin B (CB) solution induces the fertilized eggs of mytilus coruscus, and then incubation and development are carried out. According to the mytilus coruscus triploid offspring seeds cultivated through the cultivation method, the triploid rate can reach 84%, the growth rate is improved by 30%-40% compared with that of diploid, larvae enter an attachment metamorphosis stage in advance, the seedling cultivation cost is greatly reduced, and a new direction is provided for industrialization development of mytilus coruscus.

The present invention relates to a method for cultivating tetraploid Haliotis discus hannai, belonging to the technical field of genetic breeding of aquaculture varieties. The method includes the selection of broodstock, the ripening promotion of broodstock, the spawning induction of broodstock, artificial insemination, and the inhibition of polar body release of fertilized eggs; the selected male and female broodstock are separately ripened; the female broodstock is induced to spawn by means of air-drying, running water stimulation, and soaking in hydrogen peroxide seawater; the eggs are preferably selected and fertilized by adding high-concentration sperm; cytochalasin B is used to inhibit the release of the first polar body of the fertilized eggs. The unique three-stage gonad ripening promotion method and spawning induction method for triploid female abalones of the present invention can obtain more high-quality eggs; the present invention uses cytochalasin B to inhibit the release of the second polar body of fertilized eggs to obtain tetraploids, with a stable induction rate of more than 96%, and a large number of tetraploid Haliotis discus hannai can be obtained.

The present invention discloses a chemical induction method for the triploid of Apostichopus japonicus Huachun No. 1, belonging to the technical field of genetic breeding of marine echinoderms. The chemical induction method comprises the following steps: promoting the maturation of broodstock, artificially inducing ovulation or sperm ejaculation, treating fertilized eggs with staurosporine, hatching the fertilized eggs to obtain juvenile sea cucumbers, and continuing to cultivate the juvenile sea cucumbers. The induction drug used in the present invention is staurosporine, which is a reversible inhibitor related to microfilament assembly and can inhibit the emission of the first polar body of fertilized eggs. Compared with traditional induction drugs such as cytochalasin B and 6-DMAP, staurosporine has the advantages of rapid inhibitory effect, low toxicity and easy cleaning. In addition, the staurosporine induction method provided by the present invention is completely different from the previous physical induction methods, not only reducing the operation difficulty, but also improving the survival rate of larvae and the triploid yield.

The invention discloses a novel application of a compound MLN4924 (Pevonedat) in preparation of a medicine for improving oocyte quality and treating related diseases, and belongs to the technical field of biological medicines. PCOS animal model experiments prove that the reproductive endocrine indexes of model animals can be remarkably improved by applying the MLN4924, and the reproductive endocrine indexes are specifically represented by restoring the regularity of an estrus cycle and reducing the levels of testosterone and luteinizing hormone in serum; meanwhile, the fertility is improved, and the birth number is increased. In the aspect of oocyte quality, the treatment can increase the number of normal oocytes in an ovary, improve the first polar body discharge rate of in-vitro maturation of the oocytes and improve the blastocyst formation rate after in-vitro fertilization. A further mechanism research shows that the MLN4924 can inhibit the plasmosis of the TRIM21 in the ovarian granular cells and down-regulate the expression level of the protein of the TRIM21. The results show that the MLN4924 provides a new drug choice for developing related diseases for treating polycystic ovarian syndrome (PCOS) and improving oocyte quality.

The application develops a quantitative induction method for tetraploid time point of Fujian oyster by taking the development stage of fertilized egg as a biological index. The application effectively eliminates the defects of poor gamete synchrony caused by traditional mixed gametes of multiple pairs of oysters by means of optimizing sperm and egg, obtaining sperm and egg by dry stripping method, and single pair fertilization. The application is different from the traditional method (after 8-10 min of fertilization, using reagent for continuous treatment for 15-20 min), and takes the development status of fertilized egg as a biological index. The time period from the beginning of fertilization of the control group to the appearance of the first first polar body (PB1) of the fertilized egg is defined as A min, and the time period from the appearance of the first PB1 of the fertilized egg of the control group to the development of PB1 of 50% of the fertilized eggs is defined as B min. When the fertilization time of the treatment group is (A-3) min, the inducer is added for continuous treatment for B min, which effectively avoids the problems of low and unstable induction rate caused by fixed induction time. The tetraploid rate of the Fujian oyster cultivated by the application is high and stable to the juvenile stage, which provides a new idea for polyploid breeding of Fujian oyster.

The present invention relates to the field of assisted reproductive technology, and particularly relates to the use of minocycline or its derivatives in promoting in vitro maturation of oocytes. Since it is important to promote nuclear and cytoplasmic maturation during the process of oocytes resuming meiosis in vitro in the IVM technology, cytoplasmic maturation is the key factor affecting embryonic developmental potential. The research of the present invention has found that the application of MiH in the IVM culture medium not only facilitates nuclear maturation and helps improve the extrusion of the first polar body of oocytes, but also balances the damage of ROS to cytoplasmic organelles and the nucleus, which is beneficial to the maturation of oocyte cytoplasm. At the same time, it improves the blastocyst formation rate of fertilized eggs of aged mouse oocytes. Adding MiH to the IVM culture medium may become a new method to improve and optimize the IVM technology, which is of great significance for improving the fertility of elderly women and the development of clinical reproductive medicine.

The invention discloses an artificial propagation-hybridization combined breeding method for meat goats, and relates to the technical field of animal breeding, and the method comprises the following specific steps: selecting a goat variety with good meat performance as a female parent, and selecting a goat variety with large physique as a male parent; the method comprises the following steps: performing in-vitro culture on female parent oocytes to obtain mature oocytes containing first polar bodies, screening high-activity sperms by adopting a temperature gradient screening device, performing in-vitro fertilization on the mature oocytes and the high-activity sperms, culturing the mature oocytes and the high-activity sperms to a hybrid blastocyst form, transplanting the mature oocytes and the high-activity sperms into a female body, and developing the mature oocytes and the high-activity sperms until the mature oocytes and the high-activity sperms are naturally delivered to obtain a hybrid F1 generation; backcrossing the F1 generation with the female parent, backcrossing the backcrossed offspring with the female parent for multiple times until a breeding target is met, and carrying out cross-crossing fixation to obtain a new variety / line with excellent characters; the sperm screening efficiency is improved by utilizing the screening device, and the quality of the screened sperms is ensured, so that the fertilization efficiency and the embryo survival rate are remarkably improved.

The application discloses a method for inducing triploid of Haliotis discus hannai Ino by salinity, comprising the following steps: (1) synchronously fertilizing sperm and ovum of Haliotis discus hannai Ino to obtain fertilized eggs; (2) when the proportion of the first polar body appearing in the fertilized eggs obtained in step (1) is 60-70%, the fertilized eggs are subjected to high-salt treatment or low-salt treatment; and (3) the fertilized eggs treated in step (2) are subjected to reperfusion treatment of salinity gradient sand-filtered seawater, so as to gradually adjust the osmotic pressure by the salinity gradient sand-filtered seawater, slow down the damage to the fertilized eggs, and then resuspend in fresh sand-filtered seawater with normal salinity, and perform conventional incubation and cultivation. The application can realize efficient induction of triploid by optimizing the salinity and reperfusion steps, avoids the problems of drug residues and equipment requirements in traditional chemical and physical methods, and has the advantages of safety, simplicity and high efficiency.

The invention discloses a novel application of palmitoleic acid and a culture solution for promoting in-vitro development and maturation of porcine oocytes, and belongs to the field of reproductive biology in developmental biology. The method can be used for preparing products for promoting in-vitro development and maturation of oocytes. According to the present invention, pig cumulus-oocyte complexes (COCs) are cultured under the in vitro condition, and the palmitoleic acid is added to the culture solution to explore the influence of the palmitoleic acid on the first polar body discharge rate, the cortical particle distribution, the mitochondrial distribution and the oocyte reactive oxygen species (ROS) level of the oocytes, such that the results show that the in vitro addition of the palmitoleic acid can improve the oocyte quality, and the in vitro addition of the palmitoleic acid can provide the research on the oocyte first polar body discharge rate, the cortical particle distribution, the mitochondrial distribution and the oocyte reactive oxygen species (ROS) level; and a good effect is achieved.

The invention relates to the technical field of assisted reproduction, in particular to application of minocycline or a derivative thereof in promoting in-vitro maturation of oocytes. Due to the fact that in the IVM technology, it is very important to promote cell nucleus and cytoplasm maturation in the in-vitro meiosis recovery process of oocytes, cytoplasm maturation is the key to influence the embryonic development potential, and it is found through research that when MiH is applied to an IVM culture medium, cell nucleus maturation is facilitated, and discharging of first polar bodies of the oocytes is improved; moreover, the damage of ROS to organelles and cell nucleuses in cytoplasm is balanced, and maturation of ovum cytoplasm is facilitated. Meanwhile, the blastocyst formation rate of fertilized eggs of oocytes of aged mice is improved. The method for adding MiH into the IVM culture solution can become a new method for improving and optimizing the IVM technology, and is of great significance to improvement of fertility of elderly women and development of clinical reproductive medicine.