31results about How to "Increase cleavage rate" patented technology

A method is provided which allows high yields of acylated insulin. The method comprises expressing a singe-chain insulin precursor, preferably in yeast, cleaving the single-chain insulin precursor with a suitable protease which will open the peptide bond between the C-terminal amino group in the B-chain and a connecting peptide connecting the B chain with the A-chain, acylating the two-chain insulin intermediate in the ε-amino group in LysB29 and subjecting the acylated two-chain insulin intermediate to a proteolytic enzyme which will cleave of the N-terminal extension on the B- and A-chains on the precursor molecule.

The invention relates to a method for improving the developmental capacity of sheep mature oocytes after vitrification refrigeration, belonging to the technical field of biology. The method comprises the steps of micro-injection fertilization of sheep mature oocytes and activation of fertilized eggs, wherein the activation of the fertilized eggs adopts a direct current pulse-chemical activation-direct current pulse mode, and experiments prove that the embryonic developmental capacity of the vitrified sheep mature oocytes subject to is greatly improved after the vitrified sheep mature oocytes is subject to intracytoplasmic sperm injection and is activated by the activation method of the invention. Compared with the conventional external fertilization method, the cleavage rate and the blastocyst rate are respectively increased to 13.7% and 6.8%. At the same time, by comparing the chromosome ploidy of embryos subject to different treatments, the result indicates that the proportion of normal-ploidy embryos (71.5%) obtained by the ECE method is apparently higher than that (45.4%) obtained by the common chemical activation method and no obvious difference exists compared with the external fertilization groups.

The invention discloses a culture solution for improving maturity quality and development potential of in-vitro cultured porcine oocytes and its cultivating method. The culture solution is prepared from, by weight, 3-7 mu mol / L of forskolin, 3-7 mu mol / L of Roscovitine, 8-12g / L of TCM-199, 1-4g / L of NaHCO3, 0.4-0.7g / L of D-glucose, 0.05-0.15g / L of sodium pyruvate, 0.05-0.1g / L of penicillin sodium, 0.03-0.07g / L of streptomycin sulphate, 1-4g / L of polyving akohol, 0.3-0.7mu g / mL of luteotropin, 0.3-0.7mu g / mL of follicle-stimulating hormone, 8-12ng / mL of epidermal growth factor, 0.3-0.7mmol / L of L- cysteine, and 8-12% of follicular fluid. The culture solution has the advantages of improving cleavage rate, blastaea number and blastaea cell population of porcine oocytes, and effectively improving the development potential of the in-vitro cultured porcine oocytes; so the culture solution has great application prospect.

The invention relates to the technical field of pharaoh cuttlefish breeding, in particular to a breeding method beneficial to improvement of the spawning quantity and quality of pharaoh cuttlefish. The breeding method comprises the steps: (1) parent domestication, wherein adult pharaoh cuttlefish parents are put into a pond for domestication and fed with live shrimps and / or live crabs and / or livefish mixed with marine algae; (2) reproduction regulation, wherein periodic starvation and refeeding are sequentially carried out, and temperature change culture is carried out in a gradient temperature change mode; (3) spawn collection; (4) keeping of fresh and alive pharaoh cuttlefish, wherein hatching of fertilized spawns cultured on a large scale is inhibited, alive and fresh keeping is carried out by combining a C-type natriuretic peptide with cysteamine, and finally hatching and larva breeding are carried out. The method is beneficial to improvement of the physique of the pharaoh cuttlefish parents, promotes earlier and faster sexual maturation of the parents, enhances the mating probability of the pharaoh cuttlefish parents, realizes synchronous sexual maturation of the parents andimprovement of the quality of reproductive gametes, remarkably reduces the conversion ratio of normal fertilized spawns to fertilized inkjet spawns, accelerates the hatching of cuttlefish larvae through alive and fresh keeping, promotes the larval development and further improves the survival rate of larvae.

The invention provides a living cell vitrification freezing fluid and a freezing thawing method. The freezing fluid is prepared by that on the basis of an MEM culture medium and an F12 culture mediumin a volume ratio of 1:1, 50-60nmol / L of isofraxidin, 20-30nmol / L of chlorogenic acid and 40-48Mumol / L of a protective agent are added, wherein the protective agent comprises one or more of hydroxyethyl starch, tea polyphenol, 4-hydroxyethyl piperazine ethanesulfonic acid, lentinan and adenosine triphosphate. The living cell vitrification freezing fluid has the advantage that the living cell survival rate, the in-vitro fertilization and development rate, the cleavage rate and the blastocyst rate can be increased.

The invention discloses a porcine oocyte in-vitro maturation culture solution as well as a preparation method and application thereof and belongs to the technical field of oocyte in-vitro maturation culture. In order to solve the problems of low porcine oocyte in-vitro maturation rate and development rate at present, the invention provides the porcine oocyte in-vitro maturation culture solution aswell as the preparation method and application thereof. The culture solution comprises a base culture solution TCM-199, penicillin, streptomycin, NaHCO3, 4-hydroxyethylpiperazine ethane sulfonic acid, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, an epidermal growth factor, a porcine follicular fluid, pregnant mare serum gonadotropin, human chorionic gonadotropin and ramelteon. The culture solution can increase the oocyte in-vitro maturation quality, the cumulus cell diffusion index, the glutathione level, the parthenogenetic embryo blastocyst rate and blastocyst total cell number, the in-vitro fertilization embryo cleavage rate, the blastocyst rate and blastocyst total cell number and decrease the ROS level of oocytes.

The invention provides a method for improving the fertilization ability of vitrified bovine oocytes. Before the vitrification treatment, the bovine oocytes are jointly treated with a solution containing gadolinium and CLC, and then vitrified; after thawing, they are treated with methylated-β-cyclodextrin, and then fertilized in vitro. The results show that this method can significantly increase the cleavage rate and blastocyst rate of vitrified bovine oocytes, and is higher than the in vitro fertilization efficiency of fresh bovine oocytes. It has broad application prospects and is worth promoting.

The invention provides a culture method of mammal embryonic cells. During culture, uterine fetal fluid of mammals during pregnancy is not added; the culture is divided into two stages, and during the two stages, culture times and medium formulas are different. The culture method provided by the invention is suitable for culture of the embryonic cells of the multiple mammals and has higher cleavage rate, fusion rate and blastocyst rate. After an embryo obtained by the culture method provided by the invention is directly transplanted, the birth rate is high.

The invention relates to a cell nucleus operation method based on the dynamic drift modeling of the cell nucleus position, comprising the following steps: off-line calibration of the three-dimensional distribution of the cell nucleus relative to a polar body; obtaining the dynamic drift track of the cell nucleus relative to the position of a microneedle through three-dimensional finite element modeling; The factorial design method is used to determine the dominant influencing factors of the dynamic drift of the nucleus position; the relationship between the intracellular movement parameters of the microneedle and the dynamic drift trajectory parameters of the nucleus position is fitted to determine the trajectory of the microneedle required to approach the nucleus, and the microneedle is controlled by the controller. The needle moves in the cell along the determined trajectory and approaches the nucleus and completes the corresponding operations. Using the method for operating the nucleus of the present invention, the enucleation operation can automatically control the trajectory of intracellular movement and the removal amount of cytoplasm. Compared with manual enucleation, the amount of cytoplasm removed is reduced by 60% under the same enucleation success rate, and the cleavage rate of pig cloned embryos is nearly doubled.

The invention discloses a method for in vitro maturation of sheep oocytes, a pretreatment solution and a kit. The invention relates to a pretreatment solution for in vitro maturation of sheep oocytes, which is characterized in that it contains sheep small follicle fluid and conventional culture solution components for in vitro maturation of sheep oocytes. The invention also discloses a method for in vitro maturation of sheep oocytes, which is characterized in that a pretreatment step is added on the basis of the conventional in vitro maturation method. At the same time, the present invention also verifies that the cleavage rate and blastocyst rate of fertilized eggs obtained after in vitro fertilization of oocytes cultured by the maturation method of the present invention are significantly higher than those of oocytes cultured by conventional maturation methods. The pretreatment solution and the in vitro maturation culture method provided by the invention can effectively promote the in vitro maturation of sheep oocytes, and have the characteristics of no toxicity to the oocytes, less restriction, good effect and the like.

The invention discloses a pig oocyte in-vitro maturation culture solution and a preparation method and application thereof, and belongs to the technical field of oocyte in-vitro maturation culture. The problems of low in-vitro maturation rate and development rate of porcine oocytes currently are solved. The pig oocyte in-vitro maturation culture solution is prepared from a basal culture solution TCM-199, penicillin, streptomycin, NaHCO3, 4-hydroxyethylpiperazine ethane sulfonic acid, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, epidermal growth factor, porcine follicular fluid, pregnant mare serum gonadotropin, human chorionic gonadotropin and tannic acid. According to the pig oocyte in vitro maturation culture solution, the oocyte in-vitro maturation quality, the cumulus cell diffusion index, the parthenogenetic embryo blastocyst rate and blastocyst total cell number, and the in-vitro fertilization embryo cleavage rate, blastocyst rate and blastocyst total cell number can beimproved.

The invention relates to the field of animal husbandry and veterinary and particularly discloses a method for delaying the reproduction function decline of female animals, wherein the animals take in melatonin through oral administration. The delaying of the reproduction function decline of female animals is shown by increasing the litter size of older female animals, the SOD level of ovary, the SOD level of uterus, the number of useful eggs, total egg number, cleavage rate or blastocyst rate or reducing the MDA level of the ovary of older female animals, the MDA level of uterus or the number of useless eggs. In the invention, through a large quantity of objective experiments, different optimal intakes are provided aiming at reproduction function decline of specific different female animals so as to realize the best effect of delaying the reproduction function decline of female animals.

The invention provides a method for improving the moveability of sperm in frozen bull semen and the in vitro fertilization rate. The method is characterized in that when SP-TALP washing liquid is used for washing at a prior disposal stage of the frozen bull semen, or when PHE (phenylalanine) capacitation liquid is used for capacitation disposal at a capacitation disposal stage of the semen before fertilization, or when fertilization drips are prepared at an in vitro fertilization stage, glutathione and caffeine of which the purities are both higher than 98% are added; in the fertilization process of each piece of frozen bull semen, in the SP-TALP washing liquid containing bull semen or in the mixed liquid of IVF-TALP seminal fluid containing the bull semen and the PHE capacitation liquid or in the mixed liquid of the fertilization drips containing oocytes and the bull semen, the final concentration of the glutathione and the caffeine is 5mmol / L. According to the method disclosed by the invention, two substances are jointed to be respectively applied to three stages in the fertilization process of the frozen bull semen, the moveability of the sperm is greatly improved, and the fertilization rate of the sperm is improved.

The invention discloses a pig oocyte maturation culture solution in vitro. The culture solution includes insulin transferrin selenium solution ITS, fibroblast growth factor FGF, leukemia inhibitory factor LIF and inositol. When the culture medium is used for in vitro maturation of oocytes, it does not need to add serum and follicle fluid, and is safer, more stable and more efficient.