167 results about "External fertilization" patented technology

The invention relates to a method for establishing a ferret model capable of applying to study human disease and an application thereof, which belong to the biology field and specifically relates to a ferret ovulation induction technology and a ferret external fertilization technology. With the method for establishing the ferret model based on the technology of utilizing CRISPR/Cas9 on the above aspects, the method can be applied to study human diseases.

The invention discloses a freezing method of pig sperms. Unexpectedly, the DNA (deoxyribonucleic acid) completeness and the in vitro fertilization (IVF) rate of unfrozen pig sperms can be obviously improved by using 9(w/v)% of low-density albumin for treatment during the freezing process.

Ovarian germ-line-competent embryonic stem cells (GLC-ESC) are cultured, either in the presence or absence of a compound having estrogenic activity. The GLC-ESC are either collected prior to specific commitment or are permitted to remain in the culture medium for a time sufficient to develop into oocytes, and the oocytes may be fertilized by adding sperm to the culture medium. The fertilized oocytes may be permitted to develop into embryos, which may be transferred into the uterus of an adult human female or frozen for later use. The invention provides a method for obtaining by in vitro fertilization an embryo that is genetically related to a human female who is not producing oocytes.

The invention discloses a high density freezing maintain pig seminal fluid method with fistula and product in animal propagation technical domain, which comprises the following steps: collecting fresh seminal fluid; detecting; pre-treating the seminal fluid; defreezing; proceeding assessment procedure of defreeze seminal fluid; diluting with freezing diluting liquid; storing with high density super low temperature with 0. 25mL fistula; stabilizing the defreezing sperm above 0. 5 and average active at 0. 58; reaching the highest at 0. 68; proceeding pig external fertilization with the sperm; getting 54. 3% cleavage ratio and 48. 5% morula developing ratio; meeting one sperm dosage with sperm quantity of one 0. 25mL fistula. This invention possesses larger using latent force and market developing prospect.

The invention provides a sperm screening and in vitro fertilization bionic device based on micro fluidic technology. The sperm screening and in vitro fertilization bionic device based on micro fluidic technology comprises a bionic device main body; the bionic device main body is composed of a glass sheet layer, a silica gel pipeline layer, and a polydimethylsiloxane layer; a center cell is arranged at the middle part of the silica gel pipeline layer; an inlet cell, a first outlet cell, and a second outlet cell are arranged outside the center cell; the center cell is communicated with the inlet cell, the first outlet cell, and the second outlet cell via a first pipeline, a second pipeline, and a third pipeline respectively; the first pipeline is provided with a first sample adding port; the second pipeline is provided with a second sample adding port; and the third pipeline is provided with a third sample adding port. The sperm screening and in vitro fertilization bionic device is used for stimulating complete natural impregnation process, unfavorable influences of operation on gametes and embryos are avoided, and polyspermy is avoided; high quality embryos at stimulated natural impregnation states can be obtained, in vitro fertilization success rate is increased greatly, the potential causes of sterility can be identified, and a direction is provided for clinical treatment.

The invention provides an external fertilization and embryo transplantation information checking method. The method comprises the following steps that (1) identity information corresponding to a sperm, an ovum, an embryo and a matrix is stored in a database; (2) relevant identity information of culture dishes which carry the sperm, the ovum and the embryo and relevant identity information of a matrix identity recognition ring are recognized through the RF technology; (3) searching is performed on the database to compare whether the obtained identity information is matched with the database or not, and pairing is correct if yes; if not, pairing is wrong. According to the external fertilization and embryo transplantation information checking method, the defects in the prior art are overcome; under the condition of no manual intervention, the identity information of the culture dishes which carry the sperm, the ovum and the embryo and the identity information of the matrix identity recognition ring are recognized through the RF technology during external fertilization and embryo transplantation, and the recognized identity information is compared. In this way, the accuracy of the external fertilization and embryo transplantation can be guaranteed.

The invention relates to a culture solution for bovine IVF and a method for improving bovine IVF. The method for bovine IVF comprises the following steps: washing a mature cumulus-oocyte complex in afertilization culture solution, transferring the mature cumulus-oocyte complex into the fertilization culture solution, and putting the fertilization culture solution into an incubator for later use;taking a frozen seminal tubule from liquid nitrogen, performing unfreezing in a water bath at 37 DEG C, injecting seminal fluid into a centrifuge tube containing a seminal fluid preparation culture solution, and discarding a supernatant after centrifugation; adding the seminal fluid preparation culture solution into the centrifuge tube, resuspending sperm precipitate, and taking a proper sperm suspension for sperm counting; and adding the sperm suspension with the calculated volume into fertilization culture liquid drops containing oocytes, placing a culture tray into the incubator, and carrying out sperm-ovum co-incubation to complete IVF operation for embryo in-vitro culture. The fertilization culture solution and the seminal fluid preparation culture solution contain sodium pyruvate, heparin and other components. The method can significantly improve the efficiency of bovine IVF.

The invention relates to a method for bovine in-vitro fertilized embryo culture and a used culture solution. Particularly, on one hand, the invention relates to a method for culturing a bovine in-vitro fertilized embryo. The method comprises the following steps of putting the bovine in-vitro fertilized embryo into a bovine in-vitro fertilized embryo culture solution and carrying out embryo in-vitro culture under the conditions that the temperature is 38.5 DEG C, the content of CO2 is 0.5% and the humidity is 100%. The invention further relates to a bovine in-vitro fertilized embryo culture solution. The culture solution comprises NaCl, KCl, NaHCO3, MgCl2, KH2PO3, sodium pyruvate, glucose, calcium galactonate, fetal calf serum, L-glutamine, an essential amino-acid, a nonessential amino acid, glutathione and water. The method and the used culture solution have excellent technical effects in the specification, for example, the method has a higher blastocyst hatching rate when the bovine in-vitro fertilized embryo is cultured.

Sperm mobility and impregnation of an oocyte is enhanced by placing sperm in an aqueous solution of phospholipids prior to in vitro fertilization or artificial insemination. The aqueous phospholipid solution can also be used during storage or cryopreservation of the sperm. Also, the motility of sperm produced or ejaculated and the environment into which it is placed is enhanced my ingestion of the phospholipid composition by the male or female, or a vaginal placement of compositions containing the phospholipids.

The invention discloses an in vitro maturation culture method of denuded oocytes of mice. The method comprises the following steps: (1) performing in vitro maturation culture on the denuded oocytes; (2) performing parthenogenetic activation on matured denuded oocytes; (3) performing in vitro fertilization on the matured denuded oocytes. According to the method, an environment for artificially simulating natural maturation of oocytes can be created, so that the denuded oocytes can efficiently maturate and grow in vitro and can be subjected to in vitro fertilization and embryonic development as same as normally growing oocytes after in vitro maturation culture. The practice proves that the in vitro maturation culture efficiency of the denuded oocytes of the system is high, the system is mature and stable, and the embryonic development quality of the denuded oocytes is high after in vitro natural fertilization, so that the application and popularization of the denuded oocytes in reproductive health of human beings and genetic breeding of modern livestock can be greatly facilitated. In addition, the operation of obtaining blastulae from the denuded oocytes is simplified and the denuded oocytes serving as ovum sources can be conveniently put into actual production.

The invention provides a method for improving the blastocyst rate of in-vitro embryos of animals. According to the method, the developmental capacity of a parthenogenetic embryo and an in-vitro fertilization embryo can be remarkably improved by adding type I interferon into an embryonic development culture solution for the in-vitro culture of the embryos; compared with the blastocyst rate obtained under the culture of the conventional culture solution, the blastocyst rate is improved by 7 to 13 percent; and the blastocyst hatching rate is also remarkably higher than the blastocyst hatching rate under the culture of the conventional culture solution. By the method, the early development capacity of in-vitro embryos of cattle and sheep is improved, so that the in-vitro production efficiency of embryos of the cattle and sheep is improved, and high-quality embryos are provided for the scientific research and production field in which the embryos of the cattle and sheep are taken as materials.

The invention provides a two-layer percoll density gradient centrifugal separation method of boar sperms. The method is suitable for normal sperm specimen and sperm specimen with low sperm density or low sperm motility, and is a simple, convenient and fast sperm treatment method. The method can be used for remove leucocytes, bacteria, fragments and anti-sperm antibodies in the sperms and improving the delayed sperm liquefaction, the sperm in-vitro capacitation and the like, so that the sperms can effectively fertilize ova to improve the pregnancy rate, and the percentage of the sperms with motility and normal shape in the obtained sperms is about 95%. The method provided by the invention is suitable for screening sperms with motility from fresh boar sperms and diluted sperms, thus providing high-quality sperms for the study of boar sperm capacitation mechanism and in-vitro fertilization.

The invention discloses a swine in-vitro embryo culture solution. The swine in-vitro embryo culture solution disclosed by the invention is obtained through adding ligustrazine, with the final concentration of 0.05-5.00 microgram/milliliter, into an embryo culture solution NCSU-23. A traditional Chinese medicine monomer component, namely ligustrazine, is added into the swine in-vitro embryo culture solution disclosed by the invention; shown by experiments, through carrying out in-vitro culture on the swine parthenogenetic activated embryos and the swine in-vitro fertilized embryos by using the culture solution, the development efficiency of the embryos and the number of the cell masses and total cells in blastulae can be effectively increased. The swine in-vitro embryo culture solution has the advantages that a foundation is laid for the development of other biotechnologies, medical research and animal husbandry, and meanwhile, a certain theoretical basis for clarifying the development mechanism of early swine embryos is provided.

The invention discloses a culturing device, a culturing liquid and a culturing system for the external fertilization of human ovums and sperms and the early development of an embryo. The culturing device comprises at least two thermostatic culturing chambers, two small-sized air bottle chambers, a mixed gas filtering device, an air pressure protecting device, a mixed gas thermostatic preheating chamber, an air supplying pipe and the like, wherein each culturing chamber is a confined space with the volume of at least 100ml. The culturing liquid contains sodium chloride, magnesium sulfate, monopotassium phosphate, sodium bicarbonate, ethylenediamine tetraacetic acid, taurine, sodium pyruvate, L-lactic sodium lactate, glucose, gentamicin, alanine-glutamine dipeptide, 18 types of L-lactic amino acids except alanine and glutamine, and the like. When the culturing device and the culturing liquid are used, the external fertilization of the human ovums and sperms and the whole process of the development of the embryo before embryo implantation can be finished; the high-quality blastocyst forming rate and the embryo implantation rate are obviously increased.

The invention relates to a progestin nano composition, an applying pellet containing the progestin nano composition, a method for preparing the progestin nano composition and the applying pellet, preparations containing the progestin nano composition and/or the applying pellet such as capsules, tablets and lyophilized powder, and application of the progestin nano composition and the applying pellet on preparation of medicines. The medicines are used for treating and/or preventing diseases caused by hormone deficiency, or are used for hormone replacement therapy after external fertilization.

The method of producing extracorporeal buffalo embryo in controlled sex includes recovering buffalo egg in butchery or from living buffalo body, extracorporeal culturing to mature, external fertilization with fresh or frozen X or Y sperm of fine breed bull in a flow cytometer; and recovering embryo of the fertilized egg via external culture to blastula stage for transplanting or further freeze maintaining. The embryo is used in generating posterity of the expected sex. The present invention has the advantages of industrial embryo production in controlled sex and raised propagation speed of high yield milk buffalo.

The invention provides a method for fertilizing oocytes in vitro of young stock twice, which comprises the following steps of: performing FSH super-ovulation processing on 45 to 100-day young female animals which are receptors and taking more than 100 oocytes from each young female animal for later use; mixing the oocytes in vitro matured for 25 to 26 hours and sperms to finish in-vitro fertilization of the first time; and after 5 to 6 hours, performing the in-vitro fertilization of the second time, transferring the fertilized oocytes into a culture plate for culture after 13 to 14 hours, and counting a clearage rate after 48 hours. The matured oocytes need treating with 0.1 percent hyaluronidase for 30 to 60 seconds, and then are added into a fertilization medium for incubation after cumulus cells are partially removed from the oocytes in a blowing and sucking way, wherein the concentration of the fertilization medium is 50 to 70 mu l per drop and is well balanced for 2 to 5 hours; semen is unfrozen with a water bath and is placed into a CO2 box, and after 20 to 25 minutes, supernatant fluid is extracted and centrifugated for 2 to 5 hours, and then the precipitated sperms are added into the fertilization medium for incubation according to the density of 2-4*106 sperms per ml; ova fertilized for 5 to 6 hours are taken out and are placed into the fertilization medium; the sperms are unfrozen by the similar method, are added into the fertilization medium, and after 13 to 14 hours, are washed and transferred into a well-balanced four-hole culture plate for culture, wherein each hole of the four-hole culture plate contains 500 mu l of the washed solution. The method has the advantages of simplicity, practicability and the capacity of making the fertility rate of young cows and ewes reach 80 to 92 percent while blastocyst rate reach 40 to 50 percent.

A safe recording and comparison method for artificial fertilization, in-vitro fertilization and embryo transfer comprises the following steps of: step 1, archiving of a male parent and a female parent: storing identity information, pairing information and biological feature information; step 2, semen collection: identifying male parent information at first, then collecting semen, and establishing a matching relation between the semen and the male parent; step 3, semen treatment: during treatment, checking whether the semen is matched with the male parent in each sample transfer or transmitting the matching relation; step 4, ovum collection: identifying female parent information at first, then collecting an ovum, and establishing a matching relation between the ovum and the female parent; step 5, ovum fertilization: identifying semen and ovum information at first, then checking whether pairing is allowed, and if so, continuing, or otherwise, sending out an error prompt; and step 6, transplantation: verifying the female parent information at first, then checking whether an embryo is matched with the female parent, and if so, continuing, or otherwise, sending out the error prompt. The checking is performed before every operation to prevent errors.