66 results about "Ovarian Follicle Fluid" patented technology

A device and method for assessing characteristics of follicular fluid in vivo comprises an aspiration needle for oocyte retrieval having one or more sensors disposed on its outer cannula wall or embedded in its outer cannula wall for sensing and measuring characteristics of follicular fluid constituents such as pH level and oxygen concentration. The sensors are microsensors that communicate through a physical connection or wirelessly with a meter and display. The measurements are recorded and optionally printed for later review and selection of the most viable oocytes. The method involves inserting into an ovary an aspiration needle with one or more sensors. The needle is guided to a first follicle and inserted in the first follicle. Characteristics of the follicular fluid are sensed and measured and accordingly displayed on the meter and recorded for later reference and selection. The fluid is collected into a first collection container. The sensors can then be reset to a reference baseline and then additional follicles can be aspirated and measured in the same manner. After collection, the measurements of follicular fluid characteristics for each follicle can be assessed and the most viable oocytes can be selected.

The invention discloses a porcine oocyte in vitro maturation medium and methods of preparation and culture. The medium comprises a TCM-199 base medium and the added porcine follicular fluid, fetal bovine serum, eCG, hCG, EGF and IGF-1. The porcine oocyte in vitro maturation medium of the invention can significantly increase the rate of maturation of porcine oocytes in vitro. Experiments confirm that after the oocyte collection and 42-44h maturation, the medium of the invention can have a polar body rate stabilized at 80% and a parthenogenetic embryo development rate up to 85%, and the entire maturation process does not require replacement of the medium and is easy to operate.

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.

Concentrations of endogenous steroids in ovarian follicular fluid are used to develop steroid profiles which provide means for the diagnosis and prognosis of endocrine-related conditions and for identifying and developing appropriate treatments for related conditions, including the identification and development of suitable protocols for in vitro fertilization (IVF), treatment and predictive strategies for successful IVF outcomes and selected uses of oocytes for IVF or embryonic stem cell procedures.

The invention discloses an effect study method of melatonin in prediction of ovarian reserve and IVF-ET (in-vitro fertilization and embryo transfer) outcome. The method includes: collecting basic clinical data of 61 females, and dividing the 61 females into three groups, namely a poor ovarian response group, a normal ovarian response group and a high ovarian response group mainly through age, retrieved oocyte number and anti-mullerian hormone value according to the stimulus response of the ovaries; collecting follicular fluid of the 61 females, adopting melatonin radioimmunoassay to determine melatonin concentration in the follicular fluid of the 61 females, and analyzing the levels of the melatonin concentration and other clinical data in the poor ovarian response group, the normal ovarian response group and the high ovarian response group; analyzing the levels of relevant data of in-vitro fertilization outcome in the poor ovarian response group, the normal ovarian response group and the high ovarian response group, and verifying relevance of the melatonin concentration and IVF-ET outcome; verifying relevance of the melatonin level and ovarian reserve. Through detection of melatonin concentration in the follicular fluid, ovarian reserve and IVF-ET outcome are predicted.

The present invention relates to the competence of oocytes to fertilization, uterine implantation and development into a living being. The invention describes ovarian markers whose expression is predicative of oocyte competency that are detected and / or measured in follicular fluid, cumulus cells and / or follicular cells of a mammal. Also described are methods for evaluating competence of mammalian oocytes, methods for selecting a mammalian oocyte for assisted reproduction (AR), and screening methods for identifying stimulatory or inhibitory compounds to mammalian oocyte competence.

The present invention belongs to the field of auxiliary reproduction technology. Said invention described culture solution containing human mature follicular fluid is used for external culture technique of human immature ovum, and its composition is formed from Ham'e F-10 or TCM199, human serum substitute SSS, rFSH, HCG, 17 beta-estradiol and human mature follicular fluid. The in-vitro mature (IVM) technique of immature ovum can be used to take out the immature ovum of sterility woman with ovum maturation disturbance, and the culture solution prepared by said invention is used to culture it, and make it mature and make in-vitro fertilization and embryo transplantation to help gestation. It can obtain 65% of ovum mature rate and 33.3% of gestatino rate.

The invention provides a method for enhancing the ectogenesis of sheep oocyte. The method adopts an adenylate cyclase activator Forskolin (FSK), phosphodiesterase inhibitor cilostamide (CIL) and 3-isobutyl-1-methylxanthine (IBMX), treats the sheep oocyte in a combining manner and improves the capability of the ectogenesis of the sheep oocyte. The method comprises the following detailed steps of: picking the ovary of the killed sheep, putting the ovary of the killed sheep into normal saline for cleaning for 3-4 times; pumping the 2-8mm follicles on the surface of the ovary by using a 10ml syringe which absorbs 1ml ovum-pumping liquid containing 100mu mol / L FSK and 500mumol / L IBMX and is provided with a number-9 needle head, and injecting the follicular fluid recovered in the syringe into a35-60mm vessel; picking the oocyte under a microscope, putting the oocyte into the pumped follicular fluid, cleaning the oocyte for three times with in-vitro mature fluid containing 20mu mol / L CIL, and putting the oocyte in a CO2 incubator for culture; then cleaning the oocyte for three times with in-vitro mature fluid which does not contain CIL, putting the oocyte in the CO2 incubator for culture and adding into in-vitro fertilization fluid drop; unfreezing the seminal fluid with water bathing, and commonly incubating with the oocyte; and calculating the cleavage rate after 24 hours and calculating the blastocyst rate after 168 hours.

The invention provides oocyte culture fluid and a culture method of the oocyte culture fluid. The oocyte culture fluid is easy to prepare and low in cost and comprises G-IVF culture fluid, follicular fluid and granular cells. According to the oocyte culture fluid and the culture method of the oocyte culture fluid, the follicular fluid extracted from the follicle of a patient and granular cells around the oocyte are treated specially and added into the culture fluid, so that in-vitro maturity culturing is carried out on the immature egg of the remaining ICSI period, then fertilization, embryonic development and pregnancy results are analyzed, a simple, convenient and effective in-vitro maturity culture scheme is set up, the effective utilization rate of the immature egg is increased, and more pregnancy opportunities can be provided for the infertility patient.

The invention discloses a culture solution and method for improving the in-vitro maturation quality of porcine oocytes. The culture solution is prepared from the following components: an M199 culturesolution, 10 ng / mL of EGF, 10 [mu]M of asiatic acid, 0.6 mM of L-cysteine, 0.91 mM of sodium pyruvate, 10% follicular fluid, 10 IU / mL of LH, 10 IU / mL of FSH and 100 IU / mL of penicillin. According to the culture solution and method, the asiatic acid is added into the porcine oocyte in-vitro maturation culture solution for the first time, the maturation rate of the oocytes can be effectively increased, the quality of the oocytes can be effectively improved, the development rate of the oocytes can be effectively increased, the quality of the oocytes can be effectively improved, the developmentalrate of parthenogenetic embryos and in-vitro fertilized embryos can be increased, the embryo quality of the parthenogenetic embryos and the in-vitro fertilized embryos can be improved, and the problems of low efficiency and poor quality of in-vitro maturation and in-vitro culture at present are effectively solved.

The invention discloses a porcine ovarian granulosa cell primary culture method. The method comprises steps as follows: ovaries are collected, follicular fluids are sucked, and cells are centrifugally collected; the cells are re-suspended and centrifugally collected to be dispersed, and the process is repeated 2-3 times; the cells are placed in an incubator for culture; solution replacement is performed for the cells after 24 h of culture, a solution, anchorage-independent cells and oocytes are sucked, rinsing is performed once, a fresh solution is added for continuous culture, and the solution is replaced once after 24 h of culture of cells. The method has the benefits as follows: (1) in the granulosa cell collecting process, the step of picking of oocytes with a mouth pipette under a stereomicroscope is omitted, the oocytes suspend to grow in the culture solution, a few of oocytes in a culture dish are sucked away with solution replacement after 24 h of culture, and adherent granulosa cells are pure; (2) the in-vitro operation time is shortened, and the survival rate of the granulosa cells is increased; (3) the obtained granulosa cells is purer.

The invention relates to an egg cells automatic recognition and sorting apparatus, which comprises a negative pressure generator, an egg cell image acquisition device, light source equipment, a follicular fluid collection container, a sorting switch and a central controller which is connected to the negative pressure generator, the egg cell image acquisition device and the sorting switch. The negative pressure generator and the follicular fluid collection container are communicated to a conduit used for sucking and conveying a follicular stock solution with egg cells, and the negative pressure generator is used for forming negative pressure in the tube to complete puncture and suck the follicular stock solution with egg cells into the conduit. The egg cell image acquisition device and the light source equipment are arranged at two sides of a conveying path of the conduit for cooperating to obtain the egg cell image information, the obtained egg cell image information is sent to the central controller, the central controller controls the sorting switch arranged on the conduit to be switched on / off according to the egg cell image information, so that separation of egg cell (a cumulus cell compound) and follicular fluid in the follicular stock solution with egg cells can be realized, and then the required egg cells can be obtained.

As an FAM19A4 protein is possibly related to various physiological and pathological conditions and plays an important role in the physiological and pathological conditions, and quantitative detectionof the soluble FAM19A4 protein in specific samples such as blood, body fluid and cell culture supernatant has significance. The invention firstly relates to an application of the FAM19A4 protein serving as a diagnosis marker of a disease including an infectious disease and an autoimmune disease. The invention further relates to a method and kit for quantitatively detecting the soluble FAM19A4 protein. According to the method and kit, the soluble FAM19A4 protein is detected through a micro-sphere solid carrier and a flow cytometry technique by the aid of the double-antibody sandwich method principle, and the method and kit can be used for quantitatively detecting the FAM19A4 protein in body fluids such as serum, plasma, urine, pleuroperitoneal fluids, joint fluids, cerebrospinal fluids, amniotic fluids and follicular fluids of a clinic patient, basic research and various biological samples such as cell culture fluids and mouse blood in a mouse model.

The invention relates to an oocyte in-vitro maturation culture solution and application thereof, and belongs to the technical field of oocyte in-vitro maturation culture solutions. The oocyte in-vitromaturation culture solution and application thereof are provided for solving the problems that it is necessary to add follicular fluid with unclear compositions into the oocyte in-vitro maturation culture solution, and the culture efficiency is not ideal. The culture solution is composed of TCM-199, KSR, penicillin, streptomycin, NaHCO3, Hepes, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, epidermal growth factors, follicle-generating hormones and luteinizing hormones. According to the oocyte in-vitro maturation culture solution, during oocyte in-vitro culture, the similar effects onthe cleavage rate and the blastocyst formation rate can be obtained as a traditional culture solution with added follicular fluid, the disadvantages that in traditional porcine oocyte in-vitro culture, follicular fluid with unknown components needs to be added can be avoided, and thus an oocyte in-vitro maturation system is more standardized.

The invention relates to a method for improving low ovarian response in patients with polycystic ovary syndrome by transvaginal small follicle puncture. The method comprises case inclusion, measurement indicators, and standard operating procedures; The case inclusion comprises inclusion criteria and exclusion criteria; The measurement indicators comprise main research indicators, follicular fluidanalysis time, secondary research indicators, serum and follicular fluid analysis time, and safety indicators; The standard operating procedures comprises a clinical ovulation promotion protocol and aTVOD method. The advantages of the method of the invention are as follows: the method of COS for improving the low ovarian response in patients with PCOS can be performed on the day after the IVF ovulation promotion cycle immediately, and the effect of COS for improving the low ovarian response in patients with PCOS on the day after the IVF treatment cycle can be evaluated, and the underlying mechanisms can be studied.

The invention discloses a method for improving the developmental capacity of lamb in-vitro embryos and provides a method for accelerating in-vitro maturation of lamb ova. The method comprises the following steps: carrying out in-vitro maturation culture of in-vitro lamb cumulus-oocytes complexes in an ovum maturation accelerating culture solution added with adult lamb follicular fluid, and achieving the in-vitro maturation of the lamb ova. Through the method, the ovum maturation accelerating culture solution added with adult lamb follicular fluid is used for culturing the lamb oocytes; after the ova are mature, the ova can produce in-vitro embryos through in-vitro fertilization; the developmental capacity of the lamb embryos can be obviously improved; the method is simple and feasible and is convenient in material sources.

Disclosed is a method for cloning buffalo's body cells comprising the following steps: charging calf follicular fluid and epithelium cell growth factor into maturation to cultivate acceptor oocyte, enucleating under spindle image system, cultivating and processing donor cells with DNA synthesized depressant Aphidicolin, carrying out hungry cultivation, activating the recombinant embryo with ionomycin and 6-dimethoxy aminopurine, and proceeding embryo extracorporal cultivation with modified TCM 199 and estrus cow's serum.

The invention relates to an oocyte quality evaluation method. The method comprises the step of comparing the composition of follicular fluid metabolites of a subject with low ovarian reserve functionwith that of a healthy subject. The characteristic biomarker is screened through metabonomics analysis, the relationship between the follicular fluid metabolism marker group and the quality of the oocytes is determined, and a more objective and noninvasive oocyte quality evaluation method is established.

The present invention relates to field of fertility. The invention identifies biological ovarian markers from follicular cells, from follicular fluid, from cumulus cells and from oocytes which are indicative of follicular maturity in mammals. Described are methods for improving ovarian stimulation, methods for assessing maturity of a mammalian ovarian follicle, methods for optimizing in vitro maturation (IVM) and methods for classifying an embryo, these methods being based on assessment of expression level of ovarian markers indicative of maturity. Also described are methods for screening compounds stimulatory of or inhibitory to mammalian follicular maturation, kits for evaluating follicular maturity.

The invention provides a method for judging a cell state in a body liquid environment based on a liquid-phase chip technology. A biological marker of a follicular fluid is analyzed by using the liquid-phase chip technology, so as to judge the quality of forming an embryo by an ovum from the same follicular of the follicular fluid; good-quality embryo selection is determined by combining with a morphological analysis. With the adoption of the method provided by the invention, the clinical pregnancy rate and the embryo implantation rate of IVF (In-Vitro Fertilization) can be improved by providing an objective indicator of the ovum quality, so as to improve the result of IVF therapy. A plurality of factor combinations for judging the ovum quality of the follicular fluid can be produced into a commercial liquid-phase chip kit which is used for assisting the reproduction technology.

The invention relates to a method for analyzing the relation between a follicular fluid metabolite and oocyte quality. The method is characterized by comprising a step of comparing the composition ofthe follicular fluid metabolite of a subject with low ovarian reserve with that of the follicular fluid metabolite of a healthy subject. According to the method in the invention, a characteristic biomarker is screened through metabonomics analysis, the relationship between the follicular fluid metabolism marker group and the oocyte quality is determined, and a more objective and non-invasive oocyte quality evaluation method is established.

The invention discloses a collecting device for follicular fluid of an animal in vitro. The collecting device comprises a collecting device body, a flexible pipe, a metal pipe, a buffer solution pipe,a thin plug, an air suction pipe, a rubber plug and a collecting bottle, the collecting device body is composed of a handle, a concave cavity, a rubber ring, an upper cover and several puncturing needles, the upper cover is connected with the flexible pipe, and the flexible pipe is connected with the metal pipe; the buffer solution pipe is composed of an injector adapter located at the upper endof the buffer solution pipe, a middle pipe body and a flushing ball located at the lower end of the buffer solution pipe, and several small holes are formed in the flushing ball; the bottle opening ofthe collecting bottle is plugged by the rubber plug, and the ends of the metal pipe, buffer solution pipe and air suction pipe are all inserted in the rubber plug and appear above the collecting bottle; the other end of the air suction pipe is connected with a vacuum pump, the lower end of the collecting bottle is provided with a glass valve, and a liquid outlet hole is formed in the glass valve.The collecting device for the follicular fluid of the animal in vitro can simply conveniently collect the follicular fluid of the animal in vitro, contamination on a sample cannot be caused during the operative process, and a pipeline can be cleaned conveniently.

The present invention relates generally to the fields of reproductive medicine. More specifically, the present invention relates to in vitro non-invasive methods for determining the quality of an embryo by determining the level of the cell free nucleic acids or miR-29a or let7-b in the nucleic acid extract from a follicular fluid sample.

The invention discloses a method for forming oocytes by induced differentiation of cattle induced pluripotent stem (iPS) cells, comprising three steps of pig follicular fluid preparation, cattle iPS cell culture and the directional differentiation of the oocytes of cattle iPS cells. In the method, the cattle iPS cells are differentiated into oocytes under definite induction condition. As only a small amount of cattle skin cells are inoculated to induce the cattle iPS cells, the application of the cattle iPS cells can effectively avoid the disadvantage that the separation of cattle embryonic stem (ES) cells needs to damage a large amount of excellent embryo. Thus, the method has wide application prospect and development potential in the aspects of cattle disease resistance breeding, mammary bioreactor production, protection and development of excellent genetic resources and the like.

The invention discloses a sheep ovarian granular cell separation, culture and identification method, and belongs to the technical field of animal cell culture. Ovarian granular cells are obtained through special treatment modes of collection, disinfection, cutting and the like. The method has the advantages of being simple, convenient, labor-saving, low in pollution rate, high in follicular fluid obtaining rate and the like, and a foundation is laid for research on development of ovarian granular cells, oocytes and follicles.

Certain compounds, structurally related to natural compounds which can be extracted i.e. from bull testes and from human follicular fluid, can be used for regulating the meiosis in oocytes and in male germ cells.

Certain compounds, structurally related to natural compounds which can be extracted i.a. from bull testes and from human follicular fluid, can be used for regulating the meiosis in oocytes and in male germ cells. Some of these compounds are useful in the treatment of infertility, whereas other compounds are useful as contraceptives. These compounds have the structural formula wherein the substituents are as defined in the specification.

The invention discloses a method for producing dzo gender controllable in vitro embryos. The method comprises the steps of: removing connective tissues surrounding a Bos grunniens ovary and conducting cleaning, selecting oocytes and washing them; transferring the washed oocytes into a culture dish added with serum, hormone, a follicular fluid and a Hepes buffer solution, and then placing the culture dish into an incubator to perform maturation culture; taking X or Y chromosome-containing sperms from Bos taurus, separating dead sperms from live sperms in semen by a Percoll gradient separation liquid; picking the oocytes that are cultured to maturity from the Hepes buffer solution, washing them with a fertilization fluid, then adding the X or Y chromosome-containing sperms into oocyte-containing fertilization microdroplets, and placing the microdroplets into the incubator to conduct in vitro fertilization culture; after fertilization, carrying out a blastocyst maturation treatment; and at a mature time, recovering the blastocyst, and performing embryo transplantation or storage. By combining the method provided in the invention and other modern embryo production technologies, gender controllable dzo breeding can be carried out, and the number of high quality dzos can be increased rapidly.