32results about How to "Improve developmental potential" patented technology

The invention discloses a human ovocyte cryoprotectant. The human ovocyte cryoprotectant contains an ovum refrigerating fluid and an ovum thawing fluid, wherein the ovum refrigerating fluid contains a balance fluid and a refrigerating fluid; the ovum thawing fluid contains a resuscitation fluid, a diluent-1, a diluent-2 and a cleaning solution. The human ovocyte cryoprotectant is developed by combined application of the propanediol and glycol permeable cryoprotectants, and taking the trehalose (trehalose, alpha-D-glucopyranosyl alpha-D-glucopyranoside, C12H22O11) as a non-permeating agent. The non-permeating cryoprotectant suitable for freezing the human ovocyte is developed so that the formation of the ice crystal in the cell can be inhibited, the function of the frozen ovum can be protected and the developmental potentiality of the frozen ovocyte can be greatly improved. The clinical result proves that the fertility rate, the cleavage rate and the high quality blastocyst rate of the frozen ovocyte can be greatly improved.

The invention relates to the technical field of medicines, and provides applications of uterine cavity fluid-derived exosome in preparation of drugs and adjuvant therapeutic agents for treating infertility related diseases. Through the verification of experiments, exosome realizes the treatment or relieving of endometrial growth disorders by promoting the increasing of the thickness of endometrium, enhancing the activity of endometrial cells, reducing the damages to intimal cells, promoting VEGF expression, maintaining the dryness of endometrial stromal cells and stimulating proliferation; andthrough the realizing of the treating or relieving of maternal-fetal immune tolerance disorder-related diseases by achieving immune tolerance effects, treating spontaneous abortion and enhancing T helper lymphocyte levels, a novel theoretical basis can be provided for infertility treatment. In addition, for both fertilized eggs being fertilized in vivo or fertilized in vitro, the exosome can effectively enhance developmental rates in multiples; and implantation rates and farrowing rates of in vitro fertilization transplantation can be increased as well, so that positive auxiliary effects on infertility treatment can be achieved.

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 relates to a computer assisted embryo selection method. Before ICSI is conducted and when short time insemination oocyte dismantling observation is conducted, Nikon NIS-Elements software is used for collecting oocyte images one by one, the thickness of a transparent tape, the diameter of oocytes and the oocyte gap of a polar body position are measured, and the oocytes with the thickness of the transparent tape being 16.9-20.7 microns, the diameter of the oocytes being 114.8-119.9 microns and the oocyte gap of the polar body position being 8.5-12.3 microns are selected for being subjected to insemination; then embryos with the high developmental potentiality are selected. According to the method, the Nikon NIS-Elements software is used for collecting the oocyte images one by one, the thickness of the transparent tape, the diameter of the oocytes and the oocyte gap of the polar body position are measured, the appropriate oocytes are selected for being subjected to insemination, and then the embryos with the high developmental potentiality are selected. According to the method, a layer of screening is added compared with the prior art, and the embryos with the high developmental potentiality can be better obtained. The data obtained by using the Nikon NIS-Elements software for measuring and analyzing are more accurate.

The invention provides a human oocyte in vitro maturation culture solution containing Endothelin-1. The human oocyte in vitro maturation culture solution comprises liquid A and liquid B, wherein the liquid A comprises ET-1, r-FSH and 10% SSS, the liquid B comprises ET-1, r-FSH, hCG and 10% SSS, and solvent is conventional basic culture solutions. Researches show that the culture solution and culture method are capable of promoting human oocyte in vitro maturation especially cytoplasmic maturation and can increase embryo development potential after human oocyte in vitro fertilization. The culture solution aiming at the current situation that development imbalance of cytoplast and cell nucleus during immature human oocyte in vitro culture affects embryo in vitro development potential is developed by utilizing the biological function and effect of ET-1 in cumulus cells. The culture solution has the advantages that the culture solution is applicable to human oocyte in vitro culture, an IVM process can be optimized favorably, and the culture solution is worthy of popularization and application.

The invention relates to a method for preparing an in-vitro oocyte maturation culture solution. The preparation method is characterized by comprising the following steps: 1) preparing a basal IVM (InVitro Maturation) culture medium; and 2) adding lipoprotein into the basal IVM culture medium, thereby obtaining the in-vitro oocyte maturation culture solution. According to the method disclosed by the invention, the problems that the developmental potential of oocyte is poor after IVM and developmental potential of embryos is relatively low after parthenogenetic activation can be solved, the embryo can continuously develop to blastula after the lipoprotein is added, the developmental potential of the embryos is improved by the IVM culture solution added with the lipoprotein, and the embryo damage problem is effectively improved.

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 an oocyte activating physical method and a fertilization potential evaluation index thereof. An electronic pulse sequence is used for activating the oocyte; the calcium oscillation feature formed by calcium ion concentration change in the endochylema detected in a confocal microscopy is used as an oocyte fertilization potential evaluation index.

The invention discloses a human oocyte sequence vitrification refrigerating fluid and thaw liquid belonging to the cell preservation field in biology, which adopts human oviduct fluid which contains HEPES and human blood protein as basic liquid and low-density Ficoll 70, sucrose, ethylene glycol and dimethyl sulphoxide as protective agent. Compared with the prior technique, the human oocyte sequence vitrification refrigerating fluid and thaw liquid of the invention is low in toxicity, good in preservation effect and the like, which can be widely applied in the vitrification preservation of human oocyte.

The invention provides embryo antioxidation related lncRNA and application thereof, and belongs to the technical field of lncRNA function research. A nucleotide sequence of the lncRNA CUFF.97956.1 is as shown in SEQ ID No.1; a target gene of the lncRNA CUFF.97956.1 is a PANX1 gene; the expression quantity of the lncRNA CUFF.97956.1 in the bovine embryo is remarkably reduced after glutathione treatment; the lncRNA CUFF.97956.1 and the target gene thereof are related to the antioxidant and developmental capacity of the embryo; and the lncRNA CUFF.97956.1 and the target gene thereof provide technical support for improving the developmental potential of the in-vitro embryo.

The invention discloses a fructus momordicae extract mogroside V which has a good function of promoting in-vitro maturation of oocyte, and accordingly, an inventor also establishes a relatively complete culture system-a culture solution and a culture method. The culture solution is based on a conventional culture solution, and the fructus momordicae extract mogroside V is added. A research shows that the in-vitro maturation quality of pig ova can be remarkably increased by using the culture solution, an in-vitro maturation rate of the oocyte can be increased from 79.4 percent to 87.4 percent,and a promotion function on ovum developmental potentiality is obtained, so that the in-vitro maturation rate of the oocyte is greatly increased, and the problems that a current maturation efficiencyis low, a culture period is long and large-scale production is not facilitated are solved; by adopting the fructus momordicae extract mogroside V disclosed by the invention, more high-quality oocyte can be provided for somatic cell nuclear transfer, in-vitro fertilization and somatic cell cloning.

The invention discloses a polypeptide FFDP and application thereof in oocyte in-vitro culture. The amino acid sequence of the polypeptide FFDP is as shown in SEQ ID NO. 1. Experiments find that the polypeptide FFDP can be applied to PCOS disease affected oocyte development, and especially applied to the aspect of PCOS oocyte in-vitro maturation culture, or applied to the aspect of improving the meiosis defect during oocyte in-vitro maturation culture.

The invention provides a human oocyte in vitro maturation culture solution containing Endothelin-1. The human oocyte in vitro maturation culture solution comprises liquid A and liquid B, wherein the liquid A comprises ET-1, r-FSH and 10% SSS, the liquid B comprises ET-1, r-FSH, hCG and 10% SSS, and solvent is conventional basic culture solutions. Researches show that the culture solution and culture method are capable of promoting human oocyte in vitro maturation especially cytoplasmic maturation and can increase embryo development potential after human oocyte in vitro fertilization. The culture solution aiming at the current situation that development imbalance of cytoplast and cell nucleus during immature human oocyte in vitro culture affects embryo in vitro development potential is developed by utilizing the biological function and effect of ET-1 in cumulus cells. The culture solution has the advantages that the culture solution is applicable to human oocyte in vitro culture, an IVM process can be optimized favorably, and the culture solution is worthy of popularization and application.

The present invention discloses a selective ultra-early fertilization observation and remedying method for preventing fertilization failure. The selective ultra-early fertilization observation and remedying method comprises that (1) early fertilization observation is performed at 40-60 min after fertilization; (2) at 3-6 layers of cumulus cells closely connected to oocytes, an annular fracture zone is presented or all cumulus cells are scattered; (3) when the light of a microscope passes through the fracture zone, an O-shaped light ring or completely-scattered condition is presented; and (4) the oocytes of the patient having the fertilization ability show the characteristics while the oocytes of the patient with the fertilization failure do not show the characteristics. According to the present invention, with the method, the subject with the fertilization failure can be effectively and accurately found as early as possible, and compared with the conventional method, the ICSI remedying has the following clinical results that the high quality embryo blastula developmental rate is high while the multi-sperm fertilization rate is not increased; and with the method, the fertilization failure can be found as early as possible and the ICSI remedying can be performed as early as possible, the sperm-egg co-incubation time can be reduced, the interference on most of the oocytes having the normal fertilization ability can be avoided, the embryonic development potential can be improved, the live birth rate can be improved, the labor time can be reduced, and the effectiveness of IVF can be improved.

The invention discloses a secreting type endogenous polypeptide PDFF-CO3 and an application thereof. The secreting type endogenous polypeptide PDFF-CO3 is characterized in that an amino acid sequenceof the secreting type endogenous polypeptide PDFF-CO3 is shown as SEQ ID NO. 1. The invention also discloses the application of the secreting type endogenous polypeptide PDFF-CO3 in in-vitro maturation culture of oocytes. The PDFF-CO3 is a CO3 source peptide and is composed of 25 amino acids, and related functions of PDF-CO3 are not reported at present. The PDFF-CO3 is added into a mouse GV-stageegg cell culture solution, so that the biological function of promoting egg cell maturation is achieved; therefore, the PDFF-CO3 can be applied to preparation of medicines for promoting egg cell maturation and improving egg cell development potential and preparation of a culture solution.

The invention discloses a human ovocyte cryoprotectant. The human ovocyte cryoprotectant contains an ovum refrigerating fluid and an ovum thawing fluid, wherein the ovum refrigerating fluid contains a balance fluid and a refrigerating fluid; the ovum thawing fluid contains a resuscitation fluid, a diluent-1, a diluent-2 and a cleaning solution. The human ovocyte cryoprotectant is developed by combined application of the propanediol and glycol permeable cryoprotectants, and taking the trehalose (trehalose, alpha-D-glucopyranosyl alpha-D-glucopyranoside, C12H22O11) as a non-permeating agent. The non-permeating cryoprotectant suitable for freezing the human ovocyte is developed so that the formation of the ice crystal in the cell can be inhibited, the function of the frozen ovum can be protected and the developmental potentiality of the frozen ovocyte can be greatly improved. The clinical result proves that the fertility rate, the cleavage rate and the high quality blastocyst rate of the frozen ovocyte can be greatly improved.

The invention discloses a cryoprotectant and a freezing method for ultra-low temperature storage of copepod eggs. The cryoprotectant comprises trehalose containing L-(-)-trehalose and seawater containing eggs, mainly comprising the following components and parts by weight: 80-110 parts of seawater containing eggs, 1.2-3.8 parts of ethylene glycol , 2.1-4.3 parts of glycerin, 0.6-1.2 parts of dimethyl sulfoxide, and 1.2-2.5 parts of trehalose. The freezing method of the present invention uses the above-mentioned cryoprotectant to carry out multi-stage freezing. The beneficial effects are: the cryoprotectant of the present invention can play a gaining role, can effectively inhibit the formation of ice crystals in egg cells, weaken the damage degree of ice crystals to eggs, improve the hatching rate of copepod eggs, and improve the efficiency of frozen-thawed eggs. developmental potential; the freezing method of the present invention can cooperate with the application of the cryoprotectant to effectively control the osmotic pressure change of the eggs, avoid the formation of intracellular ice and ice, and reduce the ice crystal damage of the copepod eggs.

The invention discloses a fructus momordicae extract mogroside V which has a good function of promoting in-vitro maturation of oocyte, and accordingly, an inventor also establishes a relatively complete culture system-a culture solution and a culture method. The culture solution is based on a conventional culture solution, and the fructus momordicae extract mogroside V is added. A research shows that the in-vitro maturation quality of pig ova can be remarkably increased by using the culture solution, an in-vitro maturation rate of the oocyte can be increased from 79.4 percent to 87.4 percent,and a promotion function on ovum developmental potentiality is obtained, so that the in-vitro maturation rate of the oocyte is greatly increased, and the problems that a current maturation efficiencyis low, a culture period is long and large-scale production is not facilitated are solved; by adopting the fructus momordicae extract mogroside V disclosed by the invention, more high-quality oocyte can be provided for somatic cell nuclear transfer, in-vitro fertilization and somatic cell cloning.

The invention discloses a secreted endogenous polypeptide PDFF‑CO3 and its application. A secreted endogenous polypeptide PDFF‑CO3, characterized in that the amino acid sequence is shown in SEQ ID NO.1. Application of the secreted endogenous polypeptide PDFF-CO3 of the present invention in in vitro maturation and culture of oocytes. PDFF‑CO3 is a CO3-derived peptide, consisting of 25 amino acids, and there is no related function report yet. Adding PDFF‑CO3 to the egg cell culture medium in the GV stage of mice has the biological function of promoting egg cell maturation; therefore, PDFF‑CO3 can be used in the preparation of drugs for promoting egg cell maturation and improving the developmental potential of egg cells, and in the preparation of culture media.

The invention discloses a method for improving the success rate of assisted reproduction and a verification method and system.The method for improving the success rate of assisted reproduction comprises the steps that a plurality of culture solutions obtained after blastocyst culture are obtained, and free DNA of embryos on the sixth day to the seventh day is extracted from the culture solutions; the free DNA is used for carrying out embryo uploidy analysis, uploidy scores of all embryos are compared, priority ranking is carried out according to the uploidy scores of the embryos, and the embryos with high priorities have high development potential; according to the method, endometrial receptivity analysis is conducted through an endometrial sample, the receptivity of the endometrial is judged, a corresponding transplantation strategy is made according to a result, the receptivity comprises a receptivity period and a non-receptivity period, and the corresponding transplantation strategy is that the endometrial in the receptivity period state is in a transplantable state, and the endometrial in the non-receptivity period state is in a non-transplantable state; the transplantation time of the endometrium in the non-receptive state is adjusted to reach a transplantable state. The technical scheme can effectively improve the success rate of assisted reproduction.

The invention provides a collection device and method for uninterruptedly obtaining ova, the device comprises an ova collection tube, a negative pressure compensation tube, a follicular fluid storage tank and a constant temperature incubator, the ova collection tube comprises a tube body with an opening at the bottom and a filter sieve arranged at the lower port of the tube body; an opening is formed in the bottom of a tube body of the negative pressure compensation tube and has the same diameter as the ovum collection tube; the follicular fluid storage tank is hermetically connected with the ovum collection tube and the negative pressure compensation tube; the constant-temperature incubator is a hollow box body which is provided with a heating device and is used for placing the follicular fluid storage tank as well as the ovum collection tube and the negative pressure compensation tube which are connected with the follicular fluid storage tank. According to the invention, uninterrupted ovum collection in the ovum collection operation is achieved, the collection tube does not need to be suspended for replacement, the efficiency of the ovum collection operation is improved, the in-vitro exposure time of ovum is shortened, the pollution risk of the ovum is reduced, the ovum development potential is improved, and subsequent ovum collection and culture operation is facilitated through the matched design of the filter sieve and the sterile culture dish.

The invention discloses a method for unfreezing a gamete or an embryo on a frozen carrier.The method sequentially includes the following steps that a vessel special for vitrification unfreezing is prepared, a vessel disc of the vessel special for vitrification unfreezing is provided with an unfreezing liquid hole with the upper portion being open, the left side wall of the unfreezing liquid hole is an inclined plane, and the bottom of the unfreezing liquid hole is a flat bottom; unfreezing liquid is added into the unfreezing liquid hole; a frozen carrying pole is adopted to take the gamete or the embryo out of liquid nitrogen, the gamete or the embryo is located on a blade of the frozen carrying pole, and the blade is made to downwards move in an inclined mode along the left side wall of the unfreezing liquid hole till the blade is soaked in the unfreezing liquid; a frozen carrying pole is utilized to observe that the gamete or the embryo falls.The method has the advantages that the gamete or the embryo on the frozen carrier can be unfrozen rapidly and accurately, so that developmental potentiality and pregnancy outcome of the gamete or the embryo are improved.The invention further discloses the vessel special for vitrification unfreezing.

The invention discloses a cultivation method for improving the survival rate of black-spotted frogs used in paddy field cultivation, which belongs to the technical field of ecological agriculture cultivation, and includes: S1. Screening before breeding: hatching, cultivating and screening frog eggs in the hatching period; S2. Phase one screening: cultivation and screening of tadpoles in growth phase; S3. Phase two screening: cultivation and screening of tadpoles in metamorphosis phase. Through the multiple screening process of the black-spotted frog seedlings from the incubation period to the metamorphosis period, the young frog seedlings with good development potential, activity ability and adaptability are screened out, which is conducive to improving the survival of the black-spotted frog in rice field culture Rate.

The invention relates to the field of medical technology, and provides the use of exosomes derived from uterine cavity fluid in the preparation of drugs and auxiliary therapeutic agents for infertility-related diseases. It has been confirmed by experiments that exosomes can achieve endometrial growth disorder by promoting the increase of endometrial thickness, enhancing endometrial cell viability, reducing endometrial cell damage, promoting VEGF expression, maintaining endometrial stromal cell stemness and stimulating proliferation. Treatment or remission of diseases; by exerting the role of immune tolerance, treating spontaneous abortion, and increasing the level of T helper lymphocytes, the treatment or remission of diseases related to maternal-fetal immune tolerance disorder is realized, which provides a new method for infertility treatment Theoretical basis. In addition, the exosomes of the present invention can effectively increase the development rate of fertilized eggs fertilized in vivo or fertilized eggs in vitro, and at the same time improve the implantation rate and litter rate of in vitro fertilization rate transplantation , Play a positive auxiliary role in the treatment of infertility.

The invention provides a method for culturing embryonic cells in vitro, and according to the embodiment of the invention, the method comprises the following steps: (i) in vitro culturing fertilized egg cells or derivatives thereof in the presence of an NAD (Nicotinamide Adenine Dinucleotide) signaling pathway agonist. In the culture process of fertilized egg cells or derivatives thereof, the NAD signal channel agonist is added, so that the quality, such as development potential, of embryos cultured in vitro can be effectively improved.

Provided is a fuel oil composition comprising a liquid fuel and carbon black with a particle diameter less than 1 micron. The content of the carbon black is from 0.001 wt % to 4 wt % based on the total weight of the liquid fuel. By way of adding carbon black with the particle diameter less than 1 micron, combustion efficiency of the fuel oil composition used in the internal combustion engine is improved, reducing the emissions of the hydrocarbon and carbon monoxide to reduce greenhouse gas production.