173 results about "Ovarian tissue" patented technology

Methods and compositions for identifying ovarian cancer in a patient sample are provided. The methods of the invention comprise detecting overexpression or underexpression of at least one nucleic acid biomarker in a body sample, wherein the biomarker is selectively overexpressed or underexpressed in ovarian cancer. The body sample may be, for example, an ovarian tissue sample. The biomarkers of the invention include any nucleic acid molecule that is selectively overexpressed in ovarian cancer, including, for example, MMP-7, PAEP, CA125, HE4, PLAUR, MUC-1, SLPI, SSP1, MSLN, SPON1, interleukin-7, folate receptor 1, claudin 3, inhibin A, inhibin BB, inhibin BA, and PAI-1. Overexpression or underexpression of a biomarker of interest is detected at the nucleic acid level using such methods as real-time PCR and various nucleic acid hybridization techniques. Kits for practicing the methods of the invention are further provided.

This invention provides VEGF-FSH compounds having increased serum half-lives relative to either native VEGF or FSH, in which both VEGF and FSH are biologically active. This invention also provides related compositions and methods for increasing fertility, egg production and spermatogenesis in a subject, as well as methods for increasing vascularization in a tissue, particularly in ovarian tissue.

The invention discloses a natural tissue derived ovarian acellular material and a preparation method thereof. The acellular ovarian material is prepared by processing any ovarian tissues of a mammal by virtue of a physiological saline buffer solution containing a protease inhibitor, an organic solvent solution, a PBS buffer solution containing Triton X, a PBS buffer solution containing SDS and a PBS buffer solution containing DNA enzyme. According to the ovarian acellular material provided by the invention, allogeneic or xenogeneic cells, which have immunogenicity, are removed, and meanwhile, original ECM integrity can be reserved; the ovarian acellular material has excellent extracellular microenvironment, biochemical factors, biomechanical properties and the like; and complex tissue structure of a normal ovary can be simulated to the greatest extent.

The invention discloses a large ovarian tissue vitrification freezing carrier. The carrier consists of a carrier inner core and an outer sleeve, wherein the carrier inner core is in threaded connection with the outer sleeve, maintaining high airtightness and preventing cross infection of the ovarian tissue; meanwhile, the user operation is also facilitated; the carrier inner core consists of a handheld part, a cap part, a support part and a bearing part sequentially from top to bottom; the bearing part is clamped and fixed by the support part; when in use, a large ovarian tissue is put on an end part of the bearing part, away from the support part; the end part is a large ovarian tissue loading area; experiments prove that by using the carrier disclosed by the invention, the permeation efficiency of a refrigerant is improved; moreover, the operation is simple, the price is low, batch storage of large ovarian tissues is convenient, and the carrier is suitable for promotion and application.

The extracorporeal culture and mature method of immature oocyte from ovary tissue block includes selecting marrow mesenchyme stem cell MSC of female mouse as the feeder cell, compounding MSC culture liquid and agar and covering the culture rack, adding HCG in 3 IU / ml, and adhering the ovary tissue block on the rack to culture. In the co-culture system, estradiol, progestin and luteotopin contents are increased obviously, and after culturing, there is mature oocyte discharged. The method is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

A method for aspirating an oocyte from a human female is provided. The method includes the step of providing at least one ovarian cortical piece, implanting the ovarian cortical piece in the human female at a heterotopic location, and triggering oocyte maturity in the at least one ovarian cortical piece. The method also includes the steps of providing an aspiration needle and retrieving at least one oocyte from the at least one ovarian cortical piece using the aspiration needle.

The present invention relates to newly identified nucleic acid molecules and polypeptides present in normal and neoplastic ovarian cells, including fragments, variants and derivatives of the nucleic acids and polypeptides. The present invention also relates to antibodies to the polypeptides of the invention, as well as agonists and antagonists of the polypeptides of the invention. The invention also relates to compositions containing the nucleic acid molecules, polypeptides, antibodies, agonists and antagonists of the invention and methods for the use of these compositions. These uses include identifying, diagnosing, monitoring, staging, imaging and treating ovarian cancer and non-cancerous disease states in ovarian, identifying ovarian tissue, monitoring and identifying and / or designing agonists and antagonists of polypeptides of the invention. The uses also include gene therapy, production of transgenic animals and cells, and production of engineered ovarian tissue for treatment and research.

The invention belongs to the field related to biological tissue or cell preservation, and relates to a novel tissue cryopreservation liquid, in particular to a liquid used for cryopreserving tumor tissue or ovarian tissue. The cryopreservation liquid is prepared from a Leibovitz L-15 culture medium, fetal calf serum and 14-18% of DMSO (dimethyl sulfoxide), wherein the Leibovitz L-15 culture medium (Gibco) can be purchased or can be self-made by technicians in the field according to a reported formula; preferably, the content of DMSO is 14-16%, more preferably, 15%. For the novel cryopreservation liquid, preferably, the content of fetal calf serum is 14-18%, more preferably, 14-16%, the most preferably, 15%. The cryopreservation liquid has the advantages of being long in cryopreservation time, high in tumor formation rate and the like, the preparation method is simple, and the raw material source is wide.

The present invention relates to newly identified nucleic acid molecules and polypeptides present in normal and neoplastic ovarian cells, including fragments, variants and derivatives of the nucleic acids and polypeptides. The present invention also relates to antibodies to the polypeptides of the invention, as well as agonists and antagonists of the polypeptides of the invention. The invention also relates to compositions containing the nucleic acid molecules, polypeptides, antibodies, agonists and antagonists of the invention and methods for the use of these compositions. These uses include identifying, diagnosing, monitoring, staging, imaging and treating ovarian cancer and non-cancerous disease states in ovarian, identifying ovarian tissue, monitoring and identifying and / or designing agonists and antagonists of polypeptides of the invention. The uses also include gene therapy, production of transgenic animals and cells, and production of engineered ovarian tissue for treatment and research.

The method of obtaining immature oocyte from ovary tissue through separating or extracting for extracorporeal culture includes selecting marrow mesenchyme stem cell MSC of female mouse as the feeder cell or culturing the immature oocyte in the MSC culture liquid. After culturing, the co-culture system has obviously increased estradiol, progestin and luteotopin contents, and over 90 % of the immature oocyte become mature. The method is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

The invention provides a method for knocking out a CRISPR / Cas9 mediated sheep FGF5 gene and integrating an MTNR1A gene at a fixed point.The method includes the steps of transferring the CRISPR / Cas9 targeting vector of the specific targeting third exon of the sheep FGF5 gene and the gene homologous recombination vector into a sheep fetus fibroblast to obtain a sheep FGF5 gene knockout cell with a fixed-point integration of the MTNR1A gene. The donor vector and the targeting vector are co-transfected into sheep fetus fibroblasts in a nuclear transfer mode, and specific expression of MTNR1A in sheep ovary tissue can be achieved through a CYP17 promoter in the unmarked donor vector constructed through a Gibson Assembly method. The separated and identified positive monoclonal cell strain can beused as a nuclear donor for nuclear transplantation of somatic cells to obtain cloned embryos, so that a solid foundation is laid for culturing transgenic clone sheep which locally strengthens melatonin signals and produces more hairs in a reproductive system.

The invention aims at providing an establishment method of an ovary cell line of cynoglossus semilaevis, which comprises the following steps: 1) removing the outermost membrane of the ovary tissue of cynoglossus semilaevis under a sterile condition, cutting the tissue into small blocks in a culture solution, flushing the tissue blocks with a PBS solution and digesting with a trypsin solution, wherein the solution after trypsin digestion contains cells, cell clusters and tissue blocks not completely digested; and 2) centrifuging the solution to remove the supernatant trypsin, and suspending the cells, cell clusters and small tissue block precipitates with a culture solution; after a suspension is obtained, inoculating into a culture plate treated by the type-I collagen; adding a culture solution, and culturing in a biochemical incubator at 24 DEG C; and performing subculture to finish establishment of the ovary cell line. In the establishment method provided by the invention, an ovary cell line of cynoglossus semilaevis is successfully established by use of the ovary tissue of cynoglossus semilaevis, the established cell line can be continuously passed, the passage cells obtained by the method can be passed for over 60 generations, and a great quantity of ovary cells can be provided.

The invention relates to a FFRC strain common carp mature ovarian tissue frozen section making method and belongs to the technical field of tissue sections. Mature ovarian tissues are taken out from a FFRC strain common carp, and a FFRC strain common carp mature ovarian tissue frozen section is obtained after fixation, degreasing, dehydration, embedding, freezing and slicing are performed. Compared with a common general freezing and slicing method, degreasing is performed through pure acetone, excessive fat in a later development period of FFRC strain common carp mature ovaries can be efficiently removed, and accordingly the frozen ovarian section high in quality and clearer in structure is obtained. An important and reliable molecular experiment material is provided for tissue localization and functional study such as immunohistochemistry of FFRC strain common carp mature ovarian genes, and the FFRC strain common carp mature ovarian tissue frozen section making method has the important significance on molecular mechanism study of good economic characters of FFRC strain common carps.

The invention discloses a preparation method for a procambarus clarkii ovarian tissue total protein sample suitable for dimensional electrophoresis. The method comprises the following steps: fully grinding ovarian tissue by using the liquid nitrogen method; precipitating proteins by using the trichloroacetic acid-acetone method, that is, precipitating the proteins by using a precooled acetone solution containing dithiothreitol and trichloroacetic acid and rinsing precipitate with a precooled acetone solution containing dithiothreitol a plurality of times; adding a sample lysate to prepare the high quality procambarus clarkii ovarian tissue total protein sample suitable for dimensional electrophoresis. The preparation method for the sample suitable for dimensional electrophoresis is rapid, reliable, simple and practicable, enables a dimensional electrophoresis pattern with high quality and good experimental repeatability to be obtained and subsequent biological mass spectrometric analysis and identification of the proteins to be convenient, and has a great practical application value.

The invention relates to the field of reproductive medicines, in particular to an ovarian tissue cryopreservation method. The ovarian tissue cryopreservation method comprises the following steps: culturing an ovarian tissue to be frozen by a hyaluronidase solution with the volume concentration of 8-12 percent for 10-15 minutes so as to obtain an enzymolyzed ovarian tissue; adding the enzymolyzed ovarian tissue into a freezing protection agent for prebalancing under the temperature of 3-5 DEG C for 10-15 minutes to obtain a prebalanced ovarian tissue; performing programmed cryopreservation on the prebalanced ovarian tissue to reduce the temperature of the prebalanced ovarian tissue to be minus 150 DEG C, and transferring the prebalanced ovarian tissue into a liquid nitrogen tank for storage. Compared with the common prebalancing time of 30-90 minutes in the prior art, the prebalancing time of the ovarian tissue cryopreservation method is greatly shortened, so that the cytotoxic damage, which is caused by the freezing protection agent, to the ovarian tissue under overlong prebalancing time is avoided.

The described invention provides methods for detecting, diagnosing and treating low-grade ovarian cancer and stage I ovarian cancer by comparing results from serum and ovarian tissue samples with normal controls. An increased level of expression of serine protease, wherein the serine protease is at least 2 selected from the group consisting of kallikrein 6 (KLK6), kallikrein 7 (KLK7), and PRSS8, expressed by subject samples compared to the level of expression of serine protease expressed by normal control samples is indicative of possible early stage ovarian cancer in the subject. Once early stage (I / II) ovarian cancer is diagnosed, the subject is treated with a treatment regimen effective to treat the early stage (I / II) ovarian cancer.

A molecular diagnosis system of ovarian cancers encompasses a detection device configured to obtain a detected value of an expression amount of an apolipoprotein A1 gene in ovarian tissue as a diagnosis subject, a storage device configured to store a normal value of the expression amount of the apolipoprotein A1 gene in normal ovarian tissue, and a determination mechanism configured to determine that the ovarian tissue as the diagnosis subject is clear cell adenocarcinoma when the detected value is lower than the normal value.

The present invention is feeder cell for extracorporeal culture of immature oocyte and features that marrow mesenchyme stem cell MSC of female mouse is used as the feeder cell. The feeder cell is easy to obtain, can secrete several kinds of cell factors and growth factors, promote over 90 % of the immature oocyte to become mature and promote the growth, ovulation and maturing of the ovarian follicles in the ovary tissue block under extracorporeal culture. The present invention is significant in surviving rear endangered species of animals, breeding fine variety of animal, cell biological research of embryo stem cell, etc.

The invention discloses a method for quickly and effectively transfecting mosquito by using wolbachia. According to the method, an insects ovarian tissue (abundant sources and easiness for obtaining) containing the wolbachia is directly acquired by adopting a vivisection method and is homogenated; and the ovarian tissue is separated from the wolbachia by using a centrifugation method so as to obtain a purified usable wolbachia transfected by embryonic injection. The embryo of the mosquito is injected and transfected by the purified wolbachia through a conventional method; a primary female adult insect (G0) is obtained by egg culture; the obtained primary female adult insect is mated with a wild type male mosquito to generate first generation (G1) mosquito strains; and positive mosquito strains containing the wolbachia are screened from the first generation mosquito strains and become really and successfully-transfected mosquito strains. Therefore, the method for transfecting the mosquito by using the wolbachia is a quick and effective method. The realization of the invention is beneficial to quickly establishing the mosquito strains transfected with the wolbachia and further beneficial to large-scale application of the mosquito strains.

The present invention relates to a tumor marker for diagnosis of ovarian cancer, which is selected from the group consisting of galectin-1, cathepsin B, MHC class I antigen, heat shock protein (HSP) 27, ubiquitin carboxy-termal esterase L1, plasma retinol-binding protein (PRBP), transthyretin, SH3 binding glutamate-rich protein, tubulin-specific chaperone A, RNA binding protein regulatory subunit, γ-actin, tropomyosin and calcium / calmodulin-stimulated cyclic nucleotide phosphatase. The ovarian cancer is diagnosed effectively and efficiently based on detecting the expression levels of the tumor markers in the invention from the ovarian tissue sample of an individual to be diagnosed.

An apparatus and method of use thereof for identifying and monitoring women at risk of developing OSE-derived carcinomas is provided. The apparatus includes an introducer needle configured to be capable of insertion into a female such that a terminal end of the needle is positioned adjacent an ovary of the female, a microendoscope having an optic fiber which is operably insertable into the needle in a manner to enable an image of the ovary to be obtained therethrough, and a tissue removing member operably co-insertable into the needle with the optic fiber therein to enable removal of ovarian tissue cells with minimal deleterious effect to the ovary.

A method and device for freeze-drying a biological sample of mammalian cells or tissue are disclosed. The method includes placing a biological sample on or in a structure to increase a temperature ofthe biological sample and with the biological sample in a closed chamber applying a vacuum to the chamber to lower a pressure within the chamber, cooling the chamber to lower a temperature within thechamber and applying heat to the biological sample within the chamber. The biological sample can include one or more of stem cells, hematopoietic stem cells, mesenchymal stem cells, embryonic stem cells, induced pluripotent stem cells, sperm, oocytes, embryos, ovarian tissue, uterine tissue or testicular tissue. A method for rehydration of a sample is further disclosed.

The invention relates to an application of a fingerprint spectrum consisting of microRNAs in the diagnosis and treatment of human ovarian cancer. The fingerprint spectrum consisting of multiple microRNAs can be used for distinguishing ovarian tissue from ovarian cancer tissue very effectively, has high sensitivity and specificity and can be effectively applied to the ovarian cancer diagnosis, tumor grading and prognosis evaluation.

The invention relates to therapeutic and prophylactic treatment of ovarian cancer and metastases thereof. More specifically, the invention relates to immunogenic polypeptides comprising at least a portion of an ovarian tissue cell-associated protein or immunologically active variants thereof and to nucleic acids encoding such polypeptides and to the use thereof in immunotherapeutic methods of treatment. Said immunogenic polypeptides are provided by the zona pellucida (ZP) glycoproteins. ZP glycoproteins and fragments thereof that can induce a CD8+ and / or CD4+ T cell response as well as nucleic acid sequences encoding them can suitably be used in the present immunotherapeutic strategies.

The invention discloses a culture medium for 3D culture of ovarian cancer tissue, the tissue is prepared from cytokine B27, N-acetylcysteine, R-spondin 1, A83-01, epidermal growth factors, glutamine,N2Supplement, estrogen, progesterone and vitamin E. The culture medium includes various cytokines and signal pathway regulatory factors which are needed by ovarian cancer tissue cell culture, mutually, directly and closely affect each other and coordinate with each other, so that the ovarian cancer tissue cells can better display the inherent active characteristics in the process of culture, and the overall characteristic highly similar to the living ovarian cancer tissue is achieved, and ovarian tumor cells subjected to 3D culture with the culture medium are clustered and are hypoxic in the middle and similar to the ovarian tumor tissue.

A device for freeze preservation of a human ovarian tissue under liquid nitrogen comprises a pipe body; one end of the pipe body is provided with an opening, the opening is equipped with a sealing cover, the sealing cover and the pipe body can be detachably connected, and a sealed storage space is enclosed by the sealing cover and the pipe body; the inner top surface of the sealing cover is provided with a forking device for forking the human ovarian tissue. The forking device comprises a plurality of fine needles which are arranged in parallel, and fixed ends of the fine needles are vertically fixed on the inner end surface of the sealing cover, free ends of the fine needles extend to the pipe body along the axial direction of the pipe body. The forking device comprises 3 fine needles, the 3 fixed ends of the 3 fine needles are in head-to-tail connection to form a regular triangle, and the circle center of the inner end face of the sealing cover is the center of the regular triangle. The device is simple in structure and easy to use; the forking device can avoid physical damage of blind clipping with forceps under liquid nitrogen on an ovarian follicle enriched cortical part in the ovarian tissue, and also avoids cross contamination with other iatrogenic substances in a liquid nitrogen tank.

This invention provides VEGF-FSH compounds having increased serum half-lives relative to either native VEGF or FSH, in which both VEGF and FSH are biologically active. This invention also provides related compositions and methods for increasing fertility, egg production and spermatogenesis in a subject, as well as methods for increasing vascularization in a tissue, particularly in ovarian tissue.

The invention provides a pet ovarian tissue cryopreservation resuscitation reagent which comprises: (1) a cryopreservation resuscitation reagent 1 including 2-7W / V% of lycium barbarum polysaccharide, 6-14V / V% of dimethyl sulfoxide, 2-7V / V% of propylene glycol and 80-90V / V% of an MEM (Memminimum Essential Medium) culture medium; (2) a cryopreservation resuscitation reagent 2 including 8-10W / V% of lycium barbarum polysaccharide, 6-20V / V% of dimethyl sulfoxide, 5-15V / V% of propylene glycol and 70-80V / V% of an MEM culture medium; (3) a cryopreservation resuscitation reagent 3 including 15-20W / V% of lycium barbarum polysaccharide, 6-20V / V% of dimethyl sulfoxide, 5-15V / V% of propylene glycol and 70-80V / V% of an MEM culture medium. The invention further provides a pet ovarian tissue cryopreservation resuscitation reagent group and a pet ovarian tissue cryopreservation and resuscitation method.

The invention discloses a method for screening a miR-181b target gene. The method comprises the following steps: taking a goat ovarian tissue cDNA (complementary Deoxyribonucleic Acid) as a template and carrying out amplification in the presence of Taq DNA polymerase, a buffering environment, Mg<2+> and dNTPs by utilizing a primer RUNX1 under the condition of PCR (Polymerase Chain Reaction), so asto obtain a determined PCR product which is a sequence of a 3' UTR region of an RUNX1 gene; then constructing a dual-luciferase reporting system and detecting the activity of luciferase; primarily identifying the miR-181b target gene; detecting the influences, caused by the fact that miR-181b is detected by adopting an RT-qPCR method, on the level of an RUNX1 gene on mRNA (massager Ribonucleic Acid); detecting the influences, caused by the fact that the miR-181b is detected by adopting a Western blot method, on a protein level of the RUNX1 gene. A combination site with the miR-181b exists inthe RUNX1 3' UTR region; a modern molecular biotechnology is used for verifying a targeting regulation relation of the miR-181b and the target gene RUNX1 and the miR-181b can be used for inhibiting the expression of the RUNX1 gene in the mRNA and protein levels; furthermore, the method confirms that the RUNX1 is a target gent of the miR-181b. The method lays a foundation for further researching influences, caused by the miR-181b, on ovarian development and lambing performance of dairy goats.

The invention discloses an application of a cryoprotectant in organ and tissue cryopreservation. The cryoprotectant contains PVA, ethylene glycol and sucrose. PVA is taken as a main cryoprotectant component of the cryoprotectant, the DMSO content and toxicity are low, and the cryoprotectant has high preservation efficiency when being applied to ovarian tissues or organs. The cryoprotectant is simple in composition and can be widely applied to cryopreservation of various cells, tissues and organs; the raw materials are easily available, and the cost is low.