11 results about "Gonad tissue" patented technology

The invention relates to the technical field of aquatic animal cell culture, in particular to a rapid enrichment and separation method for gonad somatic cells of plectropomus leopardus. Comprising the following steps: selecting healthy plectropomus leopardus for anesthesia; the method comprises the following steps: taking gonad tissues, performing sterile treatment, cutting, centrifugal washing and filtering, inoculating the gonad tissues into a culture medium for primary culture to obtain gonad cells emigrated from tissue blocks, performing trypsin-EDTA digestion and centrifugal washing, and treating the gonad cells with 1 * PBS containing polysorbate; after closing, antibody incubation and washing, the marking effect is verified through a fluorescence microscope, and specific enrichment of gonad somatic cells is achieved; and filtering and purifying the enriched fluorescence-labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescence-positive cells under a fluorescence visual field, transferring the fluorescence-positive cells into a culture dish containing a complete medium, inoculating the separated fluorescence-positive cells into the optimized complete medium, and carrying out purification culture, so that the cells grow in a wall-attached manner and stable passage is realized.

The present application relates to the technical field of aquatic animal cell culture, and particularly relates to a rapid enrichment and separation method of Plectrypops leopardus gonadal somatic cells. The method comprises the following steps: selecting healthy Plectrypops leopardus for anesthesia; taking gonadal tissue, and after aseptic treatment, cutting, centrifugal washing and filtration, inoculating in a culture medium for primary culture to obtain gonadal cells migrated from the tissue block, and after trypsin-EDTA digestion and centrifugal washing, using 1xPBS containing polysorbate for treatment; after blocking, antibody incubation and washing, verifying the labeling effect by a fluorescence microscope to realize specific enrichment of gonadal somatic cells; after filtration and purification of the enriched fluorescent labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescent positive cells under a fluorescence field, and transferring the cells to a culture dish containing complete culture medium, inoculating the separated fluorescent positive cells in optimized complete culture medium for purification culture, so that the cells adhere to the wall and realize stable subculture.

The invention discloses application of H3K9me2 as a target spot in preparation of a preparation for inducing stem cells to differentiate to gametes in vitro, and relates to the technical field of stem cell breeding in animal husbandry. An H3K9me2 inhibitor BIX01294 is adopted as a treating agent, and in-vivo treatment is performed on a pregnant mouse and in-vitro treatment is performed on gonad tissue of a female fetal mouse. The result shows that the BIX01294 treatment can cause the reduction of the expression level of H3K9me2, so that the expression levels of pluripotent genes Oct4, Sox2 and Dppa3 are increased, the accessibility of whole genome chromatin is obviously changed, and the meiosis start and meiosis process are blocked. The result shows that the H3K9me2 can influence the differentiation of the stem cells to the gametes and is beneficial to the generation of artificial gametes in the stem cell breeding process.

The invention provides a myxocyprinus asiaticus individual genetic resurgence method which comprises the following steps: S1, taking a myxocyprinus asiaticus gonad tissue, cleaning, cutting into pieces, adding an L-15 culture medium containing double antibodies and a DPBS buffer solution, and centrifuging to obtain the gonad tissue; s2, performing water bath on the gonad tissues to obtain resuscitated gonad tissues; s3, digesting, filtering and centrifugally purifying the resuscitated gonad tissues to obtain germline stem cells; s4, injecting morpholino to obtain gobiocypris rarus without primordial germ cells; and S5, transplanting the germline stem cells into gobiocypris rarus juvenile fish bodies, and culturing to obtain the myxocyprinus asiaticus gametes. S6, the generated myxocyprinus asiaticus gametes are fertilized, and myxocyprinus asiaticus individuals are obtained. According to the method, the sperms of the myxocyprinus asiaticus are successfully obtained by utilizing a gonad cryopreservation and abdomen borrowing reproduction technology, and a feasible way is provided for individual genetic resurgence of rare and endangered fishes.

The present invention relates to a method, to a computer program product and to a system for detecting gonad maturity in adult mussels. The method comprises the following steps: a) collecting adult mussels from an area of interest; b) extracting fresh gonad tissue samples; c) preparing smears; d) capturing images of the smears with a portable microscope and a digital camera; e) analysing the images using an application on a smart device, which transmits said images to a server equipped with a convolutional neural network model, in order to classify the samples as female, male or undetermined, and predict the degree of maturity in the females; and f) receiving, on the smart device, the results of the sex classification and the degree of reproductive maturity in the analysed samples. The invention further discloses a method for capturing mussel spat that comprises detecting gonad maturity in adult mussels.

The invention relates to the technical field of information, and particularly provides a fish germ cell subcellular structure separation method, which comprises the following steps: obtaining fish gonad tissues in an environment without RNA enzyme pollution, washing with a precooling buffer solution, directly quick-freezing, embedding in an embedding medium, and storing at low temperature; freezing and slicing the quick-frozen and embedded sample to obtain a thin-layer slice, and attaching the thin-layer slice to a membrane subjected to adhesion enhancement treatment and enzyme-free treatment; the attached section is subjected to visual treatment by adopting a specific dyeing combination and then is stored at low temperature; the visualized diaphragm is placed in a humidity-controlled windless enzyme-free environment, and a target membraneless subcellular structure is recognized by combining phase difference or differential interference contrast imaging and three-dimensional stack scanning through a multiple objective lens; the laser microdissection system adopts laser parameters optimized for a small-area target to complete cutting; and adsorbing the cut membraneless subcellular structure into a tube cover of a collecting tube, immediately adding a lysis solution, mixing, and storing at low temperature for nucleic acid extraction.

The invention belongs to the technical field of in-vitro culture of germline stem cells, and particularly relates to a separation and in-vitro culture method of spermatogonial stem cells of grass carp. The method comprises the following steps: by taking male grass carps as donors, anesthetizing, killing, cleaning, disinfecting, dissecting and separating gonad tissues, crushing into fragments by adopting a mode of combining magnetic bead crushing with shearing, digesting with trypsin, filtering and centrifuging to obtain high-purity spermatogonial stem cells, and performing in-vitro culture on the spermatogonial stem cells by adopting a culture solution. By optimizing the age of the male grass carp, an enzyme digestion process and an in-vitro culture system and adopting a one-step enzyme digestion method, the digestion time is shortened, the operation process is simplified, and the industrial problems that the spermatogonial stem cells of the grass carp are low in separation efficiency and culture is easy to differentiate are solved. According to the invention, a separation, purification and primary culture technical system of the spermatogonial stem cells of the grass carp is systematically established for the first time, and important cell materials and technical supports are provided for breeding technologies such as transplantation of germline stem cells of the grass carp, gene editing and the like.

The invention discloses a composition for improving male sexual function and resisting fatigue as well as a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine compositions. On one hand, the composition provided by the invention can promote synthesis and secretion of testosterone and maintain a normal testosterone level of an organism; on the other hand, blood perfusion of gonad tissues is improved, the mitochondrial function of gonad cells is enhanced, testis aging is delayed, and then the effects of enhancing sexual desire and erection functions and improving premature ejaculation and spermatorrhea are achieved; the traditional Chinese medicine composition also has the effects of protecting nerves and cell tissues from oxidative damage and repairing the damaged tissues, so that the penis sensitivity is reduced, and the problem of premature ejaculation is effectively improved or cured; by providing sufficient kidney essence and qi and blood for the body, sleep and psychological disorder / pressure are improved, and the anti-fatigue ability is improved.

The application provides a method for genetic reactivation of individual Procyprinus palpebrosus, comprising the following steps: S1, taking the gonadal tissue of Procyprinus palpebrosus, washing, cutting, adding L-15 culture medium containing double antibodies and DPBS buffer solution to centrifuge the gonadal tissue; S2, water-bathing the gonadal tissue to obtain recovered gonadal tissue; S3, digesting the recovered gonadal tissue, filtering, centrifuging and purifying to obtain germ stem cells; S4, injecting morpholino to obtain rare gobi-ocypris rarus with deleted primordial germ cells; S5, transplanting the germ stem cells into the body of the rare gobi-ocypris rarus juvenile to culture, and obtaining the gametes of Procyprinus palpebrosus; and S6, fertilizing the generated gametes of Procyprinus palpebrosus to obtain the individual Procyprinus palpebrosus. The application successfully obtains the sperm of Procyprinus palpebrosus by using the cryopreserved gonadal tissue and the technology of parthenogenesis, and provides a feasible way for the individual genetic reactivation of rare and endangered fish species.

The invention provides an RNA (Ribonucleic Acid) molecular marker for judging early gonad differentiation of young river snails in China and an identification method. Dmrt1 and Foxl2 genes are important genes related to gonad differentiation of river snails. The method comprises the following steps: designing and verifying primers SEQ ID NO: 1-2 according to conserved regions of Dmrt1 and Foxl2 genes; the method comprises the following steps: collecting gonad tissue samples in a young snail stage and a sexual maturity stage, wherein n is greater than or equal to 120, extracting total RNA (Ribonucleic Acid) of a field snail tissue, carrying out reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid), detecting a double-gene expression quantity through qRT-PCR (Quantitative Reverse Transcription-Polymerase Chain Reaction), calculating a standardized expression value by taking beta-actin as an internal reference, and establishing a discrimination function D = log2 (Dmrt1 / Foxl2); 1, judging that the juvenile snails of the field snails start gonad differentiation; according to the invention, the problems of deficiency of specific molecular markers for sex differentiation of the young escargots in China and indefinite gonad differentiation time point are solved in a breakthrough manner, accurate locking of the gonad differentiation time of the young escargots is realized, the gonad differentiation time is 15-20 days earlier than morphological differentiation, and verification of more than one hundred of samples shows that the identification accuracy reaches 100%.

The invention belongs to the technical field of oyster phenotype analysis, particularly relates to an oyster gonad development level high-throughput accurate phenotype analysis method based on U-Net deep learning, and effectively solves the problems of complicated operation, high subjectivity and low precision and throughput in existing oyster gonad development phenotype identification. The method comprises the following steps: acquiring an oyster gonad tissue slice image, constructing and training a U-Net deep learning segmentation model, automatically identifying and segmenting a gonad tissue region in the image, further accurately calculating the Gonadal Proportion (GP) of the gonad tissue in the total gonad area, and realizing high-throughput and accurate quantification of the gonad development level. According to the method, the gonad tissue and the connective tissue can be accurately distinguished, objective phenotypic data are provided, key technical support is provided for gonad development character genetic parameter estimation and molecular assisted breeding of large-scale groups, and the efficiency, accuracy and practicability of gonad development phenotypic analysis are improved.