16 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 invention provides a method for determining the gonadal index of oysters, and relates to the field of aquaculture technology. The method for determining the gonadal index of oysters comprises the following steps: S1, dissection: separating the left and right shells of the oysters and taking out the entire soft tissue of the oysters; S2, freezing: after the entire soft tissue of the oysters is dried of surface moisture, it is frozen at -24 ° C for 1-2 hours to set; S3, peeling: peeling off the gonadal tissue wrapped on the digestive caeca, and placing it and the remaining soft tissue after peeling in a fixed container respectively; S4, drying: drying the gonadal tissue and the remaining soft tissue at low temperature for 1-2 hours respectively; S5, weighing: weighing the gonadal tissue and the remaining soft tissue after drying respectively to obtain the gonadal tissue weight W1 and the remaining soft tissue weight W2; S6, formula calculation. The present invention can accurately measure the gonadal index of oysters, solve the problem of low measurement accuracy of traditional methods, and thus more accurately judge and evaluate the fullness and maturity of oyster gonads.

The present application relates to the field of radioactive toxicity detection, and more particularly to a method for evaluating low-activity radioactive 137 Cs toxicity based on marine Oryzias latipes, which comprises the following steps: step 1, setting the breeding conditions of Oryzias latipes, starting the exposure experiment in the adjusted exposure parameters of the radioactive 137 Cs; step 2, observing the reproductive state parameters of Oryzias latipes after the exposure experiment; step 3, dissecting the Oryzias latipes to obtain gonadal tissue; step 4, analyzing and comparing the significant level differences of each index of the control group and the experimental group by using single factor variance statistics of the data information in steps 2 and 3, and determining the toxicity level of the radioactive 137 Cs by the index differences. The present application realizes rapid evaluation of low-activity radioactive 137 Cs toxicity in seawater, and has the characteristics of simple processing process, high operation repeatability and high sensitivity.

The invention discloses a primer and a method for quickly identifying the molecular level gonad differentiation state of strongylocentrotus nudus, which are characterized in that two gene segments CYCLIN A and CYCLIN B which are obviously differentially expressed in undifferentiated and differentiated gonad tissues are analyzed and identified through bioinformatics, a fluorescent quantitative PCR (Polymerase Chain Reaction) primer is designed, and a Ubiquitin gene is used as a reference gene through real-time quantitative PCR, so that the differentiated gonad differentiation state of strongylocentrotus nudus is identified, and the differentiated gonad differentiation state of strongylocentrotus nudus is identified. Respectively calculating expression quantities of the CYCLIN A gene and the CYCLIN B gene in the same sea urchin gonad, and converting the expression quantities into-LOG2 multiple change values; if the obtained-LOG2 multiple change values are all greater than 3, determining that differentiation is not carried out, otherwise, determining that differentiation is carried out. Compared with the prior art, the method has the characteristics of simplicity, rapidness, accuracy, sensitivity and the like.

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 invention relates to the field of aquatic animal physiology research, and discloses a method for detecting the oxidative stress level of stichopus japonicus in the breeding period based on mitochondrial membrane potential, which adopts a flow cytometry detection method to detect the mitochondrial membrane potential formed in gonad tissue or coelomic fluid in the breeding process of the stichopus japonicus as an index for detecting the oxidative stress level. The method takes the mitochondrial membrane potential as a detection index, can detect the oxidative stress level of the stichopus japonicus more accurately and quickly, focuses on gonad tissue and coelomic fluid of the stichopus japonicus, and can provide guarantee for healthy development of the stichopus japonicus industry. The method provided by the invention provides an innovative technical means for existing research, breaks through the technical bottleneck of sea cucumber germ cell detection, establishes a standardized flow type detection system of the stichopus japonicus gonad tissue for the first time, and fills the methodology blank of stichopus japonicus reproduction oxidative stress research so as to guarantee the comprehensiveness of stichopus japonicus physiological research; and a technical guarantee is provided for researchers to monitor the breeding and growth data of the stichopus japonicus.

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.

The invention relates to the field of aquatic animal physiology research, and discloses an ROS-based method for detecting the oxidative stress level of stichopus japonicus in a breeding period, and ROS generated in gonad tissue or coelomic fluid in the breeding process of the stichopus japonicus is detected by adopting a flow cytometry detection method as an index for detecting the oxidative stress level. The ROS is directly used as a detection index, the oxidative stress level of the stichopus japonicus can be detected more accurately and quickly, gonad tissue and coelomic fluid of the stichopus japonicus are focused, and a guarantee can be provided for healthy development of the stichopus japonicus industry. The method provided by the invention provides an innovative technical means for existing research, breaks through the technical bottleneck of ROS detection of the germ cells of the sea cucumbers, establishes a standardized flow type detection system of the gonad tissues of the stichopus japonicus for the first time, fills the methodology blank of research on reproductive oxidative stress of the stichopus japonicus so as to guarantee comprehensiveness of physiological research of the stichopus japonicus, and has a wide application prospect. And a technical guarantee is provided for researchers to monitor the breeding and growth data of the stichopus japonicus.