105 results about "Trophoblast cell" patented technology

The invention provides an embryo three-dimensional reconstruction method and device, electronic equipment and a storage medium, and relates to the technical field of embryo quality evaluation. Comprising the following steps: acquiring a multi-focal original plane image of a blastocyst to be reconstructed based on a time difference incubator; adopting a pre-trained image frame interpolation model to perform iterative frame interpolation processing on the multi-focal original plane image to generate a multi-focal extended plane image corresponding to each iteration; according to the multi-focus original plane image and the multi-focus extended plane image corresponding to each round of iteration, adopting an auxiliary circle marking algorithm to mark a blastocyst main body area in each focus original plane image and each focus extended plane image; respectively extracting mask images of a plurality of inner cell clusters and mask images of a plurality of trophoblast cells on the basis of marking results; and performing three-dimensional reconstruction according to the extracted mask images of the internal cell clusters and the mask images of the trophoblast cells to obtain curved surface models of the internal cell clusters and curved surface models of the trophoblast cells. The method improves the reconstruction precision.

The invention provides biopsy liquid and application thereof in pig blastocyst laser cutting, and belongs to the technical field of animal biology. The invention provides a biopsy solution and a preparation method thereof, the biopsy solution can be applied to pig blastocyst biopsy by laser cutting, the problem that at present, no adaptive biopsy solution exists for laser cutting of pig blastocyst is solved, and the invention further provides a method for laser cutting of pig blastocyst by using the biopsy solution. The problems of embryo cell injury, embryo development potential damage and the like possibly caused by laser cutting of the pig blastocyst in the prior art are solved. The biopsy liquid and the use method thereof can be efficiently and accurately applied to acquisition of pig blastocyst trophoblast cells, and are low in cost and good in repeatability.

Provided is a mammalian-derived cell model that can be used in anti-aging drugs. The mammalian-derived extraembryonic trophoblast cell model is an early mammalian-derived extraembryonic stage. A change model and an aging evaluation index system for differentiating the cells into the extraembryonic trophoblast cells / organoids can be used for various anti-aging researches and target detection, anti-aging drug detection, screening and identification, research on innovative drugs / natural products, and development of chemicals and / or health foods and additives. Screening and identifying many resistances is helpful. Candidate targets of aging may be further mechanically and functionally validated for future therapeutic use. The screening of anti-aging drugs or natural products will produce huge economic and social benefits.

The invention belongs to the technical field of biological medicine and molecular biology, and particularly relates to application of circ0005704 and related biological elements thereof in diagnosis and treatment of recurrent spontaneous abortion with unknown reasons. Researches find that differential expression and autophagy level of ULK1 in trophoblast cells of URSA patients are increased. High-throughput transcriptome sequencing is combined with bioinformatics analysis, and it is found that miR-26a-5p is down-regulated in URSA patients, is responsible for up-regulation of ULK1 and promotes autophagy of trophoblast cells, so that occurrence of URSA is increased. Through a high-throughput transcriptional set screening strategy and dual luciferase reporter gene analysis, it is found that circ0005704 enhances the expression of ULK1 through miR-26a-5p. In a word, the research of the invention shows that the circ0005704 / miR-26a-5p / ULK1 signal axis participates in the pathogenesis of URSA by adjusting the migration of the trophoblast cells, and a new target and scientific evidence are provided for the clinical treatment of URSA.

The invention belongs to the field of immunology, and particularly relates to amplification of NK cells, in particular to a method for amplifying NK cells by using NK92 cells as trophoblast cells, which specifically uses NK92 cells secreting IL-21, IL-15 and IL-18 as trophoblast cells and PBMC as seed cells to directionally amplify and activate natural killer cells, and comprises the following steps: transfecting an NK92 cell strain through lentivirus; the NK92 cell strain capable of stably expressing IL-21, IL-15 and IL-18 is obtained; then obtaining monoclonal cells of NK92 cells through flow sorting and limit dilution methods, finally culturing and amplifying, preparing NK92 trophoblasts through irradiation, and mixing the NK92 trophoblasts with PBMC (peripheral blood mononuclear cells) for culturing so as to prepare the NK cells. According to the method, the NK cells can be efficiently amplified and activated, and the activated NK cells can improve the immunity of the organism, so that tumor and virus infection is resisted. The invention has wide application in medicine.

The invention discloses a construction method of a trophoblast organ-like model, the trophoblast organ-like model and application thereof, and relates to the technical field of cell model.The construction method comprises the steps that placenta villus tissue is aseptically separated, gradient separation and purification are conducted to obtain extraembryonic early cells, the extraembryonic early cells are mixed with matrigel, and the trophoblast organ-like model is obtained; and carrying out 3D culture by using a culture medium containing early-stage and later-stage differentiation compositions to obtain the organ-like model with various trophoblast cell lineages. Based on the model, placenta toxicity research is carried out on pregnancy drugs, and the research specifically comprises the degree of 3D organoid form destruction after drug treatment, organoid activity and the expression condition of trophoblast marker genes. Compared with a traditional two-dimensional cell model, the placental toxicity detection method has the advantages that the placental toxicity of the medicine is detected by taking the trophoblast-like organ as the model, the accuracy is higher, the predictability is higher, and the influence of the medicine on human placental tissues is accurately reflected.

The invention discloses application of vitamin D3 in preparation of a product for improving trophoblast proliferation inhibition. The research proves that the vitamin D3 can obviously improve the proliferation inhibition of palmitic acid on human trophoblast cells; the vitamin D3 can reduce palmitic acid induced cell active oxygen level up-regulation; vitamin D3 can up-regulate expression of Nrf-2, HO-1 mRNA and protein in cells induced by palmitic acid. Vitamin D3 can improve palmitic acid induced human trophoblast cell proliferation inhibition by activating an Nrf2 / HO-1 pathway. The invention provides a new product for improving the proliferation inhibition of palmitic acid on human trophoblast cells, and has a good clinical application prospect.

The invention provides a trophoblast cell characteristic analysis method and electronic equipment, and relates to the technical field of embryo evaluation. The method comprises the following steps: carrying out space quadrant division on a target blastocyst three-dimensional model obtained by three-dimensional reconstruction to obtain a plurality of sub-quadrants; determining polar trophoblast cell information and wall trophoblast cell information according to the internal cell mass information in each sub-quadrant; determining multiple fusion feature data according to the trophoblast cell information, the polar trophoblast cell information and the wall trophoblast cell information in each sub-quadrant; and according to the fusion feature data and the basic feature data, carrying out embryo development result analysis to obtain estimated development result information of the target blastocyst. According to the scheme, the spatial structure features of the trophoblast cells are fully mined to obtain the fusion features, and the embryonic development result can be analyzed more deeply based on the fusion features, so that the accuracy of embryonic development result evaluation is improved, and clinical selection of embryos with higher development potential is assisted.

The invention discloses a trophoblast-free one-step culture method and application of tumor infiltrating lymphocytes, and particularly discloses a one-step amplification method of tumor infiltrating lymphocytes, which comprises the following steps: treating a tumor sample obtained from a subject into tumor segments; the tumor segments are cultured in a culture medium containing a CD3 agonist, a 4-1BB agonist and IL-2, so that an amplified tumor infiltration lymphocyte population is obtained, and the culture medium does not contain trophoblastic cells. Compared with the prior art, the method has the advantages that the required period for amplifying the tumor infiltrating lymphocytes is shorter, the cost is lower, the cell effect function is stronger, and the clinical application range of the tumor infiltrating lymphocyte therapy is widened.

The invention provides a biological preparation for regulating placenta dysplasia, and belongs to the technical field of gynaecology and obstetrics. Core effective components of the biological preparation are lutjanus mixed polypeptide with the molecular weight ranging from 2 kDa to 10 kDa, and the lutjanus mixed polypeptide is prepared by combining lutjanus fish bones, fish scales and fish flesh according to the mass ratio of 5: 3: 2 through the steps of desalting treatment, ultrasonic-assisted degreasing, double-enzyme synergistic enzymolysis, double ultrafiltration purification and the like. Experimental results show that the lutjanus mixed polypeptide has a remarkable protection effect on a human trophoblast cell line HTR-8 / SVneo in a high-glucose environment, and can effectively promote cell proliferation and migration and reduce high-glucose-induced cell damage. Therefore, the biological preparation provides a novel effective intervention means for placenta dysplasia caused by gestational diabetes mellitus, and has a good application prospect.

The present application belongs to the field of cell therapy and immunotherapy, and provides a genetically engineered human trophoblast, a preparation method and application thereof. The human trophoblast takes K562 cells as starting cells, and stably expresses membrane-bound interleukin 21, CD137 ligand and Delta-like ligand 1 after genetic engineering. The constructed K562 three-factor trophoblast can significantly improve the expansion efficiency, activation state and functional stability of NK cells and γδT cells. The synergistic mechanism includes enhancing the proliferation, cytotoxicity and stemness maintenance of NK cells and γδT cells through STAT3, NF-κB and Notch signaling pathways, respectively. The human trophoblast has the advantages of good expression stability, significant functional enhancement, and high activity after freezing and recovery.

An engineered trophoblast that expresses a cell surface molecule that specifically recognizes NKp46, the cell surface molecule comprising at least an antibody targeting NKp46 and a transmembrane domain. The invention discloses a cell culture medium and a method for amplifying NK cells. Compared with unengineered trophoblasts, the engineered trophoblasts are used for activating NK cells, and the amplification capacity of the NK cells and the killing activity of tumor cells can be enhanced.

The invention belongs to the technical field of biology, and particularly relates to application of baicalin to preparation of a medicine for treating preeclampsia by inhibiting trophoblast ferroptosis induced by NETs. The baicalin inhibits downstream signal transduction by specifically interfering a key activation step of PKC [beta] 2 and a membrane translocation process coupled with a second signal, and finally plays a role in inhibiting ferroptosis of trophoblasts. The discovery provides a brand new perspective for understanding the molecular action mechanism of the medicine. According to the application, baicalin influences ferroptosis induced by NETs in trophoblast cells by inhibiting membrane translocation and activation of PKC beta2, so that the biological function of baicalin is exerted. The discovery of the mechanism further deepens the important significance of people on NETs and baicalin in disease induction and treatment, and a thought is provided for research and development of a new treatment scheme for preeclampsia.

The invention belongs to the technical field of bioengineering, and particularly relates to application of RPS26 recombinant protein in promoting early embryonic development of animals. Experiments show that after the RPS26 recombinant protein is added in the early-stage in-vitro embryo culture process of the lamb goats, the blastocyst rate of the lamb goats can be increased to 40.0%; meanwhile, the cell proportion of cell mass in the blastocyst can be remarkably increased, the cell proportion of trophoderm is reduced, and the total number of blastocyst cells is not influenced, so that the blastocyst development rate can be remarkably increased and the blastocyst quality can be improved by utilizing the RPS26 recombinant protein.

The invention provides an application of DCTPP1 in detection of an auditing abortion kit, relates to the technical field of target spot diagnosis, and particularly provides an application of DCTPP1 as a target spot in preparation of an auditing abortion detection product, and an experimental result shows that DCTPP1 is one of key genes for auditing abortion. Specifically, after the DCTPP1 is knocked down from a human embryo trophoblast cell line HTR8 / SVneo, compared with a control group, the reduction of the DCTPP1 can cause more cell oxidation stress increase, and cell viability is reduced 24 hours later, and finally cell apoptosis occurs; in the mechanism, the DCTPP1 interacts with the RNA binding protein AUF1, and the reduction of the DCTPP1 causes the increase of oxidative stress of human embryo trophoblastic cells through the AUF1, so that apoptosis is caused. Experimental results in the application show that DCTPP1 is one of key genes for auditing abortion, and cell apoptosis can be finally caused by destroying the function of DCTPP1 in embryonic trophoblast cells through technologies such as transfection of specific siRNA and the like. In addition, the DCTPP1 knocked-down embryo trophoblast cells can be subjected to apoptosis, and the DCTPP1 knocked-down embryo trophoblast cells are one of useful genes for detecting and auditing abortion.

The present disclosure relates to antigen-binding molecules that bind to human and feline Human Trophoblast Cell Surface Antigen 2 (TROP2). The specific embodiments relate to antibody-drug conjugates (ADC) comprising saporin, mertansine and MMAE and chimeric antigen receptors (CAR) comprising said antigen-binding molecules for use in treatment of cancer.

The application relates to the field of biotechnology, in particular to application of CADM1 in preparation of a drug for gestational diabetes mellitus. The application comprises: using a detection reagent for detecting the expression level of CADM1 in preparation of a detection drug for gestational diabetes mellitus. The application is based on the previous research of the applicant that the expression level of CADM1 in the placental trophoblast of a gestational diabetes mellitus patient is reduced, and it is found in an in-vitro experiment that CADM1 can promote the proliferation, migration and invasion of the placental trophoblast cells, and by constructing an mRNA expression amount of CADM1 and a receiver operating characteristic curve of a subject of a GDM patient, it is indicated that the mRNA expression amount of CADM1 is related to the GDM patient, which indicates that CADM1 is related to the placental trophoblast of the gestational diabetes mellitus patient, and it is indicated that the detection of the gestational diabetes mellitus can be realized by detecting the detection reagent for detecting the expression level of CADM1.

The invention provides a trophoblast cell analysis method, device and equipment and a storage medium, and relates to the technical field of embryo quality evaluation. Comprising the steps of collecting a multi-focal plane image of a target blastocyst; performing image fusion on the multi-focal plane image based on a pre-trained image fusion model to generate a first fusion image and a second fusion image corresponding to the target blastocyst; performing main body region segmentation on the first fusion image and the second fusion image to obtain a first target image and a second target image; segmenting the trophoblast cells in the first target image and the second target image by adopting a pre-trained image segmentation model to obtain a first mask image and a second mask image; according to the first mask image and the second mask image, multiple feature parameters of the trophoblast cells are extracted, and based on the feature parameters, correlation between the feature parameters and the development result of the embryo is obtained through analysis. According to the method, a comprehensive interpretable feature analysis method is realized, and the analysis accuracy is improved.

Antibodies and antigen-binding fragments thereof specific to Trophoblast cell surface antigen 2 (Trop2). Also provided are anti-Trop2 antibody-drug conjugates (ADCs), Trop2-specific chimeric antigen receptor, and multispecific antigen binding proteins, such as bispecific T cell engagers (BiTEs), which bind to Trop2 and other targets. Further provided are compositions comprising the anti-Trop2 antibodies and antigen-binding fragments thereof, a method of diagnosis and cancer treatment employing the anti-Trop2 antibodies and antigen-binding fragments thereof.

The invention belongs to the technical field of biological medicine, and particularly relates to application of beta cell protein in preparation of medicine for preventing and treating abortion. Research finds and verifies for the first time that application of beta cell protein (BTC) promotes invasion and tubulation processes of extravillus trophoblast cell line-HTR8 cells, BTC has an obvious rescue effect on embryo absorption in abortion model mice in early and middle stages of pregnancy, and BTC is injected into the abortion model mice in an intraperitoneal cavity in early and middle stages of pregnancy to increase placenta weight, so that the abortion model mice in early and middle stages of pregnancy have an obvious rescue effect on embryo absorption. Therefore, the BTC has a certain treatment effect on the abortion, so that the BTC has a good practical application value.

The application provides a blastocyst three-dimensional fitting reconstruction method and device, electronic equipment and storage medium. The method comprises the following steps: acquiring a plurality of focal plane images of a blastocyst; identifying a trophoblast cell mask and an inner cell mass mask in the blastocyst based on the plurality of focal plane images; constructing a trophoblast cell grid model and an inner cell mass grid model according to the trophoblast cell mask and the inner cell mass mask; constructing a depth map of the blastocyst according to the plurality of focal plane images; performing image fusion on the plurality of focal plane images to obtain a texture image; performing plane mapping according to the texture image and the depth map to generate an internal three-dimensional point cloud model of the blastocyst; constructing a three-dimensional geometric model of the blastocyst according to the trophoblast cell grid model, the inner cell mass grid model and the internal three-dimensional point cloud model; and mapping the texture image to the surface of the three-dimensional geometric model to generate a blastocyst three-dimensional model. The reconstruction based on the method does not affect the development of the embryo, and is low in cost and safe and reliable.

The separation and detection method comprises the following steps: rinsing placenta or villus tissues in a PBS (Phosphate Buffer Solution) added with a 1% penicillin-streptomycin double antibody solution, digesting the placenta or villus tissues into single cells by using a 1640 culture medium containing I-type and IV-type collagenase after the placenta or villus tissues are rinsed sufficiently to obtain a cell suspension, and culturing the cell suspension for later use; diluting and neutralizing collagenase digestive juice by using a 1640 culture medium containing high-temperature heat-inactivated fetal calf serum according to 5 times of volume; repeatedly discarding the precipitate, recovering the supernatant, and centrifuging to obtain a coarse migration body; and establishing a density gradient by using Optiprep as a density medium, washing the precipitate with PBS, and centrifuging to obtain the trophoblast cell-derived transporter. The invention provides a group of placenta villus trophoblast cell source migration body specific expression membrane proteins for the first time, and provides a method for specifically capturing the placenta trophoblast source migration body on the basis of the placenta villus trophoblast cell source migration body specific expression membrane proteins, so that a new direction for researching a trophoblast dysfunction related disease mechanism can be enriched.

The invention provides an NK cell in-vitro efficient culture system and application, the culture system comprises a K562 engineering cell, and six proteins of IL21, IL12, IL15, CD40L, CD155 and CD112 are expressed on a cell membrane of the K562 engineering cell in a transmembrane mode; the NK cell culture composition comprises the following components: a liquid serum-free cell culture medium, 1-5% (v / v) of autologous plasma, 100-250 [mu] g / ml of an astragalus extract and 200-500 [mu] g / ml of an angelica sinensis extract, and the dosage ratio of the astragalus extract to the angelica sinensis extract is 1: 2. The NK cell with high amplification multiple, high purity and high lethality can be obtained by a culture system established by taking the K562 engineering cell constructed by the invention as a trophoblast cell, taking an astragalus extract and an angelica sinensis extract as culture additives and taking PBMC as an original culture material, and the culture system has important significance for treating cancers by cell drugs.

The invention discloses a culture medium for goat trophoblast organoid, a culture method and application, and belongs to the field of goat trophoblast organoid culture. The method comprises the following steps: separating trophoblast tissues from a healthy goat placenta which is pregnant for 80-90 days, digesting the trophoblast tissues through type II collagenase to obtain primary cells, compounding the primary cells with matrigel, and performing three-dimensional culture in an improved trophoblast organ culture medium with specific components to form mature organs. The organ is of a typical three-dimensional structure, and KRT23 and SOX2 markers are stably expressed. The organoid culture system based on the goat primary trophoblast cells is established for the first time, gene modification is not needed, the in-vivo physiological status is highly simulated, and the culture system is standardized and good in repeatability. The constructed organoid can be used as an ideal in-vitro model for researching placenta development, evaluating drugs and toxins and modeling infectious diseases in the gestation period, and is of great significance in improving the reproductive research level of goats and the production benefits of animal husbandry.

The invention discloses an NK (Natural Killer) cell in-vitro amplification culture method which comprises the following steps: on the 0th day, inoculating mononuclear cells into a culture bottle containing an NK cell culture solution, autologous plasma and an exosome of HEK293 cells, and standing and culturing in an incubator for 14 days; during the culture period, supplementing the NK cell culture solution and the autologous plasma once every two days, and controlling the cell density to be 1.0 * 10 < 6 > cells / mL; wherein the exosome of the HEK293 cell also needs to be supplemented on the seventh day; and after 14 days of culture, collecting centrifugal cells to obtain the NK cells. According to the culture method, the exosome of the HEK293 is used for replacing trophoblast cells added in the process of culturing the NK cells by a feeder method, so that the safety risk possibly caused by directly using the trophoblast cells is reduced; meanwhile, the HEK293 exosome is added and is combined with the NK cell surface receptor, so that synergistic stimulation is provided, the NK cells are activated and amplified at a high speed, and the multiplication capacity and purity of the NK cells are greatly improved.

The invention discloses an establishment method and application of a placenta trophoblast organoid model with correct physiological polarity, belongs to the technical field of organoid and stem cell model establishment, and aims to solve the problem of structural polarity error in trophoblast cells at present. The establishment method comprises the following specific steps: step 1, inducing nave embryonic stem cells to differentiate hTSC hollow spheres; step 2, transferring the hTSC hollow sphere into matrigel for culture to obtain a placenta trophoblast organ; and 3, transferring the placenta trophoblast-like organs into the 2D maintenance system to maintain the growth of the placenta trophoblast-like organs. According to the invention, the internal cavity of the placenta villus trophoblast and the physiological polar structure in which STB is outside and CTB is inside in trophoblast cells can be simulated, and the placenta trophoblast organ with three cell types of CTB, STB and EVT can be constructed at the same time.

The invention discloses an immortalized goat placenta trophoblast cell line and a construction method thereof, and relates to the technical field of cell engineering. The construction method comprises the following steps: carrying out isolated culture by using a tissue block culture method to obtain primary goat placenta trophoblast cells; eukaryotic expression plasmids pCI-neo-hTERT are introduced into the primary goat placenta trophoblast cells, positive clones are obtained through screening, subculture is carried out to 80 generations, and the immortalized goat placenta trophoblast cell line is obtained. The genetic structure of the immortalized goat placenta trophoblast cell line constructed by using the construction method can be kept stable in an infinite passage process, and when the immortalized goat placenta trophoblast cell line is transferred to 80 generations, the number of chromosomes is still 60, and the chromosomes are not lost or doubled. Transwell and RT-PCR detection prove that the immortalized goat placenta trophoblast cell line has an invasion characteristic, and TERT protein is stably expressed in the immortalized goat placenta trophoblast cell line.

The present invention relates to a method for activating and amplifying natural killer cells and uses thereof. Specifically, the present invention provides a method for activating and culturing NK cells, using a combination of recombinant proteins MICA and MICB as activation reagents to activate and culture NK cells in a coated state. The method of the present invention can achieve efficient amplification of NK cells without the addition of serum and trophoblast cells, and the amplification multiple can be as high as 800 times. The culture process of the present invention is simple, avoids high-risk materials, has high safety, and is particularly suitable for cells from sources such as iPSC-NK. The NK cells finally obtained have high purity and strong killing activity, and can be used to produce high-quality NK cell products.

The application discloses a simple and low-cost method for culturing canine NK cells in vitro and belongs to the technical field of cell culture. The method comprises the following steps: separating NK cells from peripheral blood of a dog, culturing the NK cells with a canine NK cell activation culture medium for 24-48 hours, then continuing to culture the NK cells with a canine NK cell expansion culture medium, adding the canine NK cell expansion culture medium after 48 hours, expanding the cells in a bottle when the cells are obviously expanded, otherwise maintaining the cells in the original bottle after centrifugation, adding liquid and expanding the cells in a bottle or culturing the cells in the original bottle every other day, and harvesting the canine NK cells after 12-18 days. The method provided by the application does not need to activate the canine NK cells by using engineered trophoblast cells and multiple cell factors, and does not need to activate the cells by using an antibody, so that the method is simple and low in cost, and the cultured canine NK cells normally express NKp46, do not express CD3 and CD5, and have normal inhibitory and killing tumor abilities.

The present invention belongs to the field of medical technology and specifically relates to the use of a mulberry leaf aqueous extract in improving adverse pregnancy outcomes caused by PM2.5. The mulberry leaf aqueous extract can effectively improve adverse pregnancy outcomes caused by PM2.5; the mulberry leaf aqueous extract prevents and treats adverse pregnancy outcomes caused by PM2.5 by reducing the biological damage to trophoblast cells caused by PM2.5.