45 results about "In vitro incubation" patented technology

The present invention relates to a medium composition for in vitro fertilization and / or in vitro culture comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, and a method using same. The compound or composition suppresses reactive oxygen species and lipid peroxidation in ovarian granulosa cells to preventing cytotoxicity and mitochondrial dysfunction caused by apoptosis and ferroptosis, leading to an improvement in the quality of aged oocytes, and to increase the developmental rate and blastocyst formation rate of pre-implantation embryos, thereby improving the efficiency of in vitro fertilization. In addition, treatment with the medium composition during the vitrification process of in vitro–fertilized embryos improves re-expansion and survival rates, thereby reducing structural and metabolic damage. Accordingly, the present invention can be advantageously applied to an assisted reproductive technology for treating infertility and subfertility, and in vitro fertilization and / or in vitro culture for improving the propagation efficiency of livestock.

This invention discloses an in vitro maturation culture medium for ferret oocytes and its preparation method, belonging to the field of animal embryo engineering technology. It comprises the following components: TCM199 basal culture medium, 50 mL fetal bovine serum, 25 IU follicle-stimulating hormone, 25 IU luteinizing hormone, 0.5 mg 17β-estradiol, 0.005 mg epidermal growth factor, 12.1 mg sodium pyruvate, 0.2 mM cysteine, 0.1 mM cysteine, 0.030 g penicillin, 0.025 g streptomycin sulfate, as well as an antioxidant complex, a combination of growth factors, and extracellular matrix components. This invention provides a comprehensive and suitable nutritional environment for ferret oocytes by precisely formulating a composite culture medium containing multiple key components. Specifically, the hormone combination at specific concentrations accurately simulates the in vivo physiological environment, effectively regulating the oocyte maturation process; multiple growth factors work synergistically to promote oocyte growth, development, and differentiation; and the addition of the antioxidant complex effectively eliminates free radicals generated during in vitro culture, reducing oxidative stress damage to oocytes.

The application provides a kind of liposome that can enhance organ targeting, preparation method and application, belongs to the field of biological medicine and nanotechnology.The application first proposes the strategy of actively regulating LNP organ targeting by pre-assembling specific protein crown in vitro, which breaks through the technical limitations of traditional passive formation of natural protein crown.Through selecting main lipids (CDO, CDA, EDM) with different head chemical structures, precise control of protein crown composition is realized.Furthermore, the method of the application is simple in process and mild in condition, only needs standard in vitro incubation steps, and has great clinical transformation potential.The pre-assembled protein crown method provided by the application significantly enhances the organ targeting efficiency of LNP: after pre-incubation of LNP1 with APOA1, the enrichment degree of spleen is increased by 2.8 times;after pre-incubation of LNP2 with VTN, the enrichment degree of lung is increased by 52%;after pre-incubation of LNP3 with APOE, the enrichment degree of liver is increased by 1.2 times.

The invention relates to a composition for making a tea beverage, the composition comprising tea which may be ground tea in combination with plant cell culture grown in vitro in a large scale. The plant cell culture grown in vitro in a large scale may be derived from any plant, such as a fruit including but not limited to grape berry, pomegranate, olive, and the like. The composition may be inserted into a pod and be used in a machine or inserted into a tea bag or a container.

This invention discloses a method and system for embryo quality detection, relating to the field of embryo detection technology, including the following steps: S1: fertilized embryos are continuously cultured in an in vitro culture system containing specific metabolic marker detection components, and continuous dynamic imaging of the embryos is performed to obtain a complete dynamic developmental image sequence; S2: the dynamic developmental image sequence is segmented, and temporal developmental features are captured, identifying key time points such as pronucleus appearance and disappearance, cleavage, and blastocyst formation, and generating a segmentation mask corresponding to each frame of the dynamic developmental image sequence; this invention achieves full-process time-difference imaging to capture the temporal features of embryonic development, without trauma or interference, avoiding subjective bias in static observation; by segmenting the dynamic developmental image sequence and capturing temporal developmental features, the segmentation accuracy is high and the anti-interference ability is strong.

This invention belongs to the field of tumor biology and drug screening technology, specifically disclosing a human EGFR mutation-driven mouse primary lung cancer cell line, its construction method, and its applications. The cell line, ZST-1, is a human EGFR (L858R / T790M) mutation-driven lung cancer cell line derived from mouse primary lung cancer. It is stable, capable of subcutaneous tumor formation in C57BL / 6 mice, and simultaneously expresses Luciferase and tdTomato reporter genes. Its construction method includes obtaining transgenic mice, virus-induced tumor formation, continuous in vivo passage in nude mice, and in vitro culture and screening steps. This cell line can be applied to in vitro screening and efficacy evaluation of human EGFR mutation-targeting drugs, research on EGFR-TKI resistance mechanisms, tumor bioluminescence imaging and fluorescence tracing, and in vivo tumorigenesis and efficacy experiments in an immune-intact C57BL / 6 background.

The application discloses a Toxoplasma SRS29C and ROP16 double-gene deletion strain and a construction method and application thereof, relates to the technical field of parasite genetic engineering and molecular biology. The Toxoplasma SRS29C and ROP16 double-gene deletion strain is obtained by knocking out SRS29C and ROP16 genes of the ME49 strain through a CRISPR / Cas9 system. The double-gene deletion strain constructed by the application has drug sensitivity change (sensitivity to ethidium bromide), growth defect phenotype, and only carries a single screening marker, and under in-vitro culture conditions, the double-gene deletion strain presents a significant slow growth trend, and the number of worm-containing bubbles is obviously less than that of the wild type and single-gene deletion strain. The double-gene deletion strain substantially influences the proliferation of the worm body, and provides key data support for exploring the molecular basis of the double-gene deletion leading to the increase of the drug sensitivity of the worm body and for analyzing the regulation mechanism of the drug tolerance related path of the Toxoplasma.

The application relates to the technical field of biology, and discloses a method for inducing a cell line of immortalized musk deer scent glands to synthesize dihydrotestosterone, wherein the cell line is obtained by transfecting a primary musk deer scent gland epithelial cell with a lentivirus carrying an SV40 Large T antigen and performing resistance screening; the cell line is in an epithelial cell form and stably expresses keratin 18, cholesterol side chain cleavage enzyme, 17 alpha-hydroxylase and 3 beta-hydroxysteroid dehydrogenase. Through lentivirus-mediated SV40 Large T antigen transfection, the primary musk deer scent gland epithelial cell breaks through the limit of replicative senescence and obtains unlimited proliferation capacity, effectively overcomes the defects that the primary cell is prone to fibrosis and rapidly loses metabolic function during in-vitro culture, provides a standardized cell model with a clear genetic background and persistent metabolic activity for biological manufacturing of dihydrotestosterone, and relieves the dependence on endangered animal live resources.

This invention relates to the field of livestock and poultry breeding and food safety testing technology, and in particular to a method for assessing copper exposure hepatotoxicity based on the endoplasmic reticulum stress-endoplasmic reticulum autophagy axis and its application. This method obtains liver tissue samples or in vitro cultured hepatocyte samples from copper-exposed subjects, and jointly detects the expression levels of key markers of the endoplasmic reticulum stress pathway and characteristic markers of the endoplasmic reticulum autophagy pathway. Combined with two-way pharmacological validation, a grading assessment standard for copper exposure hepatotoxicity is established. This invention overcomes the shortcomings of traditional liver enzyme indicators, such as low sensitivity and strong lag, and has the advantages of early warning, high specificity, and reliable results. It can be widely applied to scenarios such as determining the copper safety threshold in livestock and poultry feed, early screening for copper exposure hepatotoxicity, assessment of copper pollution ecological risks, and screening of copper toxicity protectants, providing technical support for ensuring animal health and the safety of animal-derived food.

The utility model discloses a fertilized egg in vitro incubator, including incubator, the incubator is slid and is provided with a plurality of groups of drawers, a plurality of groups drawer all are fixedly connected with the placement ring, the bottom fixed mounting of placement ring has the buffer pad, the drawer is fixedly connected with the limiting block, the both sides wall of incubator all are fixedly connected with the connecting plate, the one side fixed mounting of connecting plate has the sliding block, the outside slide of sliding block is provided with slide rail, the slide rail fixed connection is in the side of drawer, the one end fixed mounting of connecting plate has the damper, in the utility model, through the speed of reducing drawer to the inside movement of incubator, reduce the impact force that the drawer is contacted when closing drawer with incubator box body produces the influence to drawer, thereby reduce the vibration of the petri dish of placing on the drawer, avoid the culture solution in petri dish splash or the fertilized egg shift, guarantee the culture effect of fertilized egg, improve the practicality of incubator.

The present application relates to the technical field of reproductive biology, and specifically discloses a method for culturing mammalian ovary tissue in vitro based on dynamic regulation of gas-liquid interface. In view of the problem that oxygen supply and nutrient supply cannot be optimized and dynamically regulated in the existing in vitro culture of ovary, the method comprises the following steps: preparing an agar block with culture holes; controlling the liquid level of the culture solution to be flush with the outer edge of the agar block and not higher than the upper surface, so that the culture holes are initially liquid-free; placing the mammalian ovary in the holes and directly exposing it to the gas phase environment in the initial stage; during the culture process, the culture solution gradually enters the holes through capillary action, so that the ovary automatically changes to a semi-submerged state, and dynamic changes of the gas-liquid interface are realized. The present application does not require feeder cells or complex scaffolds, and is simple to operate. Through the synergistic effect of initial gas phase exposure and subsequent semi-submerged state, the internal hypoxia of the ovary is significantly improved, cell apoptosis is reduced, and follicle assembly is promoted, so the present application can be used for follicle development mechanism research and reproductive toxicity evaluation.

The present invention relates to a method for in vitro culture of frozen-thaw fertilized eggs using an in vitro culture medium containing 1α,25-Dihydroxyvitamin D3.

The embodiments of the present application provide a method for establishing in vitro CD8 + T cell exhaustion model and application thereof. The method comprises: using T cell receptor signal stimulator combined with interleukin-10 to continuously stimulate CD8 + T cells under in vitro culture conditions. The cells induced by the method not only highly express exhaustion key transcription factor TOX and various surface inhibitory receptors (such as PD1, TIM3, LAG3 and the like), but also significantly reduce the secretion ability of effector cytokines, highly reduce the real exhaustion performance in in vivo chronic infection or tumor microenvironment in vitro, and effectively overcome the defect of few CD8 + T cells in traditional in vivo animal model, and high-quality exhaustion cells can be obtained in vitro in large scale in only 6 to 8 days.

The application discloses a culture solution for improving the blastocyst rate and quality of bovine embryos, application and an in-vitro culture method, and relates to the technical field of animal embryo engineering. The application provides a culture solution for in-vitro culture of bovine embryos, which comprises a first culture solution and a second culture solution with different osmotic pressure concentrations. By controlling the osmotic pressure concentrations of the first culture solution and the second culture solution in a suitable range, and by using the first culture solution in the early stage of embryo development and the second culture solution in the late stage of embryo development, the in-vitro culture system can better simulate the environment in the oviduct in the early stage of embryo development and better simulate the environment in the uterus in the late stage of embryo development, so that the osmotic pressure stress of the embryo in the development process is reduced, and the blastocyst rate and the blastocyst quality of the bovine embryo are improved. Compared with a traditional culture system with constant osmotic pressure, the application significantly improves the blastocyst rate by sequential regulation of the osmotic pressure, the total cell number of the obtained blastocysts is more, and the apoptosis index is lower.

This invention relates to the NBLS (Niobium Bovine Kidney Cell Line), with accession number CCTCC NO: C2022197. This invention also provides its application in culturing BVDV virus. This invention successfully introduced the SV40-LT gene into primary Niobium Bovine Kidney Cells, activating their telomerase activity and extending the cell lifespan in vitro. Experiments showed that the NBLS Niobium Bovine Kidney Cell Line of this invention can be passaged for more than 50 generations without changing its growth and proliferation characteristics, thus obtaining an immortalized cell line. This F50 generation Niobium Bovine Kidney Cell Line NBLS is highly sensitive to BVDV virus and can efficiently proliferate the virus. The virus titer proliferated using this cell line shows increased sensitivity compared to primary Niobium Bovine Kidney Cells, reducing production costs. Therefore, this invention provides a new technical means for the large-scale production of BVDV vaccines.

This invention belongs to the field of biomedicine and reproductive health technology, specifically relating to the application of wheat germ extract in the preparation of products that improve the quality of aging oocytes. Through in vitro culture and in vivo gavage models, this invention demonstrates for the first time that wheat germ extract can significantly improve the in vitro maturation rate and in vivo superovulation maturation rate of oocytes from aged mice, significantly reduce intracellular reactive oxygen species levels, inhibit early apoptosis, and reduce oocyte mortality. The optimal in vitro concentration of the wheat germ extract is 0.1%–0.2%, and the optimal gavage dose is 5%. At these concentrations, the improving effect of wheat germ extract is significantly superior to that of spermidine. This invention provides a safe, natural, and long-term-accessible reproductive aging intervention strategy with good scientific value and promising prospects for industrial application.

This invention discloses a method for preparing a ternary dynamic glycopeptide hydrogel bio-ink, comprising the following steps: dissolving methacrylamide gelatin, aldehyde-modified hyaluronic acid, hydrazide-modified gelatin, and a photoinitiator together in a solvent and mixing them evenly to obtain the bio-ink. This bio-ink constructs a dynamic covalent cross-linking network through reversible acylhydrazone bonds formed between aldehyde-modified hyaluronic acid and hydrazide-modified gelatin, providing a remodelable three-dimensional microenvironment for cells. Simultaneously, the photocrosslinking properties of methacrylic anhydride-modified gelatin are introduced to rapidly solidify and maintain overall shape stability during the printing stage, providing a remodelable cellular microenvironment during subsequent in vitro culture. This establishes an organoid culture platform with both good printability and excellent cell compatibility, providing a reliable material basis for subsequent drug screening and functional studies.

Embodiments described herein relate generally to devices, apparatuses, and systems with embedded electrodes for growing, maintaining, and / or using 3D tissues in vitro. The devices, apparatuses, and systems described herein can provide scalable, automated tissue stimulation.

The present application relates to a method for improving the quality of platelets produced in vitro. Specifically, the present inventors provide a platelet culture medium for in vitro culture, which can significantly improve the yield and quality of platelets cultured in vitro by optimizing the ingredients and adding new ingredients. Specifically, the viability of platelets obtained by culturing megakaryocytes in the culture medium of the present application and the proportion of CD42b+ cells are significantly increased, and the number of platelets produced by a single megakaryocyte is also significantly increased. The present application improves the problems of imperfect platelet function and unstable quality in existing in vitro platelet production technology, provides a stable platelet source for clinical transfusion, and has a wide application prospect.

This invention relates to the field of biomedical engineering technology and discloses a method for the circular expansion of exosomes from nucleus pulposus cells and its application, comprising the following steps: S1: culturing HUMSCs and NPs separately, collecting the culture supernatant of the NPs and purifying and extracting hNP-exo; S2: diluting the hNP-exo with complete HUMSC culture medium and co-culturing it with in vitro cultured spare HUMSCs to induce the HUMSCs to differentiate into NP-like cells; S3: culturing the NP-like cells, collecting their supernatant and separating and purifying them to obtain NP-like exosomes. This method for the circular expansion of exosomes from nucleus pulposus cells and its application uses human umbilical cord mesenchymal stem cells (HUMSCs) as seed cells to replace traditional bone marrow mesenchymal stem cells (BMSCs). HUMSCs can be obtained from the umbilical cord, which is readily available and can be recycled with the consent of the mother and her family. The acquisition difficulty of HUMSCs is much lower than that of BMSCs, thus solving the defects of BMSCs, which require bone marrow aspiration, involve risks and pain, and have limited collection volume.

This invention relates to the field of platelet production technology, and in particular to a method for producing platelets based on biomimetic microcarriers. The method overcomes the key bottlenecks of low in vitro platelet yield, incomplete function, and difficulty in large-scale production in the prior art. By constructing a biomimetic bone marrow-vascular wall microenvironment, it systematically solves the technical problem of low yield of megakaryocytes in in vitro culture systems, and provides a brand-new solution for the safe, efficient, and economical in vitro preparation of platelets.

The present application provides a method for preparing human ovarian somatic cell-like cells. The present application establishes a method for preparing human ovarian somatic cell-like cells in vitro, which comprises stage-based culture based on human stem cells (including human pluripotent stem cells and human totipotent stem cells). The method of the present application can efficiently obtain functional human ovarian somatic cell-like cells, promote the differentiation ability of female germ stem cells (FGSC) cultured in vitro, and provide a new approach for assisted reproduction and treatment of infertility.

The present invention relates to an in vitro method for cultivating one or more antibody expressing cell(s). The method comprises cultivating one or more antibody expressing cell(s) obtained from peripheral blood in the presence of IL-2, IL-21 and a non-cell surface presented CD40-stimulating agent and in the absence of feeder cells. Moreover, herein provided are methods for producing antibodies comprising the step of cultivating one or more antibody expressing cell(s) according to the method of the invention, a novel CD40-stimulating agent and uses therefrom as well as a cell culture medium.

This application discloses an in vitro reconstructed vaginal mucosa model, its construction method, and its application. The construction method includes an improved culture medium: HVU medium is added to immortalized vaginal epithelial cell line VK2 / E6E7, which is then seeded into Transwell chambers and cultured. The HVU medium on the surface of the culture in the Transwell chambers is aspirated, and the Transwell chambers are raised to the air-liquid interface for air-liquid surface culture. The culture medium is then replaced with HVA medium to obtain the vaginal mucosa model. This method improves the stability of cells during in vitro culture and constructs an in vitro reconstructed vaginal mucosa model with a tissue structure highly similar to human vaginal mucosa (possessing a complete stratified squamous epithelial structure and normal intercellular connections), thereby enhancing the model's physiological functional simulation.

The application discloses a preparation method of cat induced pluripotent stem cells, which comprises the following steps: constructing piggyBac transposon plasmids carrying cat source OCT4, SOX2, KLF4 and c-MYC four factors in series, piggyBac transposon plasmids carrying cat source or pig source NANOG and LIN28 two factors in series and piggyBac transposon plasmids carrying monkey kidney virus SV40LT plasmids, and transfecting the plasmids into cat fetal fibroblasts; picking up cloned cells with embryonic stem cell morphologies, and carrying out subculture of cell clones meeting the characteristics of embryonic stem cells through identification and screening, so that the cat induced pluripotent stem cells are obtained. The preparation method of the cat induced pluripotent stem cells is safe, efficient and has good differentiation potential, lays a scientific foundation for researching in-vitro culture conditions of feline ESCs and establishment of feline iPSCs, and lays a scientific foundation for further cultivating cloned cats and gene modified cats and researching in related disease mechanisms, screening drugs and regeneration and repair.

Embodiments described herein relate generally to devices, apparatuses, and systems with embedded electrodes for growing, maintaining, and / or using 3D tissues in vitro. The devices, apparatuses, and systems described herein can provide scalable, automated tissue stimulation.

The application discloses a construction method of Aedes albopictus Aa01 cell line and susceptible cell model of Decapod iridovirus 1, and belongs to the technical field of virology and aquaculture disease prevention and control, wherein the Aedes albopictus Aa01 cell line is preserved in the China Center for Type Culture Collection of Wuhan University, and the preservation number is CCTCC NO: C202648. The method for constructing the DIV1 susceptible cell model based on the above cell line comprises the following steps: culturing the Aedes albopictus Aa01 cell line, and subculturing or using for virus inoculation when the density reaches 80%-90%; inoculating the DIV1 into the Aedes albopictus Aa01 cell line, removing the virus liquid after virus adsorption, and adding cell maintenance liquid for continuous culture; and observing the cytopathic effect CPE, collecting the cell culture when the CPE reaches 70%-80%, and obtaining the amplified DIV1 virus liquid. The DIV1 susceptible cell model constructed by the method can solve the problem that there is currently a lack of stable and continuously subcultured DIV1 in vitro culture.