15 results about "Murine embryo" patented technology

The invention belongs to the field of medicines, and relates to a compound derived from Sinkiang lithospermum, a preparation method thereof and application of the compound in prevention and treatment of cardiovascular diseases. Specifically, the invention discloses a monoterpene benzoquinone derivative with a novel structure in Sinkiang lithospermum, a preparation method of the monoterpene benzoquinone derivative and application of the compound in preparation of anti-myocardial injury drugs. An in-vitro anti-myocardial cell injury activity experiment proves that the compound has a remarkable protection effect on isoprenaline (ISO)-induced H9C2 cell (rat embryo myocardial cell) injury and hypoxia-reoxygenation-induced H9C2 myocardial cell injury. The result shows that the compound can be used for preparing the anti-cardiovascular disease medicine.

The invention belongs to the technical field of biomedicine, and relates to a method for constructing a mouse embryo-derived hypothalamic organ model and application, the invention proposes that a mouse embryo is used for constructing a hypothalamic organ for the first time, and the mouse embryo has a definite development time sequence and genetic operability; the naturally retained vascular endothelial cells and multilineage precursor cells provide unique advantages for constructing a three-dimensional organ containing multiple nucleuses such as a preoptic region, a paraventricular nucleus, an arcuate nucleus and a supraoptic nucleus, and the model not only breaks through the nucleus coverage limitation of the existing iPSCs organ, but also can be used for constructing the three-dimensional organ containing multiple nucleuses such as the preoptic region, the paraventricular nucleus, the arcuate nucleus, the supraoptic nucleus and the like by reconstructing a nerve-blood vessel unit. High-fidelity simulation of complex physiological functions of the hypothalamus is realized, and an irreplaceable technical platform is provided for mechanism analysis of neuroendocrine diseases and high-throughput screening of drugs.

This invention discloses a method for establishing and validating a hydrogen peroxide-induced mouse autism model, relating to the field of medical biotechnology. The method for establishing the hydrogen peroxide-induced mouse autism model includes the following steps: injecting the hydrogen peroxide solution into the peritoneal cavity of pregnant mice to induce hydrogen peroxide, resulting in offspring mice with hydrogen peroxide induction, i.e., the mouse autism model. This invention uses near-peritoneal injection of hydrogen peroxide during the gestational period of the mother mouse, inducing oxidative stress in both the pregnant mother mouse and the offspring embryo through hydrogen peroxide alone, without causing a significant immune response in the mother mouse. Therefore, an oxidative stress-specifically induced ASD mouse model is formed. The resulting ASD offspring mice exhibit significant ASD phenotypes but do not show obvious motor impairment.

This invention belongs to the field of gene editing and epigenetics technology, and discloses a method based on dCasRx-FTO for m 6 A. Editing system and its recombinant expression plasmid, cell model and its application; this invention first constructs an m based on dCasRx-FTO. 6 The A-editing system, namely the dCasRx-FTO fusion protein, utilizes catalytically inactivated CasRx (dCasRx) as a highly specific RNA guidance scaffold to fuse with m 6 A demethylase FTO enables the targeting of a specific region m 6 Targeted removal of A-modification. Building upon this, the present invention further obtains monoclonal mouse embryonic stem cells (mESCs) stably expressing the dCasRx-FTO fusion protein. Combined with an sgRNA library targeting a specific set of transcripts, high-throughput screening can be performed on embryonic stem cells containing the 2C-like pluripotency reporter gene MERVL::tdTomato, thereby identifying key m-modification inhibitors suppressing the pluripotency state of mESCs at the transcriptome level. 6 A modification site, thus enabling its use to reveal m 6 The key role of A-modified targets in the regulation of stem cell pluripotency.

The invention discloses application of ethyl caffeate in preparation of heart and nerve protection medicines, and belongs to the technical field of biological medicines. Models of rat embryo myocardial cell injury caused by oxygen-glucose deprivation and mouse microglial cell injury caused by lipopolysaccharide are respectively constructed, and related indexes such as inflammation, oxidative stress and cell apoptosis are evaluated; it is proved that ethyl caffeate has an obvious protection effect on ischemic and hypoxic myocardial cells and inflammation-damaged glial cells, and can be used for preparing heart and nerve protection drugs.

The invention provides a culture method for promoting conversion of mouse embryonic stem cells into totipotent stem cells. The cells can be kept in a brand new form and subcultured under the condition. According to the method adopted by the invention, an inhibitor WM-3825 of 2 mu M H4K16ac is added into a DMEM (Dulbecco Modified Eagle Medium) with a culture medium composition of 10% fetal calf serum according to a proportion of 1: 1000, the mouse embryonic stem cells can be normally cultured and passaged, the change of the totipotent gene is detected through real-time fluorescent quantitative PCR (Polymerase Chain Reaction), the expression of the totipotent related gene Dux is found to be obviously increased, and the expression of the mouse embryonic stem cells is obviously improved. Flow cytometry detection shows that the proportion of the totipotent stem cells is remarkably increased, and AP dyeing shows that the pluripotency of the mouse embryonic stem cells is reduced. The method starts from cell culture, and the method is possibly suitable for research on conversion from pluripotency to totipotency of embryonic stem cells of other mammals including human. By adding a small-molecule inhibitor WM-3825 of H4K16ac, a brand-new method for promoting conversion from the mouse pluripotent stem cells to the totipotent stem cells is established, and the 2-cell period of mouse embryonic development can be simulated in vitro through the culture system. The invention provides a new method for the generation of the totipotent stem cells, deepens the understanding of people on different states of the stem cells, and provides a new perspective for understanding an epigenetics mechanism between the conversion of the totipotent stem cells and the conversion of the pluripotent stem cells.

The invention discloses an induced differentiation and identification method for obtaining nerve cells in different stages from mouse embryonic stem cells, and relates to the technical field of biology. The induced differentiation and identification method comprises the following steps: inoculating mouse embryonic stem cells into a culture container, and carrying out pluripotent maintenance culture by adopting a pluripotent maintenance culture medium; after the pluripotency maintenance culture is completed, discarding the culture medium, washing, and adding a nerve induced differentiation culture medium for nerve induced differentiation culture; in the process of nerve induced differentiation culture, gene marker and protein marker detection is carried out to obtain nerve cells in different stages. According to the induced differentiation and identification method provided by the invention, efficient and controllable differentiation of the mouse embryonic stem cells to the neural pedigree can be realized through standardized sequential control and a multi-dimensional verification system. According to the invention, an efficient and reliable cell source and a technical platform are provided for the research of a neurodevelopment mechanism and the construction of a neurodegenerative disease model.

The method of generating multipotent stem cells is a method for producing and / or expanding multipotent stem cells by delivering at least one reprogramming protein into somatic cells. The at least one reprogramming protein includes a Master Regulator (MR) protein, which may be BAZ2B, ZBTB20, ZMAT1, CNOT8, KLF12, DMTF1, HBP1, or FLI1. The bromodomain protein BAZ2B, in particular, was identified by first generating bi-species heterokaryons by fusing Tcf7l1− / − murine embryonic stem cells (ESCs) with human B-cell lymphocytes. Reprogramming of the B-cell nuclei to a multipotent state was tracked by human mRNA transcript profiling at multiple timepoints. Interrogation of a human B-cell regulatory network with gene expression signatures collected from such reprogramming time series identified eight candidate Master Regulator proteins, which were validated in human cord blood-derived hematopoietic progenitor and lineage-committed cells.

The invention relates to the technical field of biological medicines, and discloses an application of phosphodiesterase PDE4D in preparation of a medicine for treating virus infectious diseases, and an application of a phosphodiesterase PDE4 inhibitor or a salt thereof in preparation of a medicine for treating virus infectious diseases for a long time. Experiments prove that the expression of PDE4D can significantly inhibit the replication of VSV, HSV, PR8 and other viruses; the cell death can be obviously inhibited; the inflammatory response caused by virus infection can be inhibited; the PDE4 inhibitor can induce the expression of PDE4D after long-term treatment; long-term treatment with the PDE4 inhibitor can significantly inhibit embryo loss of pregnant rats caused by virus infection; long-term treatment with the PDE4 inhibitor can reduce viral load of lung and placenta tissues of pregnant rats and pathological changes caused by viral infection.

This invention discloses a method for inducing mouse embryonic stem cells to transition to a pluripotent state using H2BC21 and its application, belonging to the fields of molecular biology and stem cell engineering technology. By constructing an interference vector targeting the H2BC21 gene, packaging it with lentivirus, and infecting mouse embryonic stem cells, the expression of the H2BC21 gene is knocked down, and embryonic stem cell lines with stable low expression of H2BC21 are obtained through screening. This invention is the first to discover that H2BC21 is a core negative regulator of mammalian embryonic stem cell pluripotency. After knocking down H2BC21, the expression of pluripotency core genes Nanog, Oct4, and Sox2 is downregulated, while the expression of pluripotency core genes Zscan4b, Usp17la, and Tmem92 is upregulated. The cells exhibit molecular characteristics similar to 2-cell phase cells, successfully achieving the transition from pluripotency to a pluripotent state.

The utility model relates to the technical field of mouse embryo culture, in particular to a mouse embryo incubator with a temperature control function, which comprises a rack and the like, the rack is a support carrier of the mouse embryo incubator and is a square cabinet, and a plurality of storage compartments are arranged in the rack at intervals in an array manner; the hatching box bodies are installed in the compartments of the rack, and openings for placing mouse culture dishes are formed in the front sides of the hatching box bodies; the fixing blocks are symmetrically and fixedly arranged on the two sides, close to the opening, of the front side of the incubation box body. The motor drives the lead screw to adjust the position of the heat insulation plate, the interior of the rack can be flexibly divided into two incubation compartments with different temperatures, independent temperature control over each compartment is achieved through the cooperation of the miniature air conditioners on the two sides, the most suitable temperature environment is provided according to different development stages of mouse embryos, and the incubation efficiency is improved. The success rate and the consistency of embryo culture are improved.

The invention belongs to the technical field of animal models, and particularly relates to a preparation method for constructing a rat embryo implantation failure model based on streptococcus agalactiae. The method comprises the following steps: culturing a streptococcus agalactiae strain with the number of BNCC336970 by using a solid culture medium containing sheep blood, adjusting the concentration of a bacterial solution to 1 * 10 < 8 > CFU / mL, and carrying out vaginal perfusion on an SPF-grade female SD rat every day for 21 days continuously. And after the perfusion is finished, the female mouse is mated with the male mouse at the early oestrus stage, the detected vaginal suppository is marked as the 0.5 th day of pregnancy, the mouse is killed at the 7.5 th day of pregnancy, and the model is evaluated by counting the embryo implantation quantity and detecting the expression level of endometrial tissue receptivity factors MMP9 and VEGF (Vascular Endothelial Growth Factor). A clinical pathogen pathogenic mechanism is simulated through a genital tract infection way for the first time, and the problem of physiological status deviation caused by traditional hormone intervention, immune agonists or genetic engineering methods is solved. The constructed model can stably induce the obvious reduction of the embryo implantation number and the reduction of endometrial receptivity, and provides a standardized tool for the pathological mechanism research of infectious embryo implantation failure and the screening of therapeutic drugs.

The utility model relates to the technical field of mouse embryo culture, in particular to a mouse embryo culture device which comprises a box body and a door plate, the box body is an assembly carrier of the culture device, an opening for placing a culture dish is formed in the front side face of the box body, and the door plate is hinged to the opening in the front side of the box body. A handle is mounted on the outer wall of the front side of the door plate; the motor is fixedly mounted on an inner bottom plate of the box body, and a convex rod is fixedly arranged on an output shaft of the motor; a hollow sleeve rod is fixedly connected to the bottom face of the rotating table, a sliding groove is formed in the side wall of the sleeve rod, and the rotating table is arranged on an output shaft of the motor through the sleeve rod. According to the utility model, the rotating table is driven by the motor to rotate slowly, and the culture dishes on the rotating table are intermittently shaken by combining the design of the concave-convex sliding rails, so that mechanical stimulation and flowing conditions similar to an in-vivo environment are created, and the development quality of embryos is favorably improved.

The invention discloses a method for constructing a rat embryo implantation failure model based on staphylococcus aureus, and belongs to the technical field of animal models. According to the method, ATCC23235 (purchased from ATCC with the strain number of ATCC 23235) staphylococcus aureus bacterial liquid (1 * 10 < 8 > CFU / mL, 200 [mu] L / mouse, and continuous 28 days) is perfused in vagina to induce SD rat embryo implantation number reduction and endometrial receptivity related factor expression reduction, and the pathological state of embryo implantation failure is successfully simulated. The model disclosed by the invention avoids limitation of hormone or gene intervention, is closer to a clinical infection mechanism, and is suitable for etiological research and therapeutic drug screening of embryo implantation failure.

This invention relates to an energy drug for treating mitochondrial damage under hypoxic conditions. The energy drug is the application of racemic scopolamine (Rac) in remodeling mitochondrial function. Hypoxic conditions include physiologically or pathologically induced hypoxia such as high altitude or vascular occlusion. This invention uses racemic scopolamine to treat hypoxic rat embryonic cardiomyocyte H9C2 and BALB / c mouse models, demonstrating that the compound has the characteristic of remodeling mitochondrial function under hypoxic conditions. It is a highly effective energy drug with promising clinical application prospects.