38 results about "Murine embryo" patented technology

The invention belongs to the field of artificial intelligence, and particularly relates to a mouse embryo organ recognition and scoring method and a system. The method comprises the following steps: collecting original images of mouse embryos in different periods, and carrying out the manual marking; inputting the mouse embryo original image and the annotation file into a Mask-RCNN network for training to obtain an organ recognition model; intercepting organs in the marked image from the original image, taking the organs and corresponding scores as training set data, and training different convolutional neural networks to obtain an image scoring model capable of scoring development of each organ; inputting a to-be-recognized mouse embryo original image into the organ recognition model, and outputting all organs in the image; intercepting all organs from the original image, and respectively inputting the organs into an image scoring model to obtain a development score of each organ; and obtaining a total score. According to the method, the organs in the mouse embryo can be quickly and accurately recognized, and the current development stage of each organ can be judged.

The invention discloses an improved method and a kit for detecting quality of an assisted reproductive technology by a mouse embryo array, and particularly relates to a method for detecting the quality of the assisted reproductive technology (ART) by utilizing cell number at each stage of mouse embryo development and UTF1 positive cell number expressed by blastocyst in the mouse embryo assay (MEA), belonging to the technical field of cytology and biology. At present, based on the internationally common standard MEA, the following two standards are increased: (1) total number of cells contained in embryo at each 12-hour embryonic development period; (2) number of UTF1 positive cells expressed by the blastocyst and number of CDX2 cells expressed by trophoblast cell as well as a proportion of the UTF1 positive cells and the CDX2 cells to total cells of the blastocyst. By comparing the two standards for in-vitro cultivated and developed embryo with the in-vivo embryo at the corresponding period, relatively sensitive and effective means are provided for quality control, including optimizing in-vitro culture conditions and optimizing embryo culture liquor, of the detected assisted reproductive technology, and reference standards are provided for improving the assisted reproductive technology.

The invention provides a novel body cell reprogramming inducing method, a kit and an application and relates to an application of an ectodermal differentiation and development correlation factor and / or mesendodermal differentiation and development correlation factor as an inducing factor to generation of an induced pluripotent stem cell. According to the invention, the inducing factor is provided for a differentiated stem cell to induce to generate the induced pluripotent stem cell (iPS cell), and the inducing factor comprises the mesendodermal differentiation and development correlation factor, Sox2, Klf4 and any one c-Myc or the ectodermal differentiation and development correlation factor, Oct4, Klf4 and any one c-Myc or the mesendodermal differentiation and development correlation factor, the ectodermal differentiation and development correlation factor, Klf4 and any one c-Myc. The iPS cell obtained by using the novel body cell reprogramming inducing method has characteristics similar to those of a rat embryonic stem cell, namely has self-renewal maintaining capacity and differentiation omnipotency.

The invention provides a method for obtaining a mouse model with neural tube defects (NTDs). The mouse model with the NTDs is established by applying an anti-inositol metabolism medicament to a pregnant mouse from the embryonic development of the pregnant mouse to the formation of neural tubes. The mouse model can be used for medical and biological research of the NTDs, is helpful to understand the causes and pathogenesis of human NTDs, and provides a model for further research on an action mechanism of inositol metabolic disorders in the NTDs. Moreover, according to the method, the neural tube defect rate is high, absorbed embryos are few, the embryo fatality rate is low, and toxic and side effects to the pregnant mouse are low.

The invention provides a method for establishing a heart disease drug screening model, which comprises the following steps of A) establishing a virus vector comprising four transcription factors: Oct3 / 4, Sox2, c-Myc and klf4; B) transfecting the virus vector in the step A) with a virus packaging cell to obtain virus liquid; C) infecting the virus liquid in the step B) with the fibroblast of the beta adrenergic receptor wild-type gene knockout mouse; D) screening the clone similar to the mouse embryonic stem cell, and subculturing to obtain the induced pluripotent stem cells with beta adrenergic receptor wild-type gene defect; E) establishing a virus vector containing the beta adrenergic receptor mutant gene; F) transfecting the virus vector in the step E) with the virus packaging cell to obtain virus liquid; G) infecting the virus liquid in the step F) with the induced pluripotent stem cells with beta adrenergic receptor wild-type gene defect; H) selecting positive clone and performing induced differentiation into myocardial cells.

The invention relates to the technical field of medicines, in particular to application of rapamycin in preparation of a miscarriage prevention medicine for spontaneous abortion. Compared with normalpregnancy, macrophages residing in decidua tissue of a patient suffering from recurrent spontaneous abortion are reduced, and expression of adhesion molecules is low; and the autophagy level of the decidual macrophages of the abortion patient is low. Experimental results show that the rapamycin can significantly promote adhesion of the macrophages to decidua matrix cells. Normal pregnant rats andabortion pregnant rat models are constructed, and in-vivo experiments prove that: the rapamycin is applied to reduce dwelling of decidual macrophages caused by autophagy disorders, maintains the steady state of decidua immune components and significantly reduces the embryo absorption rate of the pregnant rats, thereby improving the outcome of bad pregnancy. Therefore, the rapamycin is expected tobe used as the miscarriage prevention medicine for spontaneous abortion, a new treatment scheme is provided for patients suffering from repeated spontaneous abortion, the psychological burden of the patients is relieved, and the rapamycin can be well applied clinically.

The invention discloses application of UL16 protein in preparation of a small molecular medicine for promoting in-vitro differentiation of embryonic stem cells, and application of stem cell transplantation in treatment of nervous system degenerative diseases. According to the invention, by construction of UL16 plasmids to transfect host cells, the fact that UL16 can promote aerobic oxidation and oxidative phosphorylation of glucose and activate the cell productivity pathway so as to increase the intracellular ATP content and improve mitochondrial functions is found. Finally, by construction ofUL16 expression plasmids to transfect mouse embryonic stem cells, mitochondrial function state enhancement and energy metabolism enhancement are found, and stem cell differentiation can be promoted.Clinical treatment of cells in a differentiation defect state and cells with a low differentiation rate is realized; and the UL16 protein is used for treating related diseases or pathological and physiological processes of mitochondrial dysfunction or hypofunction of the cells.

The invention discloses a non-alcoholic fatty liver mouse model and a construction method thereof. The construction method includes the specific construction step that 1, mouse embryonic stem cells are transfected, and a fourth exon segment and a fifth exon segment containing ghr genes are amplified through Pfu Turbo DNA polymerase and cloned into a pCR Blunt II-TOPO vector. When a mouse modeled through the method is 8 weeks old, the liver volume is remarkably increased, H&E staining and oil red staining results show fatty liver lesion, liver cell fat synthesis is remarkably increased, lipolysis is remarkably reduced, and the mouse model built through the method has the advantages of being stable in heredity, similar to human conditions in clinical symptoms, short in modeling time, high in efficiency and low in death rate. The model is capable of being formed without diet induction, and relatively wide in application and popularization prospect, and is an ideal non-alcoholic fattyliver animal model for drug screening and pharmacological research.

The invention discloses a method for promoting differentiation of rat radial glial cells into neurons, comprising the following steps of: acquiring a fetal rat cerebral cortex cell suspension, and preparing RGCs single cells; and inoculating the RGCs single cells into a culture plate, culturing overnight by using an RGCs culture solution, and continuously culturing by using an RGCs induced differentiation solution. The method for preparing the RGCs single cells comprises the following steps of: resuspending cells by using an RGCs culture solution to form macroscopic suspended cell spheres, replacing the RGCs culture solution, and continuously culturing for 5-7 days to obtain the cell spheres; and treating the cell spheres with Accutase enzyme, digesting the cell spheres into single cells,continuing suspension culture, and carrying out passage. According to the method, the differentiation ratio of the rat embryo cerebral cortex derived radial glial cells to neurons can be obviously improved and can reach about 3 times or even higher than the natural differentiation rate.

The present invention discloses a method for improving induction efficiency of neural stem cells and promoting differentiation of neurons. A programmed induction of SMSINS cells is as follows: using LDN193189 (Selleck S2618, 0.25 [mu]M) and SB431542 (Selleck S1067, 1 [mu]M) on the 1st and 2nd days to treat mouse embryonic fibroblasts (MEFs); using CHIR99021 (Selleck S2924, 3 [mu]M) DAPT (Selleck S2215, 5 [mu]M) and VPA (Sigma P4543, 0.5 mM) on the 3rd and 4th day; using CHIR99021 (Selleck S2924, 3 [mu]M) and DAPT (Selleck S2215, 5 [mu]M) on the 5th and 6th days; adding Shh (Peprotech 315225, 100 ng / ml) and Purmorphamine (selleck S3042, 1 [mu]M) on the 7th and 8th days to improve neural differentiation; and removing all small molecule compounds on the 9th and 10th days and conducting changeto a NSC complete culture medium. At the time, SMSINS cells can be trypsinized into single cells and can be inoculated into 24-well plates without coating PDL / L in the NSC complete culture medium; and the SMSINS cells are subjected to in vitro neuronal differentiation. The method is simple and convenient to operate, can improve induction of the neural stem cells and promote the differentiation ofthe neurons, and is beneficial to popularization and application.

The invention relates to the usage of rat cardiac muscle cell cultured supernatant. The rat cardiac muscle cell cultured supernatant can be used for culture medium for proliferation with non-differentiation of rat embryonic stem cell and its preparation method. The rat cardiac muscle cell cultured supernatant can be used exclusively or by combination of auxiliary materials for rat embryonic stem cell special culture substrate. The preparation method for the culture substrate is also provided. To culture rat embryonic stem cell for proliferation without differentiation in vitro culture by means of the method can derive a better effect than leukemia inhibiting factor(LIF). The invention employs rat cardiac cell, has low cost and simple preparation process, being superior to LIF in promoting ES cell wall sticking, proliferating and cloning.

The invention belongs to the technical field of medicines, and in particular discloses application of fructose 1,6-diphosphate (FDP) in preparation of a fetus protection medicine for spontaneous abortion. In vitro and in vivo animal experiments show that the concentration of the FDP in peripheral blood and decidual tissues of patients with recurrent spontaneous abortion is decreased compared withnormal pregnancy. Compared with control pregnant mice, the model plasma and uterus FDP levels of pregnant mice with spontaneous abortion are reduced, the uterus macrophage M2 phenotype molecule expression of the pregnant mice with spontaneous abortion is decreased, the proportions of helper cells T(Th)2 and regulatory T cells are reduced, and the phenomena of poor endometrium decidualization and decreased embryonic trophoblast infiltration degree occur in an accompanied way. The results indicate that the supplement of the FDP can significantly improve the uterine immune tolerance pattern, decidualization and embryonic trophoblast infiltration in the pregnant mice with spontaneous abortion, and obviously reduces embryonic loss in the pregnant mice. Therefore, the FDP is expected to be an early-warning and diagnostic indicator for the patients with spontaneous abortion, and can be used for preparing the fetus protection medicine for the spontaneous abortion.

A process for constructing the vector pEGVPV, which can copy a lot of DNAs in procaryotic cell and is able to express in procaryon and mammal cell, includes such steps as cloning murinal VEGF-2 (mVEGF-2) gene from mouse embryo, cloning human VEGF promoter, linking them and inserting the gene into mammal's expression carrier pEGSH. After said carrier is used to transform E. coli DH5 alpha, the output of DNA separated from the culture medium of cells can reach 1.5 mg / L. A process for preparing the vascular endothelial growth factor 2 as DNA medicine from microbe for treating vascular diseases is also disclosed.

The invention discloses a method for obtaining the submandibular gland of mouse embryos, and relates to the field of biomedicine. The method includes: freeing the uterus: after killing the pregnant mouse, freeing the uterus containing the embryo completely; freeing the embryo: opening the uterine wall, freeing the embryo completely; obtaining embryonic jaw tissue: obtaining the jaw tissue from the embryo under a dissecting microscope ;Obtain embryonic submandibular gland: place the mandibular tissue under a dissecting microscope, zoom in on the field of view clearly and then fix it. After separating the most superficial tissue, pinch off the connection between the submandibular gland, the tongue body and the duct, and then obtain the complete embryonic submandibular gland. This method improves the operation of obtaining the submandibular gland of mouse embryos. Compared with the traditional method, the operation steps are reduced, the operation difficulty is reduced, and the integrity of the extracted submandibular gland tissue is guaranteed to a greater extent, thus providing a better method. The model of submandibular gland cultured in vitro provides a reliable platform for the research of submandibular gland related diseases.

The invention discloses a method for rapidly and directly directionally inducing the differentiation of mouse embryonic stem cells into neuroepithelial cells, by adding an induction medium containing dorsomorphin and noggin, SB431542 and CHIR99021 to the adherent cultured mouse embryonic stem cell pellet clones to induce mouse Embryonic stem cells form rosette-shaped neurospheres; then add the second-stage induction medium containing bFGF to form suspended neurospheres; finally digest the neurospheres into single cells, and then use the third-stage induction medium to form adherent cultures Neuroepithelial cells with longer self-proliferation and renewal. The invention abandons the traditional EB forming method, shortens the differentiation time, and the differentiation efficiency reaches more than 95%. The invention can quickly induce NE cells to replace NSC cells to treat central system damage, and has high clinical applicability.

The invention relates to the technical field of medicine, and provides a method of screening and identifying small molecule with anti-aging properties. The method comprises cellular models, namely premature aging mouse embryonic fibroblasts and human HGPS mesenchymal stem cells and animal models, namely premature aging progeroid mice and Caenorhabditis elegans. The method successfully screened and identified Cannabidiol (CBD), which had senolytic effects. The invention also determined that the optimal concentration of CBD treatment is 10-20 μM.

The present disclosure is directed to the development of compositions, such as extracellular matrices, and processes for using the same, for culturing stem cells in vitro in an undifferentiated state. In this regard, it has been discovered that when pluripotent mouse and human embryonic stem cells are cultured on plates coated with recombinant laminin-10 (laminin-51 1 ) or laminin-5 (laminin-322), or their functional domains, the embryonic stem cells proliferated and maintained their pluripotency.

The invention provides an application of a polymethyl methacrylate material in manufacturing a CUBIC transparent tissue microscopic imaging holder. The invention also provides a method for preparing the CUBIC transparent tissue microscopic imaging holder, which comprises the following steps: adding a polymethyl methacrylate material into a blow molding device, an injection device or an extrusion device for molding. The polymethyl methacrylate material can be reshaped into holding devices with various shapes and sizes after being heated, and is suitable for various biological tissue samples after CUBIC transparency, such as zebra fish, mouse embryo, brain and human tumor tissues. The holder prepared from the material disclosed by the invention can be fixedly transferred in a light sheet microscope sample bin, and a sample to be shot can be quickly and conveniently placed in the holder, so that the transparent sample can be imaged more completely and conveniently.

The invention belongs to the field of medicinal chemistry, and discloses a compound with a benzomorpholone-biphenyl skeleton as well as a preparation method and application thereof. The compound has a structural general formula (I) as shown in the specification. The preparation method of the compound is simple and reliable, and the total yield is more than 40%. Preliminary pharmacological experiments show that the compound has a certain relaxation effect on in-vitro artery blood vessels of rats, can inhibit proliferation of mouse embryo fibroblasts (NIH-3T3) and expression of type I collagen to a certain extent, and can be used for development of anti-hypertension drugs and anti-myocardial fibrosis drugs.

The invention belongs to the field of artificial intelligence, and specifically relates to a mouse embryo organ recognition and scoring method and system. The method includes the following steps: collecting original images of mouse embryos in different periods and manually labeling them; Input the Mask-RCNN network for training to obtain the organ recognition model; intercept the organs in the marked image from the original image, and use it and the corresponding score as the training set data, and train different convolutional neural networks respectively. Obtain an image scoring model that can score the development of each organ separately; input the original image of the mouse embryo to be recognized into the organ recognition model, and output all organs in the image; intercept all organs from the original image, and input them into the image scoring model Model, get the development score of each organ; get the total score. The invention can quickly and accurately identify the organs in the mouse embryo, and judge the current development stage of each organ.

The invention relates to a mouse embryonic stem cell strain SXMUe002-A-1, which has a preservation number of CGMCC No: 23035, and is preserved in the China Center for Type Culture Collection on September 28, 2021. The invention also provides an application of the mouse embryonic stem cell strain SXMUe002-A-1 in preparation of a mouse hemophilia model. According to the obtained mutant cell strain SXMUe002-A-1, the growth and reproduction capability and the cellular morphology of the mutant strain are highly similar to those of a wild type, no base is deleted and inserted on an exon, only one base is mutated, and the mutant cell strain SXMUe002-A-1 is closer to clinical cases. The mutant cell strain SXMUe002-A-1 has the characteristics of mouse embryonic stem cells, can be subjected to mouse embryonic stem cell chimeric injection to obtain a hemophilia mouse model, and is of great significance in further hemophilia nonsense mutation mechanism research, hemophilia drug screening and gene therapy.

A tool for mouse embryo transplantation preservation and its manufacturing method, comprising a centrifuge tube, a syringe, a rubber piston and a Pasteur tube, the rubber piston is inserted into the syringe jacket from the tail, and a part of the volume of the cone head of the syringe jacket is appropriately cut off to make the cone The width of the mouth is equal to the width of the outer diameter of the Pasteur tube. Properly cut off the two wings of the syringe jacket so that the length of the two wings is equal to or slightly greater than the diameter of the centrifuge tube. Insert the syringe jacket into the centrifuge tube and fix the jacket so that the jacket cannot be left or right in the centrifuge tube. Moving up and down, the Pasteur tube is fixed to the cone of the jacket. By adopting the invention, the transplanted embryos can be effectively preserved in the external environment, shortening the low temperature and low humidity time of the embryos in vitro, and facilitating the implantation and later development of the embryos.

The invention discloses an albumin / keratin double expression mice embryonic stem cell system and an establishing method thereof. The mice embryonic stem cell system of the invention is established by sub-culturing the albumin / keratin double expression mice embryonic stem cell. The mice embryonic stem cell is integrated with a red fluorescence / green fluorescence protein double reporter gene, wherein the reporter gene is regulated by tissue specificity promoter albumin ALB or keratin CK19. Compared with a method of inputting genes, the method of controlling the reporter gene by using the transfection tissue specificity promoter of the invention is more simple, rapid and useful. The uniform originator cells in the developmental stage can be precisely chosen by using the albumin / keratin double expression mice embryonic stem cell of the invention, which can bring a great beneficial effect to the following research on the originator cell differentiation, the cell transplanting and cure.

The invention provides a novel method, a kit and an application for introducing somatic reprogramming. The invention relates to an application of correlation factors for mesendoderm differentiation and development as inducing factors for producing induced pluripotent stem cells. The inducing factors are provided to differentiated cells so as to induce and produce the induced pluripotent stem cells (iPS cells), wherein the inducing factors includes the correlation factors for the mesendoderm differentiation and development, Sox2, Klf4 and optionally c-Myc. The iPS cells produced by using the correlation factors for the mesendoderm differentiation and development to replace Oct4 have similar characteristics with mouse embryonic stem cells, that is, the iPS cells have totipotency for maintaining self-renewal capability and differentiation.

The invention discloses an application of andrographis paniculata flavonoid glycoside C in preparation of a medicine for preventing and treating Covid-19. A BEAS-2B human normal pulmonary epithelial cell model, an NIH-3T3 mouse embryonic cell model, a molecular docking technology and molecular dynamics simulation are adopted to prove that the andrographis paniculata flavonoid glycoside C has no obvious toxicity on the human normal pulmonary epithelial cell and the mouse embryonic cell; the compound has a strong affinity effect and an enzymatic activity inhibition effect on angiotensinase 2 on human normal pulmonary epithelial cells. The andrographis paniculata flavonoid glycoside C can be combined with human lung epithelial cells ACE2 to block the combination of the SARS-CoV-2 and the ACE2, so that the invasion path that the SARS-CoV-2 depends on the combination with the ACE2 to enter the cells is inhibited, and the effect of preventing and treating Covid-19 is achieved. By researching the cytotoxicity of the andrographis paniculata flavonoid glycoside C and the affinity and inhibitory activity of the andrographis paniculata flavonoid glycoside C to human lung epithelial cells ACE2, a new thought and scheme are provided for research, development and production of medicines for preventing and treating Covid-19.

The invention discloses a culture medium for epitaxy culture of a mouse embryo body, which contains a basic culture medium, fetal calf serum, human umbilical serum, sodium pyruvate, glutamine, penicillin-streptomycin, an N2 additive and a B27 additive, and can promote the continuous culture stage of an in-vitro mouse embryo to E8.0 on the basis of improving the culture success rate. A foundation is provided for development research of pre-implantation embryos and implantation stage embryos, the selection range of mouse embryo related experiment contents and experiment stages can be expanded by improving the culture success rate and prolonging the in-vitro culture time, and a basis can also be provided for in-vitro culture of other mammal embryos.

The invention relates to the technical field of culture of KM mouse embryos, in particular to an observation device for culture in vitro of KM mouse embryos. The observation device comprises an observation box, wherein an observation mirror is mounted at the top end of the observation box, first support rods are welded to four corners of the bottom end of the observation box, a fixing block is inclamping connection to the top ends of the first support rods, a first clamping groove cooperating with the corresponding first support rod is formed in the bottom end of the fixing block, a placing plate is welded to the top end of the fixing block, second clamping grooves are uniformly formed in the top end of the placing plate, a first clamping block is in clamping connection to the inner partof the corresponding second clamping groove, a sterilizing protection pad is bounded to the top ends of the first clamping blocks, a switch gate is in hinged connection to the peripheral wall body ofthe observation box, above the placing plate, and a plurality of observation windows are mounted at the middle of the switch gate. According to the observation device, the placing plate can be convenient to disassemble, so that the placing plate can be conveniently cleaned; and observation windows and the observation mirror are arranged, so that comprehensive observation of the KM mice can be realized.

The invention relates to an experimental method for inducing directional differentiation of neural stem cells into neuron cells by Gd:Fe3O4@RA nanoparticles. The experimental method includes the following steps of putting a cover slip pre-treated with polylysine into a culture plate, and inoculating culture holes with primary and passaged neural stem cells derived from rat embryonic brain hippocampus at 1 is multiplied by 10<7> L<-1>, removing basic fibroblast growth factors from a complete culture solution of the neural stem cells, adding 1.0 ml of Gd:Fe3O4@RA complex nano ions with the concentration of 1 mg / ml and fetal bovine serum with the volume fraction of 5% to promote neural stem cell differentiation, performing culture for 6 days, continuing culture for 6 days in a neural stem cell culture medium containing the fetal bovine serum with the volume fraction of 5%, and taking out the cover slip after the cell morphology is mature.