109 results about "Mouse Embryonic Stem Cell" patented technology

The invention discloses a method for producing transgenic mice through a homologous recombination technology. The method comprises the following steps: replacing mouse NGF genes on mouse chromosomes by human NGF genes through a Cas-9/CRISPR gene knock-in technology, and obtaining the genes with homozygous human NGF genes through breeding and knocking the genes in mice, thus obtaining filial generation mice with salivary glands capable of secreting human NGF. Compared with the conventional targeting technology utilizing positive and negative double screening homologous recombination genes, the method disclosed by the invention is simple to operate, capable of being realized only by transfecting mouse embryonic stem cells by three plasmid DNAs or carrying out pronucleus injection on the zygotes of mice, and high in success rate which is up to 2-5% and remarkably higher than the mouse embryonic stem cell positive rate of 0.1% of the common gene targeting technology.

The present invention relates generally to the field of cell biology of stem cells, more specifically the directed differentiation of pluripotent or multipotent stem cells, including human embryonic stem cells (hESC), somatic stem cells, and induced human pluripotent stem cells (hiPSC) using novel culture conditions. Specifically, methods are provided for obtaining neural tissue, floor plate cells, and placode including induction of neural plate development in hESCs for obtaining midbrain dopamine (DA) neurons, motorneurons, and sensory neurons. Further, neural plate tissue obtained using methods of the present inventions are contemplated for use in co-cultures with other tissues as inducers for shifting differentiation pathways, i.e. patterning.

The invention relates to application of inorganic nano-material layered double hydroxides (LDHs) in mouse embryonic stem cell culture. Under the condition that LIFs are not added, the inorganic nano-material LDHs can promote expression of pluripotency genes and restrains cell differentiation, and in addition, the treated cells still have the potency of differentiating to three germ layers. The inorganic nano-material LDHs have excellent biocompatibility, low cytotoxicity, easiness in contact with cell surfaces, low cost, convenience and rapidness in material synthesis and excellent effect, and has extensive application prospect in research of embryonic stem cells.

Differentiation of human pluripotent stem cells, such as human embryonic stem cells (hESC), into hepatocytes by in vitro methods is disclosed. The pluripotent stem cells are cultured in conditioned medium from the hepatocarcinoma cell line, HepG2. Specific growth factors and defined media may also be added to the medium for stage specific differentiation of the derived hepatocytes. Hepatocytes differentiated from human pluripotent stem cells may be characterized by fluorescence activated cell sorting (FACS), immunofluorescence analysis (IF), real time polymerase reaction (RT-PCR), and functional assays. The methods disclosed herein are able to differentiate high percentages of hepatocytes from human pluripotent stem cells using the disclosed methods. These differentiated cells may exhibit polygonal shape morphology, typical of hepatocytes, and may express hepatocyte specific genes. The differentiated cells may also be positive for definitive endoderm markers and hepatic markers.

The invention belongs to the technical field of medicines, and particularly relates to a myocardial small molecular compound for idiosyncratically promoting proliferation of myocardial cells and application thereof. The myocardial small molecular compound total comprises a type A, a type B and a type C which have a common core structure, namely (3 (1, 2, 4) triazole (3, 4-b) (1, 3, 4) bismuththiol). The myocardial small molecular compound is obtained from a small molecular compound library by means of model organism zebra fish screening, so as to be capable of idiosyncratically promoting proliferation of myocardial cells inside zebra fish bodies. Importantly, the myocardial small molecular compound is capable of inducing embryonic stem cells of a small mouse to differentiate and proliferate towards the myocardial cells. As proved, the myocardial small molecular compounds are antagonists to inhibit wnt signaling pathways. The myocardial small molecular compound can be used as pilot drugs for treating heart diseases and further is used for optimizing and evaluating candidate drug targets thereof.

The invention relates to a method for inducing human embryonic stem cells to be differentiated into retinal pigment epitheliums (RPE) in vitro, and belongs to the biology technical field. The pure RPEs can be obtained through one-time passage by means of induced differentiation. Compared with the prior art, the differentiation time from the human embryonic stem cells into the RPEs is shortened. By the adoption of the method, a new approach and guidance are provided for induced directional differentiation of the human embryonic stem cells into the pigment epitheliums.

The invention discloses a target targeting vector, a method for constructing a mouse embryonic stem cell strain through targeting integration of exogenous genes to an MYH9 Intron2 locus and application. The sequence of the target targeting vector is shown as SEQ ID NO.7, and the target targeting vector is named an mpNTKV-LoxP MAI2L-MAI2R vector. A new locus sequence capable of achieving efficient, safe and targeting integration is provided. According to the targeting vector constructed based on the locus and inserted into the exogenous genes in a targeting mode, efficient targeting can be achieved, it can be guaranteed that the exogenous genes are stably expressed in mouse embryonic stem cells and endogenous gene expression is not influenced, and therefore a good platform is erected for construction of a transgenic cell line and a rat.

The invention provides inducible CRISPRon or CRISPRi mouse embryo stem cells and a preparation method thereof. The mouse embryo stem cells are A2Lox.Cre cells and can reversibly express dCas9-VPR fusion protein or dCas9-KRAB fusion protein. The invention further provides a method for regulating and controlling gene expression and a kit. According to the inducible CRISPRon or CRISPRi mouse embryo stem cells provided by the invention, the construction method is simple and convenient, and the stable positive rate is up to 90 percent or higher; the genomic sequence is not edited, and gene expression is directly activated or inhibited; expression of the fusion protein has inducibility and reversibility; controllability and diversity on targeted regulation and control on the gene are achieved.

The invention discloses a method for quickly, directly and directionally inducing differentiation from mouse embryonic stem cells to neuroepithelial cells. The method comprises the steps of adding an induction medium containing dorsomorphin, noggin, SB431542 and CHIR99021 to a bulk clone of the mouse embryonic stem cells subjected to adherent culture, to induce the mouse embryonic stem cells to form rosette neurospheres; then adding a second stage of induction medium containing bFGF and performing suspension culture to form suspended neurospheres; and finally, digesting the neurospheres into single cells, and then performing adherent culture with a third stage of induction medium to form the neuroepithelial cells with long-term self proliferation and update. By adopting the method, the traditional EB method is abandoned, the differentiation time is shortened, and the differentiation efficiency reaches over 95%. The method can be used for quickly inducing NE cells to substitute NSC cells to treat central system injury, and has very high clinical applicability.

The invention discloses a mouse embryo stem cell culturing method and specific cultivating base, which cultures embryo stem cell on the trophoblast cell, wherein the trophoblast cell is epithelial cell or endothelium cell; the cultivating base consists of epithelial cell or endothelium cell and culturing liquid of embryo stem cell, which is high-sugar DMEM with 10-20% embryo cow serum or calf serum with 1. 5-3. 0*10-4mol/L Monothioglycerol, MTG.

The present invention discloses a series of alkylamino BODIPY dyes, methods for preparing a library of alkyl-amino BODIPY dyes via solid-phase synthesis, and the use of the alkyl-amino BODIPY dyes as fluorescent sensors for protein detection, cell imaging and cytometry applications, and staining of certain cell line.

The invention discloses a method for inducing differentiation of mouse embryonic stem cell to obtain inner ear hair cells, which comprises the following steps: using a conditioned medium and an induction nutrient solution for inducing embryonic stem cell for differentiation of precursor cells of hair cell; and then using a combination method of the induction nutrient solution and trophoblastic cells for inducing precursor cell for differentiation of mature inner ear hair cells. The method can effectively induce the differentiation of embryonic stem cell to obtain the precursor cells of inner ear hair cell and mature inner ear hair cell, the differentiation ratio is high, hair cell specific gene and protein can be expressed by cells obtained through differentiation, and the cells have certain electrophysiological function of hair cells.

This invention provides compositions for differentiating an isolated embryonic stem cell or an isolated embryoid body into neuronal progenitor cells and methods for using same.

The invention provides a chemical embryotoxicity prediction model. The chemical embryotoxicity prediction model comprises a cell type. The cell type comprises myocardial cells induced and differentiated from SP3ES cells with the karyotype as XX. An establishing method of the chemical embryotoxicity prediction model comprises the steps that 1, mouse embryo fibroblasts are separated and cultured; 2, in vitro multiplication culturing of the mouse ESs is conducted; thirdly, the karyotype analysis of the mouse ESs and gender determination are conducted; fourthly, the pluripotency of the mouse ESs is maintained and evaluated; fifthly, the mouse ESs are induced and differentiated to be myocardial cells in a in-vitro mode; sixthly, chemoimmunology detection of a cardiac-specific marker protein of the myocardial cells induced and differentiated from the mouse ESs is conducted; seventhly, real-time PCR detection of the expression profile of the myocardial cells induced and differentiated from the mouse ESs is conducted; eighthly, a pattern chemical is selected; ninthly, the cytotoxicity of the pattern chemical is detected; tenthly, the restraining effect of the pattern chemical on the ES differentiative capacity is detected; eleventhly, the embryotoxicity of the pattern chemical is evaluated. The chemical embryotoxicity prediction model fills up the blank that female mouse embryonic stem cells do not exist in an EST system.

The present invention belongs to the technical field of biology and discloses a targeted targeting vector, and a method and an application for targetedly integrating a foreign gene to a 22nd positionof a mouse F4/80 exon to construct BAC clone. A sequence of the targeted targeting vector is shown as SEQ ID NO.14 and named as a vector F4/80 5HA-IRES-DTR-PGK-EM7-Neo-F4/80 3HA. The present inventionprovides a new site sequence capable of achieving targeted integrating, the site can be used to construct the targeting vector for inserting the foreign gene, the targeting vector is verified to be capable of efficiently integrating the foreign gene into the mouse F4/80 BAC clone F4/80 exon 22th site, and the subsequently obtained targetedly inserted foreign gene F4/80 BAC can be used to construct mouse embryonic stem cells with the targetedly inserted foreign gene, thereby establishing a foundation for constructing a transgenic cell line and mice.

The invention discloses a method for inducing an embryonic stem cell into pancreatic tissue-like cells. The method comprises the following steps of: using a hanging-drop preparation method to differentiate an embryonic stem cell into three germ layer cells and using a immunomagnetic bead cell sorting method to purify finalized inner germ layer cells; further inducing the finalized inner germ layer cells so as to differentiate the cells into pancreatic precursor cells; and further differentiating the pancreatic precursor cells outside the body so as to form pancreatic tissue-like cells with high amylase secretion capability. The technical scheme of the invention adopts a plurality of induction factors for induced differentiation in different stages, which proves that the embryonic stem cell has the potentiality of being differentiated into pancreatic exocrine cells, and an effective way for inducting the embryonic stem cell into pancreatic tissue-like cells is found out, so that the pancreatic precursor cells are possible to recover injury of pancreas, and more cell sources are provided for recovering pancreas.

The invention provides a method for establishing embryonic stem cells containing exogenous chromosome. Specifically, the method injects a single chromosome inserted into a mCherry reporter gene into mouse fertilized egg by a microinjection mode, so that the mice embryonic stem cells (ESCs) containing exogenous chromosomes is produced. The method of directly transferring modified chromosomes by single-chromosome microinjection to mediate chromosome engineering is a very effective technique, and can promote the acquisition of cells containing the exogenous chromosomes and chromosome operation.

Methods and agents useful for modulating histone proteolysis, stem cell differentiation, and gene transcription and for treating cancer are disclosed. Antibodies or antigen binding fragments that selectively bind to histone-3 cleavage products and are useful for diagnosing cancer and monitoring a subject's response to cancer treatment are also disclosed.

The invention discloses an oriental breeding and differentiating method of stem cell in the stem cell tissue engineering domain, which comprises the following steps: adopting sodium alginate and polylysine as raw material; preparing APA mouse microencapsulation embryo stem cell; transplanting mouse microencapsulation embryo stem cell into different anatomical positions of living body orientally; realizing oriental breeding and differentiation of stem cell. The invention simplifies the operation, which is fit for oriental transplantation, recycling and internal oriental inducing differentiation study in the stem cell body.

The invention discloses an application method of small molecules capable of promoting self-status-renewal of embryonic stem cells. The application method of the small molecules capable of promoting self-status-renewal of the embryonic stem cells comprises the following steps: passaging and culturing human and mouse embryonic stem cells under small molecular state of the method; and then, observingand detecting self-renewal state of the cells according to the method. Being added with CID755673 small molecules, the application method of the small molecules capable of promoting self-status-renewal of embryonic stem cells can solve the problems of liable state differentiation, complex passages operation, high culture-condition cost, unfavorable follow-up mechanism research and so on of humanand mouse embryonic stem cells under existing traditional cultivation conditions; moreover, the application method of the small molecules capable of promoting self-status-renewal of embryonic stem cells has the advantages of being stable in effects, economical, highly efficient, easy to operate, convenient to study and so on. In addition, clues are further provided for improving and establishing culture conditions of pluripotent stem cells of other species and genera; and thus, the application method of the small molecules capable of promoting self-status-renewal of embryonic stem cells playsan active role in promoting smooth development of basic and applied research of stem cells in the future.

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.