1443results about "Embryonic cells" patented technology

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention discloses an artificial breeding method of epinephelus fuscoguttatus and pink pterus coerulescens hybrid species, which comprises the following steps: screening healthy epinephelus fuscoguttatus as a female parent, screening healthy pink pterus coerulescens as a male parent, and carrying out enhanced breeding on the male parent and the female parent, the gonad maturation of the parent is promoted; the method comprises the following steps: collecting egg liquid of a female parent and semen of a male parent, inseminating to obtain a fertilized egg, incubating the fertilized egg to obtain a hybrid spot, performing authenticity identification on the hybrid spot by using one or more of sequences SEQ ID NO.1-SEQ ID NO.8, and determining that the hybrid spot inherits genetic information of the female parent and the male parent. The technical blank of crossbreeding of the pink pink bass as the parent grouper is filled, so that the hybrid obtained by hybridizing the epinephelus fuscoguttatus and the pink pink bass is reliable in authenticity, and inherits genetic information of male and female parents on the molecular level.

The present invention relates to a modified stem cell and an ex vivo method for preparing defined mature cells from genetic programming of said modified stem cell which can be carried out in absence of certain growth factors, as well as a kit allowing to transform stem cells, and the different uses of the mature cells, for example for in vitro drug screening and toxicology or as cultivated meat. The genetic programming involves a molecular switch between two mutually exclusive programs of stem cell expansion and differentiation. The invention may be used for stem cells or other progenitors cells of any type, from any eukaryotic organism, but finds particular application in pluripotent stem cells from humans and livestock animals.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a Myo1B protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and / or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ / Activin / Nodal signaling and BMP signaling are provided. Also provided are methoc and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some embodiments, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some embodiments, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

The invention relates to a method for inducing mouse trophoblast stem cells in vitro. According to the method, mouse embryonic stem cells are used as starting cells, Sorcs3 gene knockout mouse embryonic stem cells are obtained through a gene editing technology, and in-vitro induced trophoblast stem cells (iTSCs) are efficiently and rapidly obtained by means of a flow cytometry sorting technology and in combination with specific antibody sorting. According to the invention, the Sorcs3 gene is knocked out from the embryonic stem cell, so that the capability of the embryonic stem cell to differentiate to the trophoblast lineage can be obviously enhanced; an important research platform is provided for deeply researching a lineage intertransformation mechanism of cell masses (ICM) and trophoblast (TE) in the blastocysts, and the method has important significance for researching lineage limitation of early embryonic development of mammals and the like.

The invention provides hindbrain organoids and methods of producing such organoids. Said hindbrain organoids may include healthy or healthy and diseased tissue. Also provided are methods of testing therapeutic agents using said hindbrain organoids. The invention also provides culture mediums used for producing (patterning) said hindbrain organoids and kits including the culture medias.

The present invention relates to the new use of an exosome secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and in particular relates to the use of the exosome in the preparation of a drug for treating liver diseases. The exosome is secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and the hepatocyte lineage cells are hepatic progenitor cells and / or mature hepatocyte-like cells.

The invention relates to a new technology for promoting generation of hematopoietic precursor cells from human pluripotent stem cells. Specifically, the present invention relates to a method for preparing hematopoietic precursor cells comprising reducing the expression level of YTHDF2 gene in pluripotent stem cells or reducing the activity of YTHDF2 protein of pluripotent stem cells. On the other hand, the invention also relates to a kit for preparing the hematopoietic precursor cell, and the kit comprises a reagent for reducing the expression level of the YTHDF2 gene or a reagent for reducing the activity of the YTHDF2 protein.

The invention provides a method for evaluating drug enterotoxicity in vitro through an intestinal organ model, and relates to the field of biological medicine. Comprising the following steps: preparing six different culture media, culturing the hESC by using the different culture media, inducing the hESC to respectively perform directional endoderm cell differentiation, posterior intestinal bulb differentiation, intestinal organ differentiation and intestinal organ maturation, and evaluating the drug enterotoxicity by using a prepared intestinal organ model. The intestinal organ model prepared by the invention is similar to human intestinal cells in composition, can accurately reflect the toxic effect of the medicine in the physiological environment in the human body, can be repeatedly obtained in quantity, and avoids clinical risks and ethical constraints; the sensitivity of drugs with different enterotoxicity can be reflected through cell viability, and the method is suitable for rapid evaluation of in-vitro high-flux drug enterotoxicity.

The invention provides a serum-free stem cell culture medium and a preparation method thereof, and the culture medium realizes long-term stable culture of stem cells under a completely serum-free condition by optimizing basic culture medium components and adding a specific growth factor combination, an extracellular matrix simulant and a metabolism regulator. The culture medium comprises a basic culture medium, a recombinant human protein substitute, a cell adhesion promoting factor, a growth factor combination, an antioxidant and a metabolism regulator. Compared with the prior art, the invention has the following advantages: 1) animal-derived components are completely avoided, and the immunogenicity and the difference between batches are reduced; 2) maintaining the dryness of the stem cells through a specific growth factor combination; 3) adding a metabolism regulator to optimize cell energy metabolism; 4) the cost is obviously lower than that of commercially available like products; and 5) supporting long-term culture of stem cells without spontaneous differentiation. The invention also provides a preparation method of the culture medium and a method for culturing stem cells by using the culture medium.

Disclosed herein are methods of producing organoids. In some aspects, the methods involve producing organoids by implanting primary cancer cells such as glioblastoma cells into brain organoids, thereby producing brain cancer organoids. The method can comprise an incubation step in which the brain organoid is incubated with primary cancer cells in a hanging liquid droplet to produce the brain cancer organoid. In some aspects, the brain cancer organoids provide tools for understanding and treating cancers of the nervous system, such as glioblastoma.

The present disclosure relates to organoids and particularly to brain organoid models. The brain organoids include neurons, microglia, astrocytes, and blood vessels. The brain organoid models can be used to create models for Alzheimer's Disease. Methods of using the brain organoids for drug discovery are also described.

The present invention discloses a system for targeted gene editing and related uses. Also disclosed are related cells.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a glucose slow-release structure used in an embryo in-vitro culture solution, a preparation process and a use method. The glucose slow-release structure comprises at least one inner layer containing glucose and a biodegradable material outer layer wrapping the inner layer, and the biodegradable material outer layer comprises a matrix with a plurality of micropores and at least one sealing material layer used for sealing pores of the micropores. According to the invention, gradient release from low sugar in the cleavage stage to high sugar in the blastocyst stage is realized, midway liquid change or artificial sugar supplement is not needed, the operation is obviously simplified, and pollution and mechanical stress risks are reduced.

The invention discloses a vector system and method capable of converting mouse embryonic stem cells into embryonic-like cells in a 2-cell period, the vector system comprises a mouse Zfp352 gene sequence, and the nucleotide sequence of the mouse Zfp352 gene is shown as SEQ ID No.1. The invention also discloses a method for converting the mouse embryonic stem cells into embryonic-like cells in a 2-cell period. The invention creatively discovers that through overexpression of the Zfp352 in the mouse embryonic stem cells, the efficiency of converting the mouse pluripotent embryonic stem cells into the totipotent embryonic-like cells in the 2-cell period can be remarkably improved.

The present invention aims to provide a method for efficiently producing a cell mass containing pituitary tissue from pluripotent stem cells. A method for producing a cell mass containing pituitary tissue, including the following steps (1) and (2):(1) a first step of suspension-culturing pluripotent stem cells to form a cell aggregate in the presence of a Wnt signal transduction pathway inhibiting substance,(2) a second step of suspension-culturing the aggregate obtained in the first step in the presence of a BMP signal transduction pathway activating substance and a Sonic hedgehog signal transduction pathway activating substance, thereby obtaining a cell mass comprising pituitary tissue.

The present invention provides a method of producing a population of mesenchymal stromal cells (MSC) of CD73 + CD44 +, CD90 +. The CD73 + CD44 +, CD90 + MSC is used in a method for producing terminally differentiated osteoblasts, adipoblasts and chondroblasts from pluripotent stem cells (PSCs). The differentiation methods include the use of a single agent-a WNT signaling pathway activator used on an adherent culture of PSC, such as a GSK3 [beta] inhibitor.

The present application relates to a kind of RIC cells and its preparation method and application, especially it is related to the lineage-specific differentiation of pluripotent or multipotent stem cells, specifically related to a kind of RIC cell obtained from stem cell differentiation and its preparation method and application.

Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells and a culture medium composition thereof. Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells or NK cells and a culture medium composition thereof. The NK cells obtained using the provided culture method have high purity and good in vitro expansion effect; the yield of iNK cells is high, and a single iPSC can differentiate into approximately 2,000 NK cells; and the obtained iNK cells highly express CD16 (over 70%). As a result, the problem of low CD16 expression in iNK cells in the prior art, which requires genetic modification methods to solve, has been addressed.

Disclosed are methods for producing controlled endometrial organoids using micropatterning techniques, and methods for using endometrial cells or organoids for diagnostic and predictive purposes, and for optimizing IVF and other infertility-related treatments, including embryo transfer and intrauterine insemination (IUI). These organoids grow in a spatially controlled manner and exhibit less heterogeneity than organoids grown directly in a three-dimensional culture. The specification also describes methods for predicting the success of embryo transfer or other IVF-related treatments, optimizing embryo transfer, improving embryo implantation in the endometrium, and performing embryo transfer in subjects. These methods include culturing endometrial organoids or endometrial epithelial cells, and contacting the endometrial organoids or cells with one or more blast-like cells. In some embodiments, the endometrial organoids or cells include endometrial cells obtained from subjects undergoing or considering IVF-related treatment.

This disclosure relates to cell therapy approaches for treating progranulin (PGRN) deficiencies with human induced pluripotent stem cell (hiPSC)-derived cells. Advantageously, the hiPSC-derived cells (e.g., microglia progenitor cells) described herein can cross-correct PGRN deficiencies in damaged or diseased cells while reducing the amount of endogenous cell ablation that is needed, as demonstrated by experimental results showing restoration of PGRN levels in GRN mutant cells and brain organoids.

The invention belongs to the field of stem cell culture, and relates to a pluripotent stem cell culture medium, which comprises an E6 basal culture medium, 2 ng / mL of TGF-beta1, 0-50 ng / mL of FGF-2, and 0.01-1 [mu] M of SUN11602. According to the technical scheme, proliferation and pluripotency of the hPSC are effectively maintained in a mode of combining small chemical molecules with low-concentration FGF-2.

To provide a composition useful for culturing and differentiating pluripotent stem cells.SOLUTION: A composition comprising cultured radial glial-like cells expressing at least one marker of Brain Lipid-Binding Proteins (BLBP), CD133 (Prominin 1), Abnormal Spindle Microcephaly Associated Proteins (ASPM), Baculovirus Inhibitors of Apoptosis Repeat Containing 5 (BIRC5 or Survivin), FAT atypical cadherin 1 (FAT1), Hes family bHLH transcription factors 5 (HES5), SRY-Box transcription factors 21 (SOX21), or PAX6 proteins, A composition wherein the radial-glial-like cells are or have been cryopreserved in a cryopreservation medium comprising Chroman1 and / or a derivative thereof, Emricasan and / or a derivative thereof, trans-ISRIB and a polyamine comprising putrescine, spermine and spermidine.SELECTED DRAWING: None

A sequential cell culture system and methods for the in vitro generation of human spermatozoa from differentiating spermatogonia (dSPGs) and spermatogonial stem cells (SSCs) is provided. The instant disclosure encompasses the discovery and optimization of distinct, stage-specific culture media and conditions that enable, for the first time, the complete progression of human spermatogenesis outside the body. The process includes: (1) inducing entry of dSPGs and SSCs into meiosis; (2) supporting meiotic progression and completion to round spermatids; and (3) promoting spermiogenesis and the formation of elongated, spermatozoa. The invention further provides compositions, systems, and protocols for each stage, as well as markers and methods for identifying successful progression through each developmental transition. This system enables, for the first time, the complete in vitro recapitulation of human spermatogenesis, providing a platform for research, infertility treatment, and assisted reproductive technologies.