241 results about "Cells embryo" patented technology

Methods and cell culture medium for the generation of human pluripotent embryonic stem cells are disclosed. Human embryonic stem cells are cultured with human granulosa feeder cells, muscle cells, Fallopian ductal epithelial cells, bone marrow stromal cells, and skin fibroblasts and the embryonic stem cells maintain their pluripotent phenotype. The human pluripotent embryonic stem cells can be cultured without feeder cells, and in the presence of supplemental growth factors. The human pluripotent embryonic stem cells can be alternatively cultured with conditioned medium obtained from a cell culture capable of maintaining human embryonic stem cells in a pluripotent state, wherein the cell culture is a human granulosa cell culture.

The invention relates to methods that allow for the efficient differentiation to form pancreatic endoderm cells from pluripotent stem cells such as human embryonic stem cells and definitive endoderm cells. The invention is directly applicable to the ultimate generation of pancreatic beta cells that could be used as part of a therapy to treat or even cure diabetes. Additionally, the present invention may be used to generate liver endoderm cells from human embryonic stem cells and definite endoderm cells as well. This invention relates to a method for generating definitive endoderm and pancreatic endoderm cells from stem cells, preferably human embryonic stem cells using defined media in the absence of feeder cells. A simply two step procedure to provide pancreatic endoderm cells from embryonic stem cells represents further embodiments of the present invention.

This disclosure provides an improved system for culturing human pluripotent stem cells. Traditionally, pluripotent stem cells are cultured on a layer of mouse embryonic fibroblast feeder cells to prevent them from differentiating. In the system described here, the role of feeder cells is replaced by defined components added to the culture environment that support rapid proliferation without differentiation. The medium contains an isotonic buffer, a blend of essential nutrients such as protein and lipids, and an effective growth factor or combination of factors that promote proliferation while inhibiting differentiation. Culturing human embryonic stem cells in fresh medium on an extracellular matrix according to this invention causes the cells to expand surprisingly rapidly, while retaining the ability to differentiate into cells representing all three embryonic germ layers. This new culture system allows for bulk proliferation of pPS cells for commercial production of important products for use in drug screening and human therapy.

This invention relates to methods for improved cell-based therapies for retinal degeneration and for differentiating human embryonic stem cells and human embryo-derived into retinal pigment epithelium (RPE) cells and other retinal progenitor cells.

The invention provides a cultivation method of Cyp gene knocked-out rats, and a preparation method of liver microsome of the rats. The Cyp gene knock-out herein includes Cyp single gene knock-out and Cyp multiple gene knock-out. In the method, firstly a Cyp gene knocked-out rat is constructed by means of a CRISPR / Cas system, which includes selection of a knocked-out target site, in-vitro synthesis and transcription of sg RNA and Cas9m RNA, preparation of a pseudopregnant female rat, in-vitro micro-injection and transplanting of a single-cell embryo, and cultivation of the rat, and finally, a homozygote Cyp gene knocked-out rat can be cultured. Furthermore, the liver of the Cyp gene knocked-out rat is extracted and is subjected to homogenization and differential centrifugation to prepare the liver microsome of the rat in Cyp gene deletion. The invention also provides an application of the Cyp gene knocked-out rats and the liver microsome thereof in study on drug metabolism.

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 discloses a complete medium with low serum concentration for cultivating mesenchymal stem cells and a method for cultivating the mesenchymal stem cells using same. The complete medium comprises a cell basic medium, fetal calf serum with final concentration of 1-100 mul / ml, an epidermal growth factor with final concentration of 1-100 ng / ml, and a basic fibroblast growth factor with final concentration of 1-100 ng / ml. The complete medium with low serum concentration successfully reaches equal or even better function of promoting cell proliferation than a culture reagent with high serum concentration. The cultured cells have the typical biological characteristics of mesenchymal stem cells, and can also express an omnipotent mark of the embryonic stem cell and high express the idiosyncratic mark of the neuron under the condition of in vitro inducement. And the difference between the cell batches is little, the cost is low and the security is good. Compared with the prior cultivating method, the method has advantages of simple operation, low probability of pollution and high success ratio of cultivating cells.

Primate embryoid bodies are formed from primate ES cells. The ES cells form clumps. One then removes the clumps, as clumps, and permits incubation under non-adherent conditions. The development of embryoid bodies from primate ES cells is dependent on maintaining the aggregation of cells, as individualized cells will rapidly die.

With respect to a method for differentially inducing embryo-stem cells into hepatocytes, in order to obtain safe hepatocytes that are adequately functionable and able to supply in large quantity, a method for differentially inducing embryo-stem cell into hepatocyte, wherein the embryo-stem cells are cultured in the presence of deletion type hepatocyte growth factor is provided. Further, a method for differentially inducing embryo-stem cells into hepatocytes comprising (a) a step of forming the embryoid body of the embryo-stem cells and (b) a step of culturing the embryoid body in the presence of deletion type hepatocyte growth factor is provided.

The present invention provides a method for inducing differentiation of an embryonic stem cell into a differentiated neural cell. The present invention further provides a method for producing differentiated neural cells, and a population of cells comprising the differentiated neural cells. Additionally, the present invention provides a method for repopulating a spinal cord in a subject, and a method for treating nervous tissue degeneration in a subject in need of treatment. The present invention further provides neural progenitor cells, differentiated neural cells, and uses of same. Also provided is a transgenic non-human animal containing the differentiated neural cells. The present invention is further directed to a method for isolating a population of differentiated neural cells. Finally, the present invention provides a method for identifying an agent for use in treating a condition associated with neuron degeneration.

Disclosed is a method for using a first polar body and a second polar body as well as a single-cell embryo to carry out whole-genome non-exponential amplification and high-throughput genome sequencing, so as to perform preimplantation genetic diagnosis for genetic disease and testing for pathogenic genes causing repeated miscarriages. The method of the present invention comprises the following steps: (1) obtaining oocytes and embryos, and carrying out separation and genome amplification of first and second polar bodies and single-cell embryos; (2) establishing a genome sequencing library and sequencing, and carrying out bioinformatic analysis of the genome to obtain a gene spectrum and information concerning the number of copies of chromosomes and fragments thereof; (3) determining the chromosome ploidy of the polar bodies and embryos as well as information concerning defects, replication and point mutation in the chromosome fragments; (4) selecting normal or suitable embryos for implantation.

A method is provided to prepare a plurality of microwells suitable for the formation of embryoid bodies from embryonic stem cells. The method requires applying an image-forming material to a heat sensitive thermoplastic material in a designed pattern and heating the material under conditions that reduce the size of the receptive material by at least about 60% to create a mold. A polymer such as PDMS is then applied to the mold and removed to form the microwells. In an alternative aspect, the plurality of microwells on receptive material are prepared by etching a microwell designed pattern into a heat sensitive thermoplastic material support and then heating the material under conditions that reduce the size of the receptive material by at least about 60%.

PiggyBac transposons and transposases with enhanced transposition activity in cells are provided. Also provided are associated methods and kits for both introducing exogenous DNA inserts into the genomes of host cells as well as for the removal of the inserts from the host cell genomes. Cells obtained by use of the compositions, methods and kits are also provided.

The invention discloses an inducing method for directionally differentiating human embryonic stem cells to corneal endothelial cells. The method comprises the steps of: cultivating the human embryonic stem cells on a mouse embryonic fibroblast feed layer; sorting human embryonic stem cell clone groups in good state; grafting the groups on a human corneal stromal fibroblast layer processed by mitomycin C and cultivating for 7 days, wherein the human embryonic stem cells are differentiated to rosettes; separating and transferring the rosettes from the human corneal stromal fibroblast layer to a culture bottle; cultivating continuously for 7 days by using a neural crest stem cell culture medium; sorting the neural crest stem cells by a flow cytometry; adding the neural crest stem cells into the culture bottle; placing in a 5% CO2 incubator for incubating and cultivating at 37 DEG C by using a human corneal endothelial cell culture medium; changing the liquid every other day; and cultivating for about 10 days to obtain the corneal endothelial cells. The multiplication capacity of the corneal endothelial cells are similar to that of human corneal endothelial cells and the corneal endothelial cells can be transferred to 1-2 generations in vitro maximally. The corneal endothelial cells can be used as seed cells for cornea construction and transplant in tissue engineering.

The present invention provides a method for producing customized pluripotent stem cells. Specifically, the present invention comprises following steps: extracting proteins from any of the dedifferentiated stem cells or induced pluripotent stem cells, the said dedifferentiated or pluripotent stem cells being prepared by any known method; introducing the protein extract into the adult somatic cells; and culturing the adult somatic cells to produce pluripotent stem cells having the same pluripotency as that of embryonic stem cells. In addition, pluripotent stem cells produced according to the present method and cell therapeutics comprising the same are provided. The method allows pluripotent stem cells to be produced very easily and at a significantly higher yield, compared to typical methods.

This present invention provides novel methods for deriving embryonic stem cells, those cells and cell lines, and the use of the cells for therapeutic and research purposes without the destruction of the embryo. It also relates to novel methods of establishing and storing an autologous stem cell line prior to implantation of an embryo, e.g., in conjunction with reproductive therapies such as IVF.

The invention provides a culture liquid for promoting ectogenesis of embryo vitrified by an open pulled straw (OPS) method after thawing. Frozen 2-cell embryo after thawing is cultured in the culture liquid containing 10<-9> mol / L of melatonin (MT), so that the blastula development rate of the embryo is improved and the number of the cells in the embryo is increased, thereby improving the embryo quality, and providing a beneficial reference for further developing the study of OPS cryopreservation of livestock embryos.

The present invention relates to the generation of neural cells from undifferentiated human embryonic stem cells. In particular it relates to directing the differentiation of human ES cells into neural progenitors and neural cells and the production of functioning neural cells and / or neural cells of a specific type. The invention also includes the use of these cells for the treatment of neurological conditions such as Parkinson's disease.

A method for conversion of an animal into an appropriate recipient of tumor cells derived from a different species. Animals for such purpose can be immuno-incompetent animals that are doubly grafted with orthotopic tissues, in which one grafted tissue (i.e., breast) is from an organ of the same class as the tumor of origin (graft A), and the second grafted tissue (i.e., bone) is from a organ of the same class as a target organ for metastasis (graft B). These dual grafted animals can be used to model human diseases. In one implementation, human tumor cells are orthotopically seeded in graft A in order to analyze the occurrence of metastasis in graft B. Methods and compositions are described for creating a multiorgan human environment in mice, by grafting human stem cells (mesenchymal, embryonic or others) into mice, e.g., injured to enhance specific tissue engraftment. Such chimeric mice can be used to grow human tumors and to study the occurrence of metastasis.

The present invention provides a method of generating megakaryocytes and platelets. In various embodiments, method involves the use of human embryonic stem cell derived hemangioblasts for differentiation into megakaryocytes and platelets under serum and stromal-free condition. In this system, hESCs are directed towards megakaryocytes through embryoid body formation and hemangioblast differentiation. Further provided is a method of treating a subject in need of platelet transfusion.

The invention discloses a micropropagation method of fraxinus rhynchophylla and relates to the micropropagation method. The problems of long reproductive cycle, low reproduction efficiency and poor offspring genetic stability in nursery stock propagation of fraxinus rhynchophylla are solved. The method comprises the following steps of: 1, sterilizing fraxinus rhynchophylla seeds, and culturing single cotyledons of zygotic embryos to obtain cotyledonal cell embryos; 2, performing maturation cultivation on the cotyledonal cell embryos; 3, performing sprouting cultivation on the cotyledonal cellembryos subjected to maturation cultivation to obtain regenerated plants; and 4, transplanting the regenerated plants into a culture medium to culture until new leaves are completely unfolded, and removing a covered plastic thin film. The micropropagation method has the advantages of short nursery stock reproductive cycle, high reproductive rate (culturing for 60 to 70 days) and high reproductionefficiency (each explant can generate 15 to 20 somatic embryos); the germination rate of the somatic embryos is up to 87 to 89.55 percent; the transplanting survival rate of the regenerated plants isup to 75 to 80 percent; and the regeneration plants with the same good characteristics as female parents can be generated in mass.

The present invention relates to a cell culture dish for the erabryoid body formation from embryonic stem cells, which facilitates efficient and stable differentiation of embryonic stem cells by forming embryoid body in grooves on the bottom of a support adhered on the back of the lid for the culture dish by hanging drop culture.

Undifferentiated primordial stem cells are manipulated to permit their long term growth in defined media lacking serum and feeder layer cells by shifting the apoptotic balance of the cells, through increasing the activity of Bcl-2 family anti-apoptotic proteins or decreasing the activity of Bcl-2 family pro-apoptotic proteins. In some embodiments of the invention, the Bcl family protein is Bcl-2. The ES cells sustain the characteristics of undifferentiated, pluripotent stem cells during long-term serum- and feeder layer cell-free conditions, including the ability to be expanded in vitro, but maintain their potential to differentiate into mature cell types.

The present invention relates to a composition for skin regeneration, using a culture medium or a secretion in the culture of an embryonic stem cell-derived endothelial progenitor cell, and to the use thereof. As the present invention uses the secretion in the culture as a medicine and not the embryonic stem cell-derived endothelial progenitor cell itself, the risk of teratoma formation is prevented, and angiogenic activity is promoted to achieve improved effectiveness in healing wounds and burn wounds. The composition of the present invention, when used as a material in cosmetics, increases collagen synthesis and thus prevents skin aging. Particularly, the composition of the present invention in which the secretion in the culture is concentrated into a high concentration provides superior wound-healing effects as compared to a conventional human growth hormone (hGH).

According to the present invention, when nicotinamide is added in a culture process for producing pluripotent stem cells from human differentiated cells, it can increase the efficiency of reprogramming and can significantly reduce the time required for induction of reprogramming. It was verified that nicotinamide inhibits the induction of senescence and oxidative stress in the reprogramming process and increases cell proliferation and mitochondrial activity to effectively improve culture conditions for induction of reprogramming. Particularly, the present invention will contribute to optimizing a process of producing induced pluripotent stem cells from a small amount of patient-specific somatic cells obtained from various sources, and thus it will significantly improve a process of developing clinically applicable personalized stem cell therapy agents and new drugs and will facilitate the practical application of these agents and drugs. In another aspect, according to the present invention, in defined culture conditions in which feeder cells and serum were not used, it was found that nicotinamide can provide a culture medium composition effective for maintaining the undifferentiated state of human embryonic stem cells and human induced pluripotent stem cells, which are typical pluripotent stem cells. The invention can be effectively used for the development of a high-efficiency system for culturing large amounts of human pluripotent stem cells, which is required for the industrialization of human pluripotent stem cells.