238 results about "Cells embryo" patented technology

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.

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 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.

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 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).