40 results about "Mammalian Embryos" patented technology

An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.

A method of generating cells capable of secreting insulin is disclosed. The method comprises subjecting mammalian embryonic stem cells to set of culturing conditions suitable for differentiation of at least a portion thereof into cells displaying at least one characteristic associated with a pancreatic islet cell progenitor phenotype, and subjecting such differentiated cells to a set of culturing conditions suitable for formation of surface bound cell clusters including insulin producing cells.

The invention relates to isolated hemangioblast cells. Hematopoietic and endothelial cells are postulated to be derived from a common progenitor, hemangioblast. While hemangioblast has been isolated retrospectively during embryonic stem cell differentiation, it has not been isolated from embryos or from bone marrow. Prospectively stable clonal cell lines have been isolated from mammalian embryos, from embryonic stem cells and from mammalian bone marrow that can differentiate in vitro into tubular structures with both endothelial and hematopoietic markers such as CD34, CD31, Flk-1, TIE2, P-selectin, Sca-1, thy-1, CD45, and smooth muscle actin. Gene expression profiles in the undifferentiated and differentiated cells were consistent with endothelial and hematopoietic differentiation potential. Transplantation studies in isogenic or immunodeficient mice demonstrated that these cells were not tumorigenic. In an appropriate microenvironment, the cells incorporate into the vasculature and participate in hematopoiesis.

The invention provides methods for the production of transgenic animals comprising a recombinant Ig locus, as well as transgenic antibodies derived therefrom. The methods involve meganuclease cleavage-stimulated homologous recombination in mammalian embryos.

The present invention relates to a method for generating non-human mammalian chimeric embryo. The method involves coculturing denuded non-human mammalian embryos with cells in an Eppendorf micro test tube. The chimeric embryo obtained is then transferred into a non-human recipient mammal so as to develop into a non-human chimeric fetus, non-human chimeric mammal, an embryonic stem cell-derived fetus or an embryonic stem cell-derived mammal.

Mechanisms regulating cell proliferation stop and differentiation initiation during the development stage of mammalian embryo, and the proteins involved therein, are presented. Differentiation regulators, methods of regulating differentiation, transgenic organisms with loss of expression of the differentiation regulator, and methods of preparing the transgenic organisms, are provided.

An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.

Microfluidic embryo scaled channels (14) for handling and positioning embryos provide the opportunity to evaluate and treat embryos in improved manners. Fluid flow is used to move and position embryos within microfluidic channels and channel geometrics may be used to place embryos at specific locations. Surface properties and compliance (deformation) properties of embryos are evaluated as a predictor of viability. The microfluidic channels provide the opportunity for fine controls of pressure to conduct various evalutions at forces slightly below which damage to embryos is known to occur. Measurement of the distance and / or which embryos roll in a same pressure gradient microfluidic channel provides information, with healthy embryos traveling slower or a shorter distance as they demonstrate more stiction to channel walls. Positioned at a constriction (14a, 14b, 24, 26), health embryos also appear to deform less than unhealthy embryos that are more readily pulled into a constriction. In addition, healthy embryos appear to resume their shape better. Fluid from microfluidic channels is easily collected downstream without altering the embryo environment, providing a better opportunity for chemical analysis of fluid chemical analysis than convention manual handling and sampling techniques. Zona pellucida removal of mammalian embryos is achieved as embryos are moved through flow to a precise location where lysing agent can be washed over the embryo to achieve zona removal. Cumulus removal is realized with a series of constrictions to cut cumulus followed by fluid flows to remove cut cumulus from the embryo.

The invention provides methods and systems for assessing the developmental potential of mammalian embryos. The method of the invention comprises taking measurements of cytoplasmic movements in the embryo and / or periodic changes in the shape of the embryo at the single cell stage.

The invention discloses a mammal embryo and oocyte vitrification freezing carrier. The mammal embryo and oocyte vitrification freezing carrier comprises a freezing tubule and a tapered sucking head both of which are mounted in a matching and butting manner, wherein a metal weight body is arranged at the back end of the freezing tubule; and a sealing interlayer is arranged between the metal weightbody and an inner storage cavity of the freezing tubule. The mammal embryo and oocyte vitrification freezing carrier is low in cost as the manufacturing materials are the common materials, is easy tooperate and is capable of controlling amount of a freezing liquid precisely and freezing multiple oocytes and embryos for once. As the metal weight body is adopted, a sample cannot float on a liquid nitrogen surface and the sample cannot be lost; freezing damage of the sample is reduced due to the fast freezing speed and high freezing efficiency; and the carrier can be utilized repeatedly and cannot break to cause lost of the sample, so that cost is saved. Marks can be made on the tubule, so that convenience can be brought about for long-term preservation.

The invention discloses an in vitro micro-dynamic culture device of mammalian embryos and a method for cultivating embryos by using the same, relates to an in vitro culture device of embryos and a cultivation method using the same, and overcomes the defects of an in vitro oscillating micro-dynamic incubator of mammalian embryos and the defects of an existing patent technology. The culture device comprises a glass micro-fluid channel layer, a main body layer, an air chamber seal cover, an air duct, an injector, an electric motor and a square wave generator. The cultivation method comprises the following steps: 1, setting the frequency of the square wave generator and the stretching speed of the electric motor; 2, injecting a culture solution and mineral oil, and putting the embryos into a culture room; and 3, powering on a switch, and cultivating. The in vitro micro-dynamic culture device is applied to the field of biology.

The present invention relates to complement proteins, in particular, C3 protein, and methods using the same for enhancing the development of preimplantation mammalian embryos in vitro for use in assisted reproductive technologies. In particular, the present invention relates to supplementation of complement C3 protein, its precursors, fragments, or derivatives, to culture media to improve the development of cultured embryos and, thereby, enhance pregnancy rates of in vitro fertilization.

The invention relates to a method for preparing and separating stereotyped endoderm cells by using monolayer culture technology and also discloses culture mediums suitable for preparing the stereotyped endoder mcells. The culture mediums comprise Neurobasal culture medium, DMEM / F12 culture medium, N-2 additive, B-27 additive, bovine serum albumin, mono-thioglycerin and activin A. The inventive method is simple and efficient, and can improve the efficiency in differentiating stereotyped endoderm of embryonic stem cells (preferably inhuman mammals) greatly.

A method of enhancing in vitro development of a mammalian embryo is disclosed which comprises supplementing the culture medium with a prostaglandin, or a prostaglandin analog, in an amount effective to promote complete hatching of the embryo (i.e., freeing of the embryo from the zona pellucida). The quality of human blastocysts is enhanced in vitro by culturing with a prostacyclin agonist, Iloprost. The in vivo implantation potential and live birth potential of an in vitro fertilization embryo is thereby enhanced and establishment of a viable pregnancy is facilitated.

Provided are a base editing composition comprising deaminase and target-specific nuclease, a base editing method using the base editing composition, and a method for producing a genetically modified animal. The base editing composition has a base editing activity in mammal embryos.

The invention relates to a PCR augmentation primer and agent box and the application in the sex identification of mammals, which is a pair of primer augmenting the 163bp area in gene Sry, sequence of basic group is: SRY5: 5'-TGAACGCCTTCATTGTGTGGTC-3',SRY3: 5'-GCTAGTAGTCTCTGTGCCTCCT-3'. The main components of the agent box are: buffer for DNA extraction, Taq DNA polymerase, PCR reaction liquid, blank control, paraffin oil and sample buffer. The applying steps of agent box in the sex identification of mammals are: (1) sample treatment, (2) augmentation, (3) product detecting. The invention is characterized by judging the existence of Y colorant layer specific fragment by detecting the augmenting product, and the avoidance of the unnecessary pollution, and the suitness for the quick(about 1 hour) sex identifying of mammal embryo in practice, the accuracy is 100%.

The present invention provides a method of isolating a pluripotent cell from a pre-implantation embryo without isolation of the pluripotent cells from other cells, the method including propagating a whole pre-implantation embryo including one or more pluripotent cells, embedded in a feeder cell layer and cultivated in a medium substantially free of serum, and isolating a pluripotent cell from the one or more pluripotent cells. The present invention also provides pluripotent cells generated by the method and uses thereof.

The invention relates to isolated hemangioblast cells. Hematopoietic and endothelial cells are postulated to be derived from a common progenitor, hemangioblast. While hemangioblast has been isolated retrospectively during embryonic stem cell differentiation, it has not been isolated from embryos or from bone marrow. Prospectively stable clonal cell lines have been isolated from mammalian embryos, from embryonic stem cells and from mammalian bone marrow that can differentiate in vitro into tubular structures with both endothelial and hematopoietic markers such as CD34, CD31, Flk-1, TIE2, P-selectin, Sca-1, thy-1, CD45, and smooth muscle actin. Gene expression profiles in the undifferentiated and differentiated cells were consistent with endothelial and hematopoietic differentiation potential. Transplantation studies in isogenic or immunodeficient mice demonstrated that these cells were not tumorigenic. In an appropriate microenvironment, the cells incorporate into the vasculature and participate in hematopoiesis.

The disclosure relates to a method of introducing mRNA encoding Cas9 protein (Cas9 mRNA) into a mammalian embryo, comprising the steps of;(a) placing a mixture of the mammalian embryo and a solution comprising Cas9 mRNA in the gap between a pair of electrodes, and(b) applying a voltage to the electrodes for a voltage application duration, wherein the voltage and the voltage application duration achieve the efficiency of mRNA introduction (R) higher than the minimum required efficiency of mRNA introduction (Rmin) that is calculated on the basis of the concentration of Cas9 mRNA (ng / μl).

The invention relates to the technical field of sex determination, in particular to a mammalian embryo sex determination method. According to the method, by inserting a fluorescent reporter gene intoa Y chromosome, and by means of fluorescent expression of the fluorescent reporter gene, the embryo sex of a mammal is determined accurately, and the method is simple, efficient, accurate and small indamage; in the process of building a male mammal modal with the Y chromosome containing the fluorescent reporter gene, spacer regions located at Ddx3y and Uty genes are selected, exogenous gene integration is conducted for a target area, and the influence of target site selection on a transgenic animal is lowered; in the gene editing process, by adopting Cas9 protein and single guiding sgRNA designed according to a target area gene sequence, target sites DNA can be cut accurately, the fluorescent reporter gene is inserted between the Ddx3y and Uty genes of the Y chromosome directionally, andthe gene knock-in means is efficient and simple.

The invention provides methods and systems for assessing the developmental potential of mammalian embryos. The method of the invention comprises taking measurements of cytoplasmic movements in the embryo and / or periodic changes in the shape of the embryo at the single cell stage.

Provided are a base editing composition comprising deaminase and target-specific nuclease, a base editing method using the base editing composition, and a method for producing a genetically modified animal. The base editing composition has a base editing activity in mammalian embryos.

The invention relates to a sex identification method for a mammalian embryo. The sex identification method for the mammal embryo comprises the following steps: (1) obtaining the embryo; (2) obtaining genome DNA of embryonic cells; (3) designing and synthesizing primers and probes; (4) performing QPCR reaction: during PCR amplification, adding a pair of primers and a specific fluorescent probe at the same time; and (5) amplifying the curve and analyzing, and identifying the sex of the embryo. The sex identification method for the mammal embryo, disclosed by the invention, is a real-time fluorescent quantitative PCR (Quantitative Real-time PCR) probe method, is simple in design, only needs a pair of primers and is added with a specific fluorescent probe at the same time, so that the limitation that only end-point detection can be carried out in a traditional method is effectively solved; the method has the advantages of less sampling, high sensitivity, stronger specificity, rapidness, simplicity, convenience, effective solving of a PCR pollution problem, good universality, high automation degree and the like.

The present invention describes novel compositions for deriving, maintaining and growing pluripotent and germ-line competent mammalian embryonic stem cells. The compositions of this invention refer to compositions comprising a 1) conditioned medium of a cell line expressing limited amounts of Leukemia Inhibitory Factor (LIF), 2) conditioned medium from a cell line transfected with mammalian LIF and 3) a medium supplemented with recombinant rabbit LIF. The present invention describes novel compositions for deriving, maintaining and growing adult human stem cells and / or adult early progenitor cells, preferably under stroma-free conditions and without added LIF and / or cytokines or growth factors. The media of the present invention are used for the generation of pluripotent and germ-line competent embryonic stem cells of mammals of which these cells were not obtained up to now. The media of the present invention are used for the generation of adult human stem cells and / or adult early progenitor cells. The present invention is also directed to a novel rabbit LIF and to nucleotides encoding the rabbit LIF and methods for the expression of recombinant rabbit LIF in the Pichia pastoris expression system.

The invention relates to a method for identifying the sex of a mammal embryo. The method for identifying the sex of the mammal embryo comprises the following steps: (1) obtaining the embryo; (2) obtaining genome DNA of the embryonic cells; (3) designing and synthesizing primers; (4) performing QPCR reaction: adding a fluorescent dye into a PCR reaction system; and (5) amplifying the curve and analyzing, and identifying the sex of the embryo. The method for identifying the sex of the mammal embryo, disclosed by the invention, adopts the real-time fluorescent quantitative PCR method, is simple in design, only needs a pair of primers, can analyze and detect the specificity of amplification reaction through a melting point curve, and has the advantages of high sensitivity, short time consumption, low initial cost, good universality, high automation degree and the like.

Described herein is a method for treating or restoring loss of hearing and balance in a subject in need thereof. In certain aspects, the disclosed method is comprised of transplanting a humanized otocyst to a target delivery site in the subject. According to certain aspects, the otocyst is obtained from a humanized non-human mammal embryo. In certain exemplary aspects, the non-human mammal is a pig. In certain embodiments, prior to transplantation, the otocyst is cultured in a species-specific tissue culture buffer.

The invention discloses an in vitro micro-dynamic culture device of mammalian embryos and a method for cultivating embryos by using the same, relates to an in vitro culture device of embryos and a cultivation method using the same, and overcomes the defects of an in vitro oscillating micro-dynamic incubator of mammalian embryos and the defects of an existing patent technology. The culture device comprises a glass micro-fluid channel layer, a main body layer, an air chamber seal cover, an air duct, an injector, an electric motor and a square wave generator. The cultivation method comprises the following steps: 1, setting the frequency of the square wave generator and the stretching speed of the electric motor; 2, injecting a culture solution and mineral oil, and putting the embryos into a culture room; and 3, powering on a switch, and cultivating. The in vitro micro-dynamic culture device is applied to the field of biology.

An object of the present invention is to provide a means for efficiently obtaining mammalian embryos having high conception rates. A first aspect of the present invention is a method for selecting a mammalian embryo prepared by in vitro culture from a fertilized egg, comprising a step of selecting an embryo using two or more of the following indicators: the time from fertilization to the completion of first cleavage; the morphology at a stage after first cleavage and before second cleavage; the morphology at a stage after third cleavage and before fourth cleavage; and the amount of oxygen consumed at the early blastocyst stage, the blastocyst stage, or the expanded blastocyst stage.

The invention discloses a culture method for improving the survival rate of mammalian embryos in vitro, which comprises the following steps: (1) taking sheep oocytes from ewes and putting them into a mature culture solution, culturing them in an incubator for 20-24 hours, Obtain mature sheep oocytes; (2) Put the mature sheep oocytes and sperm into the fertilization medium, culture them in the incubator for 18‑24 hours, and fertilize them to obtain early sheep embryos; (3) Put the The fertilized sheep embryos were washed repeatedly with TCM199 culture medium, then put into sheep embryo in vitro culture medium, and placed in an incubator for in vitro culture of embryos. The temperature in the incubator was maintained at 38-39°C, and the gas concentration was maintained at 5% CO2; wherein, the above-mentioned incubator includes a cultivation chamber, a chamber door, and a gas concentration adjustment chamber connected with the cultivation chamber; there are multiple cultivation chambers in the cultivation chamber, and a culture medium is respectively placed in the cultivation chambers racks and partitions. The stability of the environment in the incubator of the present invention ensures the survival rate of embryos.