183 results about "Blastocytes" patented technology

Methods are disclosed for generating HLA homozygous parthenogenetic human stem cell (hpSC-Hhom) lines from both HLA homozygous and HLA heterozygous donors. These hpSC-Hhom lines demonstrate typical human embryonic stem cell morphology, expressing appropriate stem cell markers and possessing high levels of alkaline phosphatase and telomerase activity. Additionally, injection of these cell lines into immunodeficient animals leads to teratoma formation. Furthermore, in the case of HLA heterozygous donors, the hpSC-Hhom lines inherit the haplotype from only one of the donor's parents. SNP data analysis suggests that hpSC-Hhom lines derived from HLA heterozygous oocyte donors are homozygous throughout the genome as assessed by single-nucleotide polymorphism (SNP) analysis. The protocol as disclosed minimizes the use of animal-derived components, which makes the stem cells more practical for clinical application.

A method for isolating an inner cell mass comprising the steps of immobilizing a blastocyst stage embryo having a zona pellucida, trophectoderm, and inner cell mass, creating an aperture in the blastocyst stage embryo by laser ablation, and removing the inner cell mass from the blastocyst stage embryo through the aperture. The aperture is through the zona pellucida and the trophectoderm. The laser ablation is acheived using a non-contact diode laser. The inner cell mass removed from the blastocyst stage embryo is used to establish human Embryonic Stem Cell lines.

Instead of immersing human reproductive cells in a single culture medium throughout the various procedures used in IVF, a process is provided by which the reproductive cells may be moved through a sequence of distinct culture media as the various IVF procedures are carried out. In one implementation, the culture media specifically formulated to provide a physical environment similar to that found within the female reproductive tract and conducive to growth and development of human reproductive cells during the various stages of the IVF process. In this regard, specifically formulated culture media can be applied to support the reproductive cells in one or more of the following procedures: oocyte retrieval and handling; oocyte maturation; ordinary fertilization; oocyte, zygote and embryo examination and biopsy; embryonic development to the eight-cell stage; embryonic development to the blastocyst stage; embryo transfer; and cryopreservation.

The invention discloses a split-range embryo culture solution and a preparation method thereof, belonging to the technical field of cytology and biology. The split-range culture solution added with three embryotrophic factors comprises a cleavage solution and a blastula solution, wherein the cleavage solution and the blastula are respectively used in a fertilized egg before and after 8-cell cycle. The embryotrophic factors added into the embryo culture solution disclosed by the invention have synergistic effect, so that the defect that the split-range culture solution loses the embryotrophic factors secreted by an embryo is avoided, the fertilized egg has higher embryo stem cell pluripotency during the growth in the culture solution disclosed by the invention, the development situation of the fertilized egg is equivalent to the in-vivo embryo growth, the in-vitro growth and development of the fertilized egg is improved, and the quality of the embryo is improved.

A novel population of multipotent cardiac precursor (MCP) cells derived from human blastocysts derived stem cells is disclosed, methods for the preparation thereof and use of the cells for in vitro testing. Basement cells derived from hBS cells are also disclosed and method for the preparation of MCP cells from basement cells. The MCP cells have the following characteristics

• • i) at least 1% of the cells exhibit no antigen expression of one or more markers for undifferentiated cell, the marker being selected from the group consisting of SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and Oct-4,
  • ii) at least 1% of the cells exhibit no protein expression of one or more of a neural marker including nestin or GFAP
  • iii) at least 1% of the cells exhibit protein and/or gene expression of one or more of a mesodermal marker including brachyury, vimentin or desmin
  • iv) at least 1% of the cells exhibit protein and/or gene expression of Flk-1 (KDR).

Furthermore, the MCP cells have a characteristic morphology. They grow as clusters of small, round and phase-bright cells; individual cells are 5-20 μm in diameter and each cluster is composed of 2-500 cells. They form clusters of round or elongated shape, that appear as loosely adherent cell clumps that as illustrated in FIG. 2 panel a, b and c. Furthermore, they have a relatively high nucleus-to-cytoplasma ratio, e.g. 1:2-1:64 of the total volume of the cell and/or appear as balloons on a string, as illustrated in FIG. 18, schematic sketch. Moreover, the MCP cells are non-contracting.

The invention discloses a swine in-vitro embryo culture solution. The swine in-vitro embryo culture solution disclosed by the invention is obtained through adding ligustrazine, with the final concentration of 0.05-5.00 microgram/milliliter, into an embryo culture solution NCSU-23. A traditional Chinese medicine monomer component, namely ligustrazine, is added into the swine in-vitro embryo culture solution disclosed by the invention; shown by experiments, through carrying out in-vitro culture on the swine parthenogenetic activated embryos and the swine in-vitro fertilized embryos by using the culture solution, the development efficiency of the embryos and the number of the cell masses and total cells in blastulae can be effectively increased. The swine in-vitro embryo culture solution has the advantages that a foundation is laid for the development of other biotechnologies, medical research and animal husbandry, and meanwhile, a certain theoretical basis for clarifying the development mechanism of early swine embryos is provided.

The invention provides a method for improving the blastocyst rate of in-vitro embryos of animals. According to the method, the developmental capacity of a parthenogenetic embryo and an in-vitro fertilization embryo can be remarkably improved by adding type I interferon into an embryonic development culture solution for the in-vitro culture of the embryos; compared with the blastocyst rate obtained under the culture of the conventional culture solution, the blastocyst rate is improved by 7 to 13 percent; and the blastocyst hatching rate is also remarkably higher than the blastocyst hatching rate under the culture of the conventional culture solution. By the method, the early development capacity of in-vitro embryos of cattle and sheep is improved, so that the in-vitro production efficiency of embryos of the cattle and sheep is improved, and high-quality embryos are provided for the scientific research and production field in which the embryos of the cattle and sheep are taken as materials.

The invention discloses multifunctional vulvae care solution and a preparation method thereof. The care solution is prepared by mixing the following Chinese traditional medicines by weight portion to obtain the medicine solution by decoction and then adding with the following western medicines by weight portion; the Chinese traditional medicines are as follows: 3 potions to 7 portions of ebony, 4 portions to 8 potions of cuttlebone, 1 portions to 3 portions of white vitriol, 15 portions to 25 portions of herba houttuyniae, 12 portions to 18 portions of radix isatidis, 12 portions to 18 portions of Chinese pulsaiila root, 7 portions to 13 portions of glabrous greenbrier rhizome, 7 portions to 13 portions of radix sophorae tonkinensis, 7 portions to 13 portions of belamcanda bhinensis DC., 15 portions to 25 portions of dayflower, 15 portions to 25 portions of groundsel, 12 portions to 18 portions of agrimony, 7 portions to 13 portions of raspberry and 3 portions to 7 portions of Chinese tumion seed; the added western medicines are vitamin B2 metronidazole, jasmine top note flavors, benzalkonium bromide solution and jie-er-yin liquid; stilbestrol and/or philter and/or saxol. The pH value of the care solution is about 6.5, and the care solution can inhibit the reproductions of bacteria and blastocyte, adopts a plurality of heat-clearing and detoxifying drug, and has the functions of sterilization, desinsection, relieving itching and anticancer as well as detoxification of cervical erosion.

The invention provides a kind of transgenic inhuman mammal by producing klone gene without mouse model. Roll outside self murder gene into the mammal. The expression box contains promoter, self murder gene relation with promoter and termination codon. The natural masculine gene will express in testes organize after transgenic animal builds system. The transgenic animal can provide blastosphere for eliminating gene technic to improve the efficiency of the technic and to solve the problem that mouse genetic background is impure.

The invention relates to the use of cells from cell lines, which are derived from early embryonic stages, for the donor-specific increase in transplantation tolerance and for repairing damaged tissue. Areas of application of the invention include the field of medicine and the pharmaceutical industry. The aim of the invention is to produce a donor-specific immunotolerance in order to prevent a rejection of the transplanted tissue due to an immune response and thus to be able to limit the administration of immune suppressive agents. In order to produce a donor-specific immunotolerance, embryonic stem cell-like cell lines (ECL) are obtained from blastocysts and are transfected with genetic material of the donor, which codes for the MHC haplotypes. The cells produced in such a manner are administered to the recipient before the transplantation for producing an immunotolerance against the tissue to be transplanted or for regenerating already damaged tissue.

The invention discloses a kit for screening high-quality blastulas. According to the kit, birth safety is forecasted before a human assisted reproduction blastula is implanted, and the kit comprises products of expression quantities of 398 genes as shown in a table 2 in wall trophoblast cells and polar trophoblast cells. The human assisted reproduction blastula is normally developed, and a pregnant woman normally delivers if the expression quantity of each gene in the 398 genes of the wall trophoblast cells of the human assisted reproduction blastula (a blastula implanted before fifth days of in-vitro fertilization) is twice or more or half or less that of the same gene in the 398 genes of the polar trophoblast cells. The kit has important application values for scientific judgment of transplantation strategies and decisions and possibility reduction of perinatal period complications of pregnant women and sick children in assisted reproduction.

The invention discloses a kit for predicting birth safety before people-assisted reproduction blastosphere implantation. The kit comprises products, wherein the products are used for detecting the expression quantity of 97 genes shown in the table 5 and from a cell of a trophectoderm of a people-assisted reproduction blastosphere. If the expression quantity of the 97 genes from the cell of the trophectoderm of a people-assisted reproduction blastosphere meets the evaluation standard, the people-assisted reproduction blastosphere can develop normally and a pregnant woman can have eutocia; otherwise, the people-assisted reproduction blastosphere cannot develop normally, the possibility of affecting baby health and occurring perinatal complication is increased along with the increase of the number of the genes not meeting the evaluation standard, and the transplantation strategy and decision-making are further affected. With adoption of the kit, the possibility of reducing the perinatal complication and the number of babies suffering from illnesses is reduced.

The invention discloses a method for preparing an expression vector of adenoviruses. The method comprises the following steps: using a genome of wild type adenoviruses AdC6 as a base, deleting an E1 coding region and an E3 coding region according to a genome direct cloning method, and inserting I-Ceu I and PI-Sce I digestion sites in the E1 deleted region. Simultaneously, OFR6 of human-derived adenoviruses Ad5 is used for replacing ORF6 of AdC6, the success rate of packaging recombinant viruses and the titer of the packaged recombinant viruses in human-derived kidney blastocyte (293A) are improved, and the replication-deficient expression vector of the recombinant adenovirus is obtained. According to the direct molecular cloning method, long-fragment plasmids are constructed, the method issimple and easy to operate, and the method can be used for construction of any long-fragment vectors. The construction of the expression vector of the adenoviruses can be completed through several times of cloning connection, and the steps are simple; the E1 and E3 regions are deleted, the Ad5 ORF6 is replaced, and the deletion and replacing can be completed in the PCR amplification process of DNA fragments; the method is easier and quicker in horizontal bacteria screening and positive cloning, and the construction time of the adenovirus vector is shortened.

The invention discloses a special culture medium and method for culturing porcine trophoderm stem cells. The special culture medium is composed of bFGF, activin A, Y27632, Knockout SR and beta-mercaptoethanol. The culture method comprises the following steps: previously coating a stem cell culture dish with an extracellular matrix; digesting porcine blastula with proteinase to remove the zona pellucida, transferring into the stem cell culture dish, and adding the special culture medium; and culturing until a trophoderm stem cell cloning group with the total cell count of 1000-2000 is formed, digesting into unicells, transferring into a new extracellular-matrix-coated culture dish, and carrying out cell subculture. The special culture medium has the advantages of definite components and high use safety, and avoids the pollution of heterogenous cells. The culture method is simple and efficient, and has the advantages of high cloning formation rate, low apoptosis rate, high proliferation and subculture capacity, high safety and high stability when being used for porcine trophoderm stem cell culture.

The invention provides a constructing method of 1.3 copy C gene type HBV transgenic mice. Through blastocyst injection of ES cells, the method prepares targeted integration 1.3 copy C gene type HBV (Hepatitis B virus) transgenic mice. First, targeted integration1.3 copy C type HBV expression vectors are constructed, and then an electrical transduction transformation method is employed to transform recombinant plasmids into ES cells; after the and cultivation, targeted integration recombinant ES cells are obtained; then the targeted integration recombinant ES cells are digested into single cells and injected into blastocysts; and then the blastocysts are transplanted to uteruses of pseudopregnant female mice. The method sets up the 1.3 copy C type HBV transgenic mice for the first time home and abroad, is specifically applied to the field of hepatitis B related researches, and provides a more ideal animal model for the researches on prevention and treatment of hepatitis B in China.

The invention relates to the technical field of medical artificial intelligence, in particular to an embryo development potential prediction method and system, equipment and a storage medium. The method comprises the steps: inputting an embryo initial image into a blastocyst prediction model, and obtaining an embryo feature vector; inputting the embryo feature vector into a bidirectional long-short-term memory network to obtain embryo development features; based on a cross-modal feature fusion mechanism, obtaining fusion features according to clinical data and the embryonic development features; and inputting the fusion features into a first multi-layer sensor, and predicting to obtain the embryo pregnancy rate. According to the invention, multi-focal-segment embryo videos shot in the early stage are analyzed, and the fusion features with the space-time characteristic is obtained by utilizing a multi-focal-segment selection model and a time transfer model, so that the pregnancy rate of the embryo cultured in vitro is predicted in real time, and the prediction accuracy is improved; meanwhile, by predicting the blastocyst forming probability and the euploid probability, doctors are assisted in early embryo screening, and therefore the labor cost is reduced.

The invention discloses a breeding method for transgenic pigs expressing sIFITM3 (swine interferon induced transmembrane 3) genes. The breeding method comprises the steps as follows: firstly, the construction of swine fibroblast cell lines stably expressing the sIFITM3 genes comprises the following steps: the step a refers to the construction and the linearization of pcDNACAsIITM3 vectors, wherein, primers are designed, lymph node tissue RNA (Ribose Nucleic Acid) of pigs is extracted, the sIFITM3 genes are obtained through RT-PCR (Reverse Transcription-Polymerase Chain Reaction) amplification, ethanol precipitation is carried out, and sterile water is used for dissolving; the step b refers to liposome transfection, wherein, swine fibroblasts expressing sIFITM3 are constructed; the step c refers to screening and proliferation of nuclear donor cells transfected with the sIFITM3 genes; secondly, recombination blastula acquisition based on a manual nucleus transplantation method comprises the following steps: the step a refers to the preparation of recipient cells; the step b refers to enucleation and injection of the nuclear donor cells; and thirdly, the embryo transplantation and the acquisition of the transgenic pigs comprise the following steps: the step a refers to embryo transplantation, wherein, bred blastulas are transferred to a fallopian tube of a surrogate sow, and then detection is carried out; and the step b refers to transgenic individual identification. The breeding method is feasible, and is simple and convenient to operate; in addition, the pigs are the transgenic pigs expressing sIFITM3, and have the potential capability to resist animal virus such as foot-and-mouth disease virus, Japanese encephalitis virus, swine influenza virus and the like.

The invention discloses a method for culturing a single porcine embryo in vitro, and belongs to the field of agriculture-animal husbandry and veterinary medicine. The method particularly involves thatfeeder layer cell micro-droplets are adopted to be co-cultured with a single embryo so as to obtain a better development result; the development process of single embryo culture is consistent with the development process of traditionally used multi-embryo culture, and the single embryo attached with a transparent belt or removing a transparent belt can obtain a porcine parthenogenetic developmentblastosphere in 7 days. The method for culturing the single porcine embryo in vitro has the advantages that the provided feeder layer cell micro-droplets preparation is simple, a reliable original material for studying the development of a single embryo can be provided according to the provided method, and the needs of tracking, observing and further researching the development process of the single porcine embryo can be fully met; the provided single embryo culture method can be widely applied to single embryo culture of various species.

The present invention provides compositions and methods for regulating fertility in mammals. In general, the invention relates to a novel protein produced by oocytes named JY-1, and nucleic acids encoding the JY-1 protein, for controlling folliculogenesis and early embryonic development, particularly in monoovulatory species. In particular, the present invention provides nucleic acid and amino acid sequences encoding JY-1, vectors for the expression of JY-1, host cells expressing JY-1, RNAi probes for reducing levels of JY-1 message, and antibodies to JY-1. Specifically, developing and mature oocytes express JY-1 in vivo, while granulosa cells treated in vitro with recombinant JY-1 (rJY-1) protein reduced cell proliferation while increasing progesterone synthesis and estradiol production. Further, reducing JY-1 protein in developing embryos in vitro using inhibitory siRNA constructs corresponded with arrested blastocyte maturation.