309 results about "Blastocyst" 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 of vitrifying mammalian cells. According to the method of the present invention, biological cells of mammalian origin are frozen quickly by a vitrification method. Upon exposure to a coolant, the biological cells undergo vitrification. The biological cells which have undergone vitrification may be stored for a period of time and then devitrified at a later date. The devitrified biological cells remain viable. Preferred biological cells according to the present invention are developmental cells including blastocysts, embryos, and oocytes.

A stem cell library is created by genetically modifying stem cells with nucleic acids encoding polypeptides which can promote stem cell differentiation into specific cell types. Alternatively, the stem cell library is exposed to an externally added factor that promotes stem cell differentiation into a desired cell line, e.g., neuronal or muscle. The library is used to determine the effect of the encoded protein on the differentiation process. The library is also used to produce nucleic acids for insertion into embryonic stem cells to produce transfected embryonic stem cells. The nucleic acids are inserted into a locus that permits widespread expression of the encoded polypeptide in animals produced from blastocysts that incorporate the transfected cells. Non-human chimeric animals produced by combining blastocysts derived from animal models of human disease and embryonic stem cells transfected with molecules from the library provide an in vivo system for therapeutic design.

An automated system and method of cryopreservation and reanimation of oocytes, embryos, or blastocysts is disclosed. One or more oocytes or embryos are positioned in a processing container, the processing container being configured to allow fluid to flow into and out of the processing container, where two or more fluids flow into and out of the processing container with oocytes or embryos therein. The temperature of the fluid may be controlled in the processing container according to predetermined requirements. The flowing of the fluids may be controlled by a central controller adapted to control one or more valves.

The invention discloses a human ovocyte cryoprotectant. The human ovocyte cryoprotectant contains an ovum refrigerating fluid and an ovum thawing fluid, wherein the ovum refrigerating fluid contains a balance fluid and a refrigerating fluid; the ovum thawing fluid contains a resuscitation fluid, a diluent-1, a diluent-2 and a cleaning solution. The human ovocyte cryoprotectant is developed by combined application of the propanediol and glycol permeable cryoprotectants, and taking the trehalose (trehalose, alpha-D-glucopyranosyl alpha-D-glucopyranoside, C12H22O11) as a non-permeating agent. The non-permeating cryoprotectant suitable for freezing the human ovocyte is developed so that the formation of the ice crystal in the cell can be inhibited, the function of the frozen ovum can be protected and the developmental potentiality of the frozen ovocyte can be greatly improved. The clinical result proves that the fertility rate, the cleavage rate and the high quality blastocyst rate of the frozen ovocyte can be greatly improved.

An in vitro culture and / or fertilization medium containing N-acetylcysteine amide (NAC amide) reduces or prevents oxidative stress and free radical formation that contribute to the cellular damage and eventual demise of sperm, oocytes and embryos that are cultured, fertilized and maintained in vitro. The NAC amide-containing medium composition for in vitro culture and fertilization is suitable for use in the culture of oocytes, in the culture and development of early embryos, in the preparation or culture of sperm, and in the pre-treatment of oocytes or sperm. The NAC amide-containing composition supports the growth of viable embryos until blastocyst stage.

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.

Methods, compositions and kits for determining the likelihood of reaching the blastocyst stage for one or more embryos or pluripotent cells are provided. These methods, compositions and kits find use in identifying embryos and oocytes in vitro that are most useful in treating infertility in humans.

Methods, compositions and kits for determining the developmental potential of one or more embryos are provided. These methods, compositions and kits find use in identifying embryos in vitro that are most useful in treating infertility in humans.

The present invention represents an improvement upon the basic method of generating genetically modified animals, particularly knock-outs, through the injection or aggregation of genetically modified foreign cells with wild type blastocysts or embryos. This improvement comprises the use of animals with permanent or conditional fertility defects as the source of blastocysts / embryos in this process. By using such animals, germ cells derived from the genetically modified foreign cells have a competitive advantage in the process and the frequency of producing genetically modified animals is increased. Chimeric blastocysts or aggregates produced according to this improved method are also part of the invention. In another aspect, the invention teaches the generation and use of an inducibly sterile animal. A method for rendering wild type blastocysts or early stage embryos sterile for reproduction is also contemplated as part of the invention.

An improved method for vitrification of biological cells, especially blastocyst-derived stem cells (BS cells). The method is very mild for the cells that remain viable after they have been thawed. The method comprises, i) transfer of the cells to a first solution (solution A), ii) optionally incubation of the cells in the first solution, iii) transfer the cells obtained in step i) or ii) to a second solution (solution B), iv) optionally incubation of the cells in the second solution, v) transfer of the cells obtained from step iii) or iv) into one or more closed straws with dimensions that allow a volume of at least 20 μl to be contained in them vi) sealing the one or more closed straws, and vii) vitrification of the one or more closed straws. An important feature of the present invention is the use of closed straw and that relatively large volumes can be efficiently vitrified and subsequently thawed.

The present invention relates to the use of an oxytocin receptor antagonist in females undergoing embryo transfer as part of an assisted reproductive technology. In particular, methods are provided for increasing ongoing implantation rate, increasing ongoing pregnancy rate, increasing clinical pregnancy rate, and / or increasing live birth rate in a female subject undergoing embryo transfer. Specifically, the antagonists are released in the luteal phase when the endometrium is receptive for embryo implantation and / or when the embryo has reached the blastocyst-stage.

The invention discloses an in-vitro maturating method for sheep oocyte, a pretreatment solution and a kit. The related pretreatment solution applied to in-vitro maturating of sheep oocyte is characterized by comprising sheep small follicle fluid and routine culture-solution compositions for in-vitro maturating of sheep oocyte. The disclosed in-vitro maturating method for sheep oocyte is characterized in that a pretreatment step is increased on the basis of a conventional in-vitro maturating method. Also, experiments in the invention verify that oosperm obtained after an oocyte which is cultured by employing the maturating method is subjected to in-vitro fertilization has the cleavage rate and the blastocyst rate both substantially higher than those of an oocyte cultured by employing a conventional maturating method. The provided pretreatment solution and the in-vitro maturating culturing method are capable of effectively promoting in-vitro maturation of sheep oocyte, and have the characteristics of no hazard to occyte, few limits, good effect and the like.

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 characteristicsi) 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 GFAPiii) at least 1% of the cells exhibit protein and / or gene expression of one or more of a mesodermal marker including brachyury, vimentin or desminiv) 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.

A method for the transfer of human blastocyst-derived stem cells (hBS cells) to feeder-free culture system and propagation of the cells in such a feeder-free culture system, the method comprising the following steps of (a) transferring the balstocyst-derived stem cells from feeder to feeder free culture by mechanical treatment, (b) optionally, culturing the blastocyst-derived stem cells under feeder cell free growth conditions in a suitable growth medium and / or on a suitable support substrate, and (c) optionally passaging the blastocyst derived stem cell line every 3-10 days by enzymatic and / or mechanical treatment. The invention also relates to the application of hBS cells cultured under feeder free condition in medicine (e.g., myocardial regeneration) and screening and toxicity tests.

The invention relates to canine embryonic stem cells, methods of cultivation and propagation of the cells, and production of differentiated cells. The embryonic stem cells may be obtained by isolating a canine embryo, culturing the embryo in the presence of a feeder layer and one or more proliferation agent, removing a blastocyst outgrowth, and culturing the outgrowth in the presence of a fresh feeder layer.

The invention discloses a blastocyst nutrient solution and a preparation method thereof. The blastocyst nutrient solution comprises a composition for treating and culturing a zygote, a bactericide, a selectively added indicator and a selectively added human serum albumin. Each index of the blastocyst nutrient solution reaches the standard of an in-vitro nutrient solution, the use is safe, and whether the nutrient solution is deteriorated or not can be easily determined, and therefore a powerful guarantee is provided to the assisted reproductive technology.

The invention relates to a method for bovine in-vitro fertilized embryo culture and a used culture solution. Particularly, on one hand, the invention relates to a method for culturing a bovine in-vitro fertilized embryo. The method comprises the following steps of putting the bovine in-vitro fertilized embryo into a bovine in-vitro fertilized embryo culture solution and carrying out embryo in-vitro culture under the conditions that the temperature is 38.5 DEG C, the content of CO2 is 0.5% and the humidity is 100%. The invention further relates to a bovine in-vitro fertilized embryo culture solution. The culture solution comprises NaCl, KCl, NaHCO3, MgCl2, KH2PO3, sodium pyruvate, glucose, calcium galactonate, fetal calf serum, L-glutamine, an essential amino-acid, a nonessential amino acid, glutathione and water. The method and the used culture solution have excellent technical effects in the specification, for example, the method has a higher blastocyst hatching rate when the bovine in-vitro fertilized embryo is cultured.

The present invention discloses a vector, a cell and a method for improving bovine cloning efficiency on the basis of histone methylation modifying level. The vector includes a repressor expression vector and a repressor-operon-containing expression vector, and in the transfection, the repressor expression vector and the repressor-operon-containing expression vector are cotransfected into donor cells; and bovine KDM4B or bovine KDM4C gene is inserted into the repressor-operon-containing expression vector. Positive transfected cells as donor cells are injected into enucleated oocytes for electro-fusion, and successfully-fused bovine somatic cell cloned embryos are selected for culturing for 36h for induced expression of the bovine KDM4B and bovine KDM4C. Histone H3K9me3 epigenetic modifying level of the somatic cell cloned embryos in zygotic activation period can be effectively reduced, the development and quality of post-bovine somatic cell cloned blastocysts can be significantly improved, efficient and in-vitro production of the bovine somatic cell cloned embryos can be achieved, and birth rate of cloned cattle after embryo transplant can be significantly increased.

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.

A method of utilizing nuclear specific fluorescent staining and ovary being made to be completely transparent technique to observe blastocyst of rice includes using certain concentration of sodium hydroxide solution to carry out softening treatment on blastocyst of rice before staining then utilizing specific combination of nucleus fluorescent staining DAPI with nucleus to observe structure of cell and nucleus in blastocyst, enabling to use laser scan confocal microscope to observe internal structure of cell clearly.

A microbial prararation for increasing the absorptivity of plant to copper contains more useful microbes including scotch blastocyst, panicillium sp., etc. It is prepared by culturing them in liquid-state Ca's culture medium.

The invention discloses a culturing device, a culturing liquid and a culturing system for the external fertilization of human ovums and sperms and the early development of an embryo. The culturing device comprises at least two thermostatic culturing chambers, two small-sized air bottle chambers, a mixed gas filtering device, an air pressure protecting device, a mixed gas thermostatic preheating chamber, an air supplying pipe and the like, wherein each culturing chamber is a confined space with the volume of at least 100ml. The culturing liquid contains sodium chloride, magnesium sulfate, monopotassium phosphate, sodium bicarbonate, ethylenediamine tetraacetic acid, taurine, sodium pyruvate, L-lactic sodium lactate, glucose, gentamicin, alanine-glutamine dipeptide, 18 types of L-lactic amino acids except alanine and glutamine, and the like. When the culturing device and the culturing liquid are used, the external fertilization of the human ovums and sperms and the whole process of the development of the embryo before embryo implantation can be finished; the high-quality blastocyst forming rate and the embryo implantation rate are obviously increased.

The present invention provides a method for efficiently producing a germ-line chimeric animal and a method for conveniently producing a heterozygous animal and a homozygous animal. Specifically, a chimeric animal wherein germ cells are derived from introduced ES cells is produced by injecting embryonic stem cells (ES cells) into an early embryo (blastocyst-stage embryo) incapable of forming germ cells because of a genetic factor. A homozygote is obtained by crossing a female heterozygote with a male heterozygote obtained from the obtained chimeric animals. The thus obtained chimeric animal is extremely useful for conducting gene function analyses and the like.

The invention provides a method for improving quality of an external fertilization embryo of an oocyte of a sheep. The method relates to TrichostatinA (TSA) which is used for treating the oocyte of the sheep and can greatly improve the quality of the external fertilization embryo of the oocyte of the sheep, by adopting the method improving the quality of the external fertilization embryo of the oocyte of the sheep through TSA, after 168 hours, the blastocyst rate and the number of the obtained blastomeres are remarkably higher than those of the matched group, therefore, the method has better scientificity and practicability.

It is revealed that an organ such as pancreas can be regenerated by utilizing a fact that the deficiency of an organ is complemented by injecting an induced pluripotent stem cell (iPS cell) into a developed blastocyst in a blastocyst complementation method. Thus, the present invention has solved the above-described object. This provides a method for producing a target organ, using an iPS cell, in a living body of a non-human mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a different individual mammal that is an individual different from the non-human mammal.

Disclosed herein are devices, methods, systems and kits adapted for vitrification and / or reanimation of oocytes, embryos or blastocysts. The device includes a straw and a filter, wherein the straw comprises a lumen traversing through the straw and has a proximal section, a middle section and a distal section and wherein the filter is affixed in the straw and comprises a plurality of pores having a diameter smaller than the diameter of said oocytes, embryos or blastocysts but large enough to allow the passage of a fluid composition therethrough.