414 results about "Embryo culture" patented technology

We describe a method for interrogating the content and primary structure of DNA by microarray analyses and to provide comprehensive genetic screening and diagnostics prior to embryo transfer within an IVF setting. We will accomplish this by the following claims: 1) an optimized embryo grading system, 2) a less invasive embryo biopsy with reduced cellular contamination, 3) an optimized DNA amplification protocol for single cells, 4) identify aneuploidy and structural chromosome abnormalities using microarrays, 5) identifying sub-telomeric chromosome rearrangements, 6) a modified DNA fingerprinting protocol, 7) determine imprinting and epigenetic changes in developing embryos, 8) performing genome-wide scans to clarify / diagnose multi-factorial genetic disease and to determine genotype / haplotype patterns that may predict future disease, 9) determining single gene disorders with or without a known DNA mutation, 10) determining mtDNA mutations and / or the combination of mtDNA and genomic (nuclear) DNA aberrations that cause genetic disease.

The invention provides a method for knocking out p66shc in a pig embryo by utilizing a CRISPR-Cas9 gene editing method. The method comprises the following concrete operation steps: (1) designing an Oligo DNA sequence; (2) connecting Oligo; (3) transforming and amplifying a constructed plasmid; (4) verifying knock-out efficiency by plasmid transfected cells; (5) constructing a CRISPR-Cas9 system of the p66shc gene; (6) producing the pig embryo with the p66shc gene knocked out; and (7) verifying a p66shc gene knock-out result. By adopting the scheme of the invention, the ROS level in an in vitro embryo culture process is effectively reduced, so that in vitro production efficiency of the pig embryo is improved by reducing damage and the like in the in vitro embryo culture process, and finally, an effective in vitro production system of the pig embryo is constructed.

A culture dish (1) comprising a basic structure (2) with a number of macro wells (4) for culturing oocytes and embryos, the culture dish further comprising a lid (3) with a movable part (8), the movable part (8) having an opening (11) adapted to the size of the macro wells (4), the movable part (8) being movable between a first position where all macro wells are closed by the lid (3) and further positions where for each further position (11) the opening (11) is aligned with one of the macro wells (4) to allow access to this macro well (4) through the opening (11) in the lid (3).

The invention provides a culture fluid exclusively used in cow in-vitro fertilization embryos. The culture fluid contains 109.0mM-110mM of NaCl, 2.9mM-3.1mM of KCl, 26.0mM-26.5mM of NaHCO3, 0.5mM-1.0mM of MgCl2.6H2O, 1.0mM-1.3mM of KH2PO3, 0.4mM of sodium pyruvate, 1.5 mM of glucose, 5mM of galacturonic calcium, 10v / v% fetal bovine serum, 1mM of L- glutamine, 2v / v% essential amino-acid, 1v / v% nonessential amino acid and 1mM-10mM of glutathione. The cow in-vitro fertilization embryos are placed into the culture fluid to carry out in-vitro cultivation, and results are shown to be obviously superior to those of a control group which is not added with GSH (glutathione), and therefore, developmental rate of blastula and embryo quality are improved, cost for producing embryos in vitro is lowered, experimental basis is provided for applying a cow IVF (in vitro fertilization) technology to practice, and a genetic breeding process can be greatly accelerated.

The invention provides a culture fluid exclusively used in cow in-vitro fertilization embryos. The culture fluid contains 109.0mM-110mM of NaCl, 2.9mM-3.1mM of KCl, 26.0mM-26.5mM of NaHCO3, 0.5mM-1.0mM of MgCl2.6H2O, 1.0mM-1.3mM of KH2PO3, 0.4mM of sodium pyruvate, 1.5 mM of glucose, 5mM of galacturonic calcium, 10v / v% fetal bovine serum, 1mM of L- glutamine, 2v / v% essential amino-acid, 1v / v% nonessential amino acid and 1mM-10mM of glutathione. The cow in-vitro fertilization embryos are placed into the culture fluid to carry out in-vitro cultivation, and results are shown to be obviously superior to those of a control group which is not added with GSH (glutathione), and therefore, developmental rate of blastula and embryo quality are improved, cost for producing embryos in vitro is lowered, experimental basis is provided for applying a cow IVF (in vitro fertilization) technology to practice, and a genetic breeding process can be greatly accelerated.

The invention discloses a three-step method for producing a 'dragon head phoenix tail' high-quality officinal dendrobium stem material. The method comprises three steps of embryo culture for direct seedling formation, plug plantation of seedlings in a greenhouse, and culture of a semi-mature seedling epiphytic trunk; and an officinal dendrobium stem production technology is simplified, seedling breeding time is shortened, and the quality and transplanting survival rate of test tube seedlings of officinal dendrobium stem are improved. In the method, the production time of the test tube seedlings is 6 to 7 months, but the conventional test tube seedling breeding time is 10 to 12 months. An officinal dendrobium stem medicinal material prepared by the method has a thick and short stem, the content of polysaccharide from Dendrobium officinale is 36 to 48 percent, and the officinal dendrobium stem medicinal material has a 'dragon head phoenix tail' characteristic in external shape and is a necessary raw material for processing 'dragon head phoenix tail' Tiepi Fengdou. A semi-mature seedling epiphytic trunk technology in the method is suitable for farmers in mountainous areas to cultivate the officinal dendrobium stem, and is low in investment and high in yield; the farmers do not need to build greenhouses and only need to purchase plug seedlings; and the cultivation technology is easy to grasp and is a practical technology for servicing 'Three Rural Issues'.

The invention relates to a method for improving in-vitro production efficiency of buffalo embryos, comprising the following steps of: respectively adding 10 muL / mL of insulin-transferrin-selenium (ITS) and 20 muL / mL of insulin-transferrin-selenium (ITS) to an oocyte maturing solution and a culture solution, and then carrying out in-vitro maturation and in-vitro culture. The method indicates that the in-vitro maturation rate of buffalo oocyte is effectively improved by adding 10 muL / mL ITS to the maturing solution, and the exhaustion ratio of a first polar body is up to 65.61%; and adding 20 muL / mL of ITS to the embryo culture solution is beneficial to the growth of early embryos of a fertilized buffalo, and the growing rate of a blastaea is up to 29.93%. the method has the advantages of improving the in-vitro maturation rate of the buffalo oocyte and the growing rate of the blastaea, thereby improving the in-vitro production efficiency of the buffalo embryos and providing a large number of high-quality embryos for embryo transplantation.

A method for determining embryo quality involving measuring soluble HLA-G levels present in the embryo culture medium at least 44-46 hours post-fertilization is provided. Culture media and in vitro fertilization programs employing same are also provided.

The invention relates to a bovine oocyte in-vitro fertilization and embryo culture method, and a transport culture solution. On one hand, the transport culture solution comprises glycine, alanine, arginine hydrochloride, aspartic acid, cystine dihydrochloride, glutamic acid, L-glutamine, histidine, threonine, tryptophan, tyrosine, valine, ascorbic acid, biotin, choline chloride, calcium pantothenate, folic acid, menadione and the like. The bovine oocyte in-vitro fertilization and embryo culture method comprises the following steps: oocyte collection and in-vitro maturation, in-vitro fertilization, and embryo in-vitro culture and preservation. The method and the related transport culture solution have excellent technical effects as described in the specification.

A container or dish used for the micro manipulation, micro injection, biopsy and fertilization of oocyte and embryo culture. The dish allows the user to more readily perform procedures used to fertilize oocyte, such as intracytoplasmic sperm injection (ICSI), biopsy embryos and perform additional procedures in used in assisted reproductive techniques (ART), human reproduction and in vitro fertilization (IVF) techniques. The invention will allow ease in use, the reduction in the number of micro tools used in the procedure, as opposed to conventonal dishes and procedures, and will allow the user to more readily locate the oocytes and embryos to be handled and worked on. The inventon will add repetitiveness, consistency and may result in better results and outcome of the procedures. The invention will also give the user a more ergonomically correct dish for these types of procedures and related protocols.

The invention provides a cultivation method of cross-bred wagy. The method comprises the steps of culturing cross-bred wagy embryo in vitro and transplanting the cultured embryo into uterus of receptor cow to breed, wherein the embryo in-vitro culture method comprises the following steps: (1) taking an ox with 99.9% of Japanese wagy hereditary characters as a paternal line, and Holstein cow as a maternal line, respectively extracting semen of the paternal line and ovocyte of the maternal line; (2) culturing the ovocyte in ovocyte follicle fluid; (3) performing in-vitro fertilization on mature ovocyte in an in-vitro fertilization culture solution; (4) culturing the fertilized ovum in the embryo culture solution; and (5) vitrifying the in-vitro embryo. The in-vitro cultured embryo is transplanted into the uterus of receptor cow by a bilateral uterine lining embryo transplantation method. At last, the cross-bred first filial generation wagy new strain with characters of Japanese wagy and Holstein cow is cultivated, and has high table purpose value and good production performance.

The invention relates to a seed germination technology, and provides a method for Louisiana iris hybrid seed germination. The method comprises the following steps: immersing the matured Louisiana iris hybrid seeds in sterile water for 24 hours, sucking water for satiation and disinfecting, stripping a complete embryo for inoculating in a solid medium for in-vitro culture in dark, inoculating to the solid medium of a same component after the embryo is germinated, and then illuminating for culturing, and acquiring the hybrid seed. The method of the invention employs an embryo culture technology to perform in-vitro culture on the hybrid seeds in period of dormancy, the hybrid seeds can break the dormancy, and the embryo enables rapid expansion and elongation in the medium. Compared with a routine sowing method, the germination rate in the invention increases by 60%, the germination time shortens by 12 days, seedling emergence time shortens by 20 days, the operation is simple, and the workload is not increased.

The invention belongs to the field of haploid breeding technology and discloses a wheat double haploid production method by wheat-and-maize distant hybridization. The main technological characteristics of the method comprise steps of planting, induction of haploid embryo, dissection of haploid embryo, respective isolated culture of small embryo and big embryo into haploid plantlet, vernalization,chromosome doubling, transplantation and harvesting. The big embryo is cultured in big embryo culture medium. The small embryo is firstly cultured in small embryo culture medium and then cultured in basal medium. Small embryo culture medium has a higher sugar concentration, a higher osmotic pressure, richer kinds of amino acids, higher vitamin contents and contains trace of growth regulation factors with the germinating and seedling rate of small embryo being over 75%. By low-temperature vernalization, isolated cultured tissue culture seedlings are directly carried out chromosome doubling andplanting with the doubling treatment efficiency being over 89%. The method provided by the invention is adopted to improve seedling rate of haploid embryo and survival rate and success rate of chromosome doubling, simplify wheat breeding program, increase breeding efficiency and shorten breeding cycle, and has apparent effects on accelerating cultivation of fine varieties.

The invention relates to an application of zebra fish to testing water quality and toxicity and a method for applying zebra fish to test water quality and toxicity. The method is characterized by comprising the following steps of: preparing an embryo culture solution, then adding a water source to be tested to the embryo culture solution, using the embryo culture solution to which the water source to be tested is added to culture zebra fish embryos and then observing and recording the developmental aberration rates and mortality of the zebra fish embryos in 12-24 hours to judge the toxicity of the water source to be tested toward the embryos, wherein the ratio of the volume of the added water source to be tested to the volume of the embryo culture solution is 1:3; the embryo culture solution is directly prepared from alkalescent water with pH value being 6.7-8.5; and the embryo culture solution contains 5mM of NaCl, 0.17mM of KCl, 0.43mM of CaCl2 and 0.33mM of MgSO4. The method has the following beneficial effect that the embryo culture solution prepared from the alkalescent water ensures that the zebra fish embryos can quickly test the comprehensive toxicity of the water source to be tested, thus shortening the detection period.

The invention relates to a method for applying zebra fish to test toxicity of an organic solvent. The method comprises the following main steps of: preparing an embryo culture solution, then adding the organic solvent to the embryo culture solution, ensuring that the concentration of the organic solvent in the embryo culture solution reaches the concentration needed by measuring, using the embryo culture solution to which the organic solvent is added to culture zebra fish embryos and then observing and recording the developmental aberration rates and mortality of the zebra fish embryos in 6-12 hours to judge the toxicity of the organic solvent toward the embryos, wherein the embryo culture solution is directly prepared from alkalescent water with pH value being 6.7-8.5; and the embryo culture solution contains 5mM of NaCl, 0.17mM of KCl, 0.43mM of CaCl2 and 0.33mM of MgSO4. The method has the following beneficial effect that the embryo culture solution prepared from the alkalescent water ensures that the zebra fish embryos can quickly test the toxicity of the organic solvent, thus shortening the detection period.

The invention relates to a modified zebra fish drug metabolism modeling method, which specifically comprises the steps of (1) enriching metabolites by virtue of a zebra fish metabolism model; (2) separating and analyzing the enriched metabolic components of the zebra fish in the last step by using modern chromatography or the hyphenated technique thereof, capturing or knocking out target components or metabolites; (3) collecting zebra fish germ cells; (4) putting zebra fish embryos in three days after fertilization in a culture plate which holds an embryo culture medium and administrating a drug; (5) dyeing the bones of the zebra fish; and (6) detecting and analyzing the dyed bones of the zebra fish. According to the invention, an adolescent zebra fish osteoporosis model is applicable to in-vivo effective evaluation of anti-osteoporosis activity of Chinese herbal medicinal ingredients such as trace ingredients; an adult zebra fish metabolism model is simple in metabolites and efficient in enrichment; the modern chromatography or the hyphenated technique thereof provide powerful guarantee for the separation and analysis of the complex Chinese herbal medicinal system and the metabolic components. The model obtained by the method has the outstanding advantage that the separation and analysis of Chinese herbal medicinal original-form ingredients and the metabolic components are integrated with the in-vivo anti-osteoporosis activity evaluation, and simple and efficient.

The invention discloses a method for increasing breeding efficiency of wild rice and cultivated rice distant hybridization embryo rescue, belongs to the field of breeding by the biological technique, and aims at reducing the alcohol disinfecting time and reducing the alcohol concentration as well as creating foundation for the survival rate of hybridization seedlings. The method is that the MS of an embryo culture is improved; the concentration of KNO3, NH4NO3, KH2PO4 and MnSO4.4H2O is reduced; meanwhile, CuSO4.5H2O, CoCl2.6H2O, niacin, pyridoxine hydrochloride and thiamine hydrochloride are not added; the hardening-seedling technology is innovated; when in hardening seedling of the plants, the hardening-seedling treatment with convenient operation and good effect is carried out, so that the survival rate of the seedlings can be greatly increased. According to the operation of the method, the operation of embryo rescue is simplified; the whole culture costs 30 days only, so that the culture cycle can be greatly reduced; meanwhile, the seedling rate can be greatly increased and up to 94%; therefore, the breeding efficiency of the wild rice and cultivated rice distant hybridization embryo rescue can be increased.

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 invention provides a method of performing scRRBS (single-cell reduced-representation bisulfite sequencing) on an embryo culture solution. According to the method provided by the invention, medicalwaste (blastula culture solution) generated by 'test-tube baby' operation is adopted as a raw material, double analysis can be performed on the chromosome aneuploidy status and DNA methylation statusof the embryo, development potential of the embryo is evaluated from a brand new angle of epigenetics and reaction between the embryo and the culture environment, new reference is provided for selecting a 'correct' embryo in assisted reproduction, and powerful support is provided for increasing the success rate of the period of the test-tube baby.

The invention relates to a mature wheat embryo culture and regeneration method. Regeneration culture mediums are based on MS basic culture mediums. In the regeneration culture mediums, the content of maltose is 40g / L, the content of MES is 1.95g / L, the content of KT is 5mg / L, the content of NAA is 0.1mg / L and the pH is 5.8. The specific method comprises the steps of selecting mature and full wheat seeds, soaking the seeds in 70% alcohol solution for 5min, sterilizing the seeds in 0.1% HgCl2 solution for 20min, washing the seeds for at least four times by using sterile water and soaking the seeds in the sterile water for 19-24h; removing the buds and the roots of the mature embryos of the seeds, longitudinally cutting the mature embryos, putting the cut mature embryos into induction culture mediums to culture in the dark at 23-25 DEG C, conducting subculture once for 10-14 days and conducting induction culture for totally 20-28 days to obtain mature wheat embryo calluses; and then transferring the mature wheat embryo calluses into the prepared regeneration culture mediums to culture in the light at 23-25 DEG C for 25-30 days and inducing the mature wheat embryo calluses to be differentiated to form regenerated seedlings.

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 relates to the field of cross breeding of plants and particularly relates to an interspecific hybrid embryo rescuing and seedling forming method of sweet cherries in southern China and Chinese cherries. The method combines advantages of European sweet cherries and the Chinese cherries, and takes the 'European sweet cherries' as a female parent and the 'Chinese cherries' as a male parent to carry out interspecific distant hybridization. Aiming at the abortion problem of a distant hybrid embryo, the hybrid embryo is subjected to embryo rescuing. A proper sampling period of the embryo rescuing of each hybrid combination, concentrations and ratios of respective exogenous growth regulators for hybrid young embryo germination, subculture propagation and rooting culture, and a proper acclimatization and transplanting method are determined. By applying an interspecific hybrid embryo rescuing system of the sweet cherries in southern China and the Chinese cherries, which is established by the technology, the embryo germination rate reaches 80.00 percent, the multiplication coefficient of subculture hybridized new shoots reaches 5.0, the rooting rate of rooting culture is 90.91percent, the growth vigor of embryo culture seedlings is good and the seedlings grow robustly and have green leaves, and the acclimatization and transplanting survival rate reaches 77.27 percent.

The invention provides a method for cultivating hybrid plants of far-source lilies, which overcomes the problem that far-source hybrids are not easy to be affinitive in the prior art. The method performs in vitro culture to an ovary and an anther, adopts an unconventional pollination mode of cutting off the ovary for in vitro pollination, and then performs embryo culture so as to obtain a hybrid plant. Hybrid seedlings are obtained by use of the method, which proves that the method successfully overcomes the problem that the far-source hybrids are not affinitive, can well improve seed setting rate, and is worthy to be popularized and applied.

The invention discloses a method for producing virus-free atractylodes macrocephala koidz seedlings. The method comprises the following steps: 1, pre-treating atractylodes macrocephala koidz seeds and aseptically sprouting the embryos atractylodes macrocephala koidz seeds, to be specific, inoculating atractylodes macrocephala koidz embryos into a primary culture medium comprising 1 / 2MS, 20g / L sucrose and 6g / L agar powder; 2, performing proliferation culture, wherein a proliferation culture medium comprises MS, 1.0mg / L 6-BA, 0.2mg / L NAA, 30g / L sucrose and 6g / L agar powder; 3, detecting viruses, to be specific, rejecting virus-containing atractylodes macrocephala koidz seedlings, and continuing to perform proliferation culture on the virus-free atractylodes macrocephala koidz seedlings; 4, performing rooting culture, to be specific, uprightly inserting rootless atractylodes macrocephala koidz tissue culture seedlings which are cultured to about 4cm into a rooting culture medium comprising 1 / 2MS, 1.0mg / L NAA, 20g / L sucrose and 6g / L agar powder; 5, performing acclimatization and transplantation. According to the method, the formulae of the primary culture medium and the rooting culture medium for atractylodes macrocephala koidz are obtained by single-factor tests, the formula of the proliferation culture medium for the atractylodes macrocephala koidz tissue culture seedlings is obtained by an orthogonal design, and most importantly, the virus-free atractylodes macrocephala koidz seedlings are obtained in an embryo culturing way by utilizing the fact that embryo parts are virus-free.

The disclosure relates to the use of culture media containing thyroid hormones or analogs thereof, and includes methods and uses thereof for embryo culture, embryo production, embryo maturation, improved survival of embryos and improved viability of embryos post cryopreservation.

An integrated automated system comprising a microfluidic cassette, and methods of use thereof, for intracytoplasmic sperm injection assisted fertilization. The microfluidic cassette and the integrated automated system provides a complete set-up of human gametes for assisted in vitro fertilization, including proper cell stage recognition, gamete propulsion via microfluidic currents, microinjection of a spermatozoon into an oocyte, and subsequent embryo culture and monitoring, thus allowing widespread distribution of in vitro insemination by favoring affordability.

The invention relates to an application of zebra fish to testing safety of a safe substance and a method for applying zebra fish to test safety of the safe substance. The method is characterized by comprising the following steps of: preparing an embryo culture solution, then adding the safe substance to be tested to the embryo culture solution, using the embryo culture solution to which the safe substance to be tested is added to culture zebra fish embryos and then observing and recording the developmental aberration rates and mortality of the zebra fish embryos in 4-72 hours to judge the toxicity of the safe substance to be tested toward development of the zebra fish embryos, wherein the addition of the safe substance to be tested ensures that the concentration of the substance to be tested in the culture solution is distributed in echelon; the embryo culture solution is directly prepared from alkalescent water with pH value being 6.7-8.5; and the embryo culture solution contains 5mM of NaCl, 0.17mM of KCl, 0.43mM of CaCl2 and 0.33mM of MgSO4. The method has the following beneficial effect that the embryo culture solution prepared from the alkalescent water ensures that the zebra fish embryos can quickly test the comprehensive toxicity of the safe substance to be tested, thus shortening the detection period.

The invention provides a calf in vitro embryo culture solution containing cysteamine. The early-stage embryo in vitro development culture solution is used to culture the calf in vitro embryo and can obtain higher blastaea developmental rate.

The invention relates to a bamboo blank culture medium, which uses MS as basic culture medium, with 3.1 or 4mg / L growth adjuster, nutriment at 3%, solidifier at 0.25%, and the pH value of medium is 5.7. The inventive production comprises that selecting seeds, absorbing water and activating, disinfecting, selecting blank, cultivating in tube, transplanting, and planting in field. The invention can improve the bamboo kind.

The invention provides an embryo culture medium for Lei bamboo (Phyllostachys praecox), which comprises the following components by weight part: MS salts, MT vitamins, 0.3 mg / L or 1 mg / L or 3 mg / L of BA (i.e. benzylamino purine), 0.1 mg / L of NAA (i.e. alpha-naphthaleneacetic acid), 0.1 mg / L of 3,5,6-trichloro-4-aminopicolinic acid, 0.001 mg / L of TDZ (i.e. thidiazuron), 20 g / L of glucose, 100 mg / L of inositol, 10 mg / L of adenine sulfate, and 0.5 g / L of malt extract. The pH value of the culture medium ranges from 5.0 to 6.0. The method of tissue culture and seedling cultivation using the culture medium comprises the following steps: collection and selection of explants, cleaning and sterilization of seeds, and extraction, inoculation, culture, sub-culture or transplanting of the embryos. The culture medium and the seedling tissue culture method are excellent in the cultivation of the Lei bamboo as the medium-diameter running bamboo with high economic value, lay the foundation for researching and bioengineering of Lei bamboo, and realize the rapid and sufficient supply of Lei bamboo seedlings.