58 results about "Blastomere" patented technology

A method for determining viable normal blastomeres for implantation entailing labeling the blastomere with an antibody to hyperglycoslyated hCG and determining the binding of chromosomal probes directed to chromosomal regions of the chromosome.

The present invention provides a high efficiency method of deriving pluripotent mammalian stem cells from single, dissociated blastomeres harvested from preimplantation embryos.

The invention concerns a system and method for determining embryo quality comprising monitoring the embryo for a time period, said time period having a length sufficient to comprise at least one cell division period and at least a part of an inter-division period, and determining the length of the at least one cell division period; and/or ii) determining the extent and/or spatial distribution of cellular or organelle movement during the cell division period; and/or iii) determining duration of an inter-division period; and/or iv) determining the extent and/or spatial distribution of cellular or organelle movement during the inter-division period thereby obtaining an embryo quality measure. Thus, the selection of optimal embryos to be implanted after in vitro fertilization (IVF) is facilitated based on the timing, duration, spatial distribution, and extent of observed cell divisions and associated cellular and organelle movement.

Human non-embryonic adult totipotent and pluripotent stem cells are isolated in a simplified serum-free and feeder cell-free process. Most remarkably, certain stem cells, and especially BLSCs, are extremely small, fail to exclude trypan blue, but are nevertheless able to proliferate from even high dilutions. Therefore, so obtained stem cells can be used to prepare true monoclonal stem cell populations, which are useful in numerous uses, including therapeutic, prophylactic, diagnostic, and research uses.

The invention relates to the field of biotechnology, in particular to a method for constructing a lethal gene systemic knockout mouse model with a CRISPR/Cas9 system. The method includes the followingsteps that 1, an sgRNA sequence for efficiently identifying a lethal gene PAM region after knockout is designed; 2, mRNA or protein of Cas9 is mixed with the sgRNA designed in step 1, microinjectionis performed on any blastomere cell in a mouse two-cell embryo with the mixture, embryo transplantation is performed after injection, and strain gene identification is performed to obtain a chimeric positive founder with the lethal gene knocked out; 3, the positive founder and a wild-type mouse are mated to generate an F1 generation, a heterozygous F1-generation mouse with the lethal gene systemically knocked out is finally obtained after gene identification, and construction of the mouse model capable of realizing passage propagation is completed. Compared with the prior art in which a lethalgene systemic knockout mouse model is constructed by ES cell gene targeting, the method in the technical scheme has the advantages that the operation steps are simple, the operation difficulty is low, the production cycle is short, and the cycle is only about 4 months.

The invention relates to a method for producing transgenic buffalo embryos by applying intracytoplasmic sperm injection (ICSI) mediation. The method comprises the following steps of: 1, incubating buffalo sperms and transgenic deoxyribonucleic acid (DNA) together; 2, performing intracytoplasmic microinjection on the buffalo sperms; and 3, cultivating and identifying the transgenic buffalo embryos. By the method of the invention, the transgenic buffalo embryos are obtained successfully. In 102 injection spawns of the transgenic buffalo embryos, 61.8 percent of the injection spawns are subjected to cleavage, 17 blastospheres are obtained in embryos expressed by blastomere serving as enhanced green fluorescent protein (EGFP), 11 blastospheres express the EGFP, and the transgenic positive rate of the blastospheres is up to 64.7 percent; and after one of the transgenic embryos is cultured in vitro for a long time, the expression of the EGFP can be still seen during 2 to 7 days, so the method makes the important progress for producing transgenic buffalos further.

The invention discloses a method for improving pig nucleus transplantation efficiency, which belongs to the field of breeding or inseminating methods. The method comprises the following steps of: processing a donor cell by a small molecule medicament Calyculin A and inducing donor chromatin condensation, methylation rise and acetylation reduction, wherein a somatic nucleus expresses genetic modified zero resetting; then carrying out nucleus transplantation and constructing reconstructed embryos; and selecting optimal concentration of 10nM and the processing time of 30 minutes by an optimizing condition. The invention improves the ectogenesis blastocyst rate of a pig clone embryo by 9.7 percent, increases 23.78 blastomeres and lays the foundation for improving the production efficiency of clone animals.

The present invention provides a method for parthenogenetic activation by injection and subsequent removal of a sperm into a mammalian cell, for example, an oocyte, an embryo, a blastomere, an inner cell mass cell, or a morulae cell.

The invention discloses an embryo quality comprehensive evaluation device based on deep learning. The device comprises a computer memory, a computer processor and a computer program which is stored inthe computer memory and can be executed on the computer processor, an embryo quality comprehensive evaluation model is stored in the computer memory, and when the computer processor executes the computer program, the following steps are realized: receiving an embryo image; identifying and segmenting a regional image of the cleavage ball through a cleavage ball target detection module; using the cell counting module to count cells in the regional image, and obtaining the development stage of the cleavage ball; using the embryo quality analysis module to score the development condition of the cleavage ball in the regional image; and in the prediction module, scoring the implantation possibility of the cleavage ball according to the regional image, and obtaining a prediction result of the implantation success rate of the cleavage ball by integrating the scores of the development conditions. The prediction result can assist a doctor in predicting the success rate of embryo implantation.

The invention discloses a cloning method of the animal somatic cells. The cloning method of the animal somatic cells is that tetraploid blastomere in 8-16 cell stage which is obtained from vitro fertilization is transferred into a somatic cloned embryo in 8-16 cell stage to get a compound embryo; the compound embryo is cultured to get a cloned animal. The invention initiatively changes the sourceof trophoblast cell; the tetraploid blastomere obtained from vitro fertilization is put into the somatic cloned embryo in 8-16 cell stage, which can be developed into trophoblast cell; then the placenta quality is improved, the development condition for the fetus is promoted, the pregnancy rate of the cloned embryo is increased, the abortion rate and the dystocia rate is decreased and the cloningefficiency of the somatic cell is greatly improved. As the fetus is constituted by ploid cells, the tetraploid blastomere can only develop into the placenta without the possibility of developing intofetus. Therefore, the method does not affect the individual cloned animal.

We developed a generic approach to type genome-wide single nucleotide polymorphisms in single human cells and to reconstruct for the first time genome-wide haplotypes of single- or dual-cell derived genotypes. Proof-of-principle is delivered for EBV-transformed lymphoblastoid cells as well as human blastomeres. To this end, multiple displacement amplified DNA samples of single cells were hybridized to Affymetrix 250K SNP-arrays. Different algorithmic designs were subsequently developed to assess from the single-cell derived SNP-probe intensities the sequence of syntenic alleles and to pinpoint accurately the majority of parental homologous recombination sites across the entire genome using a linkage-based approach. This included the development of algorithms that rectify a large part of the discrepant allelic assignments in raw single or dual-cell derived haplotypes. This method to infer genome-wide haplotypes from the analysis of only one or two cells has tremendous applicative value. It has the capacity to revolutionize not only genetic diagnosis of preimplantation in vitro fertilized human embryos in the clinic, but also animal breeding programs by enabling genome-wide quantitative trait loci selection at the embryonic level. In addition, it allows to further scrutinize drivers of haplotype diversity, mainly meiotic homologous recombination as well as somatic (homologous) recombination processes that occur often during (human) tumorigenesis.

The invention belongs to the field of biotechnologies, and relates to a method and culture medium for culturing transgenic animal embryonic cells or transgenic animals. The method comprises the steps that animal fertilized eggs obtained after gene modification are cultured to the four-cell stage, a zona pellucida is removed, then plasmodesmata are removed, four-cell embryos are separated into four single blastomeres, then the single blastomeres are put back into the zona pellucida with cytoplasm removed, in-vitro culture is carried out, and homozygous gene modified embryos are obtained efficiently. The optimized culture medium is provided at the same time. Based on the embryo blastomeres separation and independent culture technology, blastospheres can be successfully obtained in an optimized embryo culture system, and the efficiency is 30.33%. In addition, the homozygosis targeting efficiency of the obtained regenerated embryos is relatively improved obviously relative to that of unsplit developing embryos, and the homozygous mutation efficiency reaches up to 55.80%.

The invention discloses a cleavage ball cleavage state intelligent detection method and system. The method comprises the following steps: collecting an embryo image in periods D1 to D3 and recording ashooting time point; inputting the embryo image and the time point thereof into a cleavage ball number prediction network model for prediction; calculating a cleavage globule division index F, and judging whether the embryo is normally divided in the cleavage stage or not; and outputting the predicted number of the cleavage balls in each embryo image, the cleavage ball cleavage curve and the cleavage state judgment result of the embryo in the cleavage stage as a detection report. According to the detection method provided by the invention, the whole process of the cleavage ball development ismonitored; the number of the cleavage balls and cleavage ball cleavage points can be accurately detected; a doctor can rapidly and visually observe and analyze the whole cleavage process of the cleavage ball, time and money cost are saved for the doctor and a patient, the prediction accuracy of the number of embryos of the cleavage ball number prediction network model reaches 93%, and the accuracy is far higher than that of the traditional image processing method.

The invention relates to the technical field of biology and in particular relates to a method for constructing a knock-out mouse model to analyze a lethal gene function by utilizing a CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9) system. The method comprises the following steps: 1) designing an sgRNA (small guide Ribonucleic Acid) sequence forefficiently identifying a PAM region of a knocked-out lethal gene; 2) mixing Cas9 or protein of mRNA (micro Ribonucleic Acid) and sgRNA designed in step 1); carrying out microinjection on any one blastomere cell in a mouse two-cell embryo through a mixture; after injection is finished, transplanting the embryo and identifying strain genes to obtain a chimera positive mouse with the knocked-out lethal gene; 3) analyzing the function of the lethal gene in a tissue organ by utilizing a positive founder mouse. According to the technical scheme provided by the invention, compared with the prior artof specifically knocking out tissues based on a Cre/LoxP system, the method has a short manufacturing period and the period is only about 3 months.

The invention discloses a grade classification method for evaluating an in-vitro fertilization treatment embryo based on cleavage behaviors. According to the invention, on the premise that a human embryo cell is neither interfered nor damaged, data of dynamic observation on a human embryo is obtained by employing a time-lapse imaging technology and abnormal cleavage behaviors of the human cleavage-state embryo of in-vitro fertilization are collected and described in detail, and divided into 7 types. Based on the order of severity of abnormal cleavage, blastomere range affected by the abnormal cleavage behaviors, and the generation period of the abnormal cleavage behaviors, the qualitative embryo grade classification method with little subjectivity is established, for effectively screening out the cleavage-state embryo with maximum blastocyst development potential.

The invention relates to a full-automatic single cell tube filling operation system for preimplantation genetic diagnosis. The full-automatic single cell tube filling operation system comprises a transparent bottom plate, a transparent rotating disc, a transmission wheel, a motor, a rotating vertical rod, a rotating transverse rod and an operation arm, wherein the operation arm is used for grasping blastomere; the transmission wheel is mounted on the transparent bottom plate through a rotating shaft; the rotating shaft is connected with the motor; the transparent rotating disc is mounted on the transparent bottom plate through a fixing shaft and a bearing; the transparent rotating disc is meshed with the transmission wheel through teeth; the rotating vertical rod is movably mounted on the transparent bottom plate; one end of the rotating transverse rod is movably mounted at the upper end of the rotating vertical rod; a sliding groove is formed in the other end of the rotating transverse rod; the operation arm is mounted inside the sliding groove through a sliding block; the operation arm is positioned above the transparent rotating disc; telescopic support feet are further arranged at the bottom end of the transparent bottom plate. The full-automatic single cell tube filling operation system does not need to be fixed on a certain device, can be flexibly moved and is used together with various stereoscopes; the tube filling accuracy can be improved, the success rate of single cell tube filling operation can be increased, a great deal of labor and time is saved, and the working efficiency is improved.

The invention discloses an editing method aiming to female genes. The editing method comprises the following steps of acquisition of parthenogenetic fertilized eggs, editing of the parthenogenetic fertilized eggs, separation of edited parthenogenetic embryo blastomeres, reconstruction of parthenogenetic embryos, and identification of an editing result. The method provided by the invention can indirectly realize a function of performing precise editing on mammal ovum, can ensure the final gene editing effect and is beneficial to conversion of a gene editing technology to clinical medicines.

A primary mammalian cell which may be infected with a pathogen is cultivated under reduced oxygen to improve susceptibility to infection with a pathogen, pathogen propagation, and/or to change or maintain a degree of differentiation of the cell in a predetermined manner. In preferred aspects, the cell is a hepatocyte, optionally infected with HCV, or a blastomere from a single-cell biopsied blastocyst. Such biopsied cell may then be expanded to a population of stem cells under concurrent cultivation of the blastocyst, which is then used for IVF implantation.