58 results about "Spermatocyte" patented technology

Improved flow cytometer system particularly adapted to use for sex-selected sperm sorting include enhanced sheath fluid and other strategies which minimize stress on the sperm cells, including a 2.9 percent sodium citrate sheath solution for bovine species and a hepes bovine gamete media for equine species. Improved collection systems and techniques for the process are described so that commercial applications of sperms samples as well as the resulting animals may be achieved.

The present invention provides a method of cryopreserving sperm that have been selected for a specific characteristic. In a preferred embodiment, the method is employed to freeze sex-selected sperm. Although the cryopreservation method of the invention can be used to freeze sperm selected by any number of selection methods, selection using flow cytometry is preferred. The present invention also provides a frozen sperm sample that has been selected for a particular characteristic, such as sex-type. In preferred embodiments, the frozen sperm sample includes mammalian sperm, such as, for example, human, bovine, equine, porcine, ovine, elk, or bison sperm. The frozen selected sperm sample can be used in a variety of applications. In particular, the sample can be thawed and used for fertilization. Accordingly, the invention also includes a method of using the frozen selected sperm sample for artificial insemination or in vitro fertilization.

The present invention provides methods for preparing cells with highly condensed chromosomes, such as sperm, and methods for detecting and quantifying specific cellular target molecules in intact cells. Specifically, methods are provided for detecting chromosomes and chromosomal abnormalities, including aneuploidy, in intact cells using fluorescence in situ hybridization of cells in suspension, such as sperm cells.

Methods and apparatus are disclosed for processing sperm cells to accomplish preservation for future use while minimizing the adverse effects of such preservation. Sperm cells may be collected from a male animal and subjected to a first preservation step, including potentially a first cryopreservation step. Preserved sperm may then be revived, including potentially by thawing, and treated by any of various processing steps to mitigate the adverse effects of preservation. Treated sperm may then be subjected to a second preservation step, including potentially a second cryopreservation step, perhaps enabling a delayed use of the sperm at a future time.

Disclosed are sperm cells or embryos comprising a labeled oligonucleotide bound to a gender-specific repeat sequence. Methods for separating sperm cells or embryos containing a labeled oligonucleotide from sperm cells not containing the labeled oligonucleotide produce gender-enriched sperm cell fractions. The separated fractions are useful in producing offspring of a predetermined sex.

In various aspects, the present invention provides novel and effective methods and kits for the isolation of sperm and sperm DNA from samples having at least one other cell type, or having the DNA of one other cell type. More specifically, a process for isolating sperm DNA from a mixture of sperm and non-sperm cells by filtration is provided. DNA from non-sperm cells is solubilized by selective lysis, and the intact sperm are retained on a filter, washed, and then treated in situ with a reducing agent to solubilize the sperm DNA. Significantly, centrifugation steps are not required, the process can be easily automated, and can be performed on many samples in parallel. The novel methods and kits are based on filtering selectively solubilized samples through filters that retain sperm by virtue of having uniform pore diameters that: are smaller than sperm, large enough to allow passage of cell debris and solubilized DNA; and are stable to pressure. The inventive methods and kits have broad utility, particularly in the forensic art.

The invention discloses novel growth hormone releasing hormone analog peptides and an application thereof in preparing medicines for treating infertility. Experiments discover that 2D, 2E or 2F peptide has obviously high hypophysis GH releasing activity and hypophysis hormone releasing specificity. Tested by the conjugation reaction of an in-vitro GHRH dimer peptide and a hypophysis GHRH receptor, 2D, 2E and 2F dimer peptides have extremely high hypophysis receptor binding activity, wherein the 2F dimer peptide has the maximum binding activity. With the 2F peptide as the representative, infertility model treatment finds out that, compared with a normal saline group and a pure cyclophosphamide control group, in the 2F dimer peptide group, the spermatocytes and the spermatogonia in the seminiferous tubules are obviously increased, the seminiferous tubule cells are arranged in order and large in volume, the cavities of the seminiferous tubules are reduced and even disappear, and dose dependency is shown. All the facts indicate that the GHRH peptides with the 2F peptide as the representative have an obvious effect of stimulating the proliferation and the maturation of spermatogonia / oogonia, thereby promoting reproduction; and as a result, the novel growth hormone releasing hormone analog peptides can be applied to medicines for treating infertility.

The present invention is directed to a vector and its use to generate genetically modified animals and cells. One aspect of this invention involves a vector that comprises a sperm cell and one or more polynucleotide molecules bound to a sperm cell through one or more non-liposome based linkers. In a preferred embodiment of this invention, the linker is a protein or polypeptide, preferably sperm specific such as an antibody that binds with the external surface of the sperm cell. In another aspect of the present invention, genetically modified cells or animals are derived from the fertilization of an animal egg cell with the vector described. In one preferred embodiment, genetic modification occurs with the polynucleotide molecule integrating, wholly or partially, into the cell or animal's genome. Another aspect of the present invention includes cells, such as sperm cells or egg cells, and cell lines that are derived from these genetically modified animals or their descendants. In another aspect of the present invention, the genetically modified animals derived from the use of the sperm vector described above possess certain desired characteristics. Examples of these characteristics include faster growth rates, disease or pathogen resistance, high production of certain proteins in milk, and organs suitable for animal to human xenotransplantation.

The invention discloses gene cloning of goat Ran binding protein 9 (RanBP9) and a positioning detection method of the goat RanBP9 in sperm and relates to the technical field of biology. The method includes the steps of firstly, designing cloning primers; secondly, performing RT-PCR amplification on a sequence, and sequencing to obtain a goat RanBP9 gene sequence; thirdly, analyzing the relative expression quantity of RanBP9 mRNA; fourthly, expressing the RanBP9 in tissue; fifthly, using a sperm cell immunofluorescence method to detect the positioning of the RanBP9 in the sperm. The method has the advantages that the goat RanBP9 gene sequence is obtained, expression of the RanBP9 RNA and the RanBP9 in different tissue and positioning of the RanBP9 in the sperm are researched, and a foundation is laid for researching the functions of the RanBP9 in the growth of goat.

Provided are an androgenetic haploid stem cell line, preparation method and use thereof. Specifically, provided are an androgenetic haploid cell and androgenetic blastula. The nucleus of the cell or the blastula only comprises a haploid autosome and a sex chromosome, the sex chromosome being the X chromosome and containing no Y chromosome. The androgenetic haploid cell of the present application can replace a spermatid as a ligand to generate a fertile animal individual, thus facilitating gene manipulation and being capable of transferring genetic information to the offspring.

A method of qualifying sperm cells is provided. The method comprising: (a) staining the sperm cells with acridine orange so as to obtain stained sperm cells; and (b) using a fluorescence activated cell sorter (FACS), gating events simultaneously stained with a green fluorescence intensity higher than 100 nm and red fluorescence intensity in a range selected from 20-1010 nm, the events representing a cell population of the stained sperm cells, wherein a percentage of the cell population is indicative of the quality of the sperm cells.

Disclosed are methods for isolating sperm cells from an aqueous sample and kits for isolating sperm cells from an aqueous sample.

The invention relates to a three-dimensional inducing method for inducing a spermatogonia stem cell to differentiate into a functional sperm cell in vitro, and belongs to the field of bioengineering. On the basis of acquiring the spermatogonia stem cell, a three-dimensional culture system is established through Matrigel, a key factor for inducing differentiation is added, the spermatogonia stem cell is induced to differentiate in vitro to obtain the sperm cell having a fertilization ability, the sperm cell obtained through induction is identified through an immunocytochemical method, the chromosome number of the sperm cell obtained through induction is detected through fluorescence in situ hybridization (FISH), and the fertilization ability and the viability of the sperm cell obtained through induction are detected through round spermatid injection (ROSI). The application provides an excellent model for the molecular mechanism of the human sperm, and in addition, a functional gamete is provided for an azoospermia patient, so that a male infertility patient can have a child of his own.

Herein are disclosed methods for producing female sperm by incorporating a female's chromosomes into a sperm cell via mostly natural spermatogenesis processes. The methods include transplanting (altered) female stem cells or (altered) cloned germ cells into sterilized testes of a male. Diploid female stem / germ cells are altered in two ways—transdifferentiation to facilitate the expression of spermatogenesis factors (for example, using artificial chromosomes) and / or retrodifferentiation to increase pluripotency and imprinting erasures. The altered cells are transplanted into (artificial) male testes to develop into sperm, which are used to fertilize an egg of a second female, or an egg of the original female. Also disclosed herein are methods for producing male eggs by adding an extra X chromosome to an adult male's germ cells, and cultivating the germ cells in vitro.

The invention relates to the field of cell engineering, and concretely provides a method for in vitro culture of testis tissues and induction to generate spermatid. The method comprises the following steps: 1, preparing culture gel pieces: placing 1.2% agarose gel pieces in a 6-orifice plate, adding a medium, immersing overnight, extracting the used medium, adding the fresh medium, and using the agarose gel pieces as culture gel pieces, wherein the medium comprises 90% of MEM alpha and 10% of KSR; 2, taking 5.5-days and 10.5-12.5-days mouse testes, removing external layer albuginea, and cutting to form block tissue samples; 3, placing the tissues samples on the culture gel pieces, putting the 6-orifice plate in a cell culture box, and culturing; and 4, culturing the 5.5-days mouse testis tissues for 22-25d to obtain round sperms, and culturing the 10.5-12.5-days mouse testis tissues for about 27d to obtain elongated sperms.

The invention relates to a preparation method of a shrimp germ cell chromosome. The preparation method comprises the following steps: culturing testis / ovary by colchicine; performing KCl hypotonic treatment and pre-fixing by a Carnoy fixing solution; fixing by an after-fixing solution, wherein the after-fixing solution comprises absolute ethyl alcohol and glacial acetic acid according to the volume ratio of 1:1; dispersing spermatocytes / oocytes to prepare a cell suspension; dyeing by a Giemsa phosphate buffered solution; and performing microscopical examination under a Motic microscope. The preparation method disclosed by the invention has the following advantages that the operation is simple and efficient and chromosome slides can be mass-produced; sufficient chromosome samples can be fast, simply and conveniently obtained, the obtained samples are uniform in dispersion, shallow in background and easy to observe, and the obtained chromosome number is high in accuracy; the simple and easy-to-operate method is provided for preparing shrimp chromosomes; and the non-toxic fixing solutions are adopted, and the harm to a slide making operator can be reduced.

The present invention relates to an antigenic surface structure of sperm cells associated with the Y chromosome, to the molecules, in particular antibodies, directed against this antigenic structure and to a method for characterizing cells carrying only the Y chromosome through the interaction between this antigenic structure and the molecules directed against said structure.

A method and vector system for delivering of a gene into a human stem cell for therapeutic uses is disclosed. The method includes linking a therapeutic gene to human sperm cells through a linker and fertilizing a human oocyte. The resulting zygote may then be cultured and established as human embryonic stem cells, which may later be differentiated into different types of cells for transplantation into the human body.

The invention belongs to the technical field of sperm cell treatment, and specifically relates to a method for isolating and identifying sperm cells of Buffalo testis, including the following steps: obtaining Buffalo testis, digest with collagenase IV, hyaluronidase and DNase I combined enzyme, sieving, filtering, and taking suspension cells after differential adherence; 2) taking the suspend cells for live cell stain; and 3) separating by flow cytometry. Beneficial effect: the technical scheme is an effective method for separating buffalo sperm cells, By digesting bovine testis tissue with two enzymes, so that a single cell suspension is obtain, the interference of somatic cells is removed by the method of continuous differential adherence, according to the fluorescence intensity of non-adherent cells stained by flow cytometry, the DNA content of cells is judged, and the interference of other non-target cells such as diploid spermatogonia and tetraploid primary meningoblasts is effectively digested and removed to achieve the successful sorting of a large number of haploid spermatozoa.

A method for processing forensic samples that include sperm cells from a perpetrator of sexual assault and epithelial cells that are primarily contributed by the victim is provided. The method includes providing a nanofiber filter that is formed of intermingled nanofibers having diameters of about 700 nm or less, selectively digesting the epithelial cells of a forensic sample and separating the sperm cells of the sample from the digested epithelial cells by filtration of the digest mixture through the nanofiber filter, the sperm cells becoming entrapped in the nanofiber filter. The captured sperm cells may then be digested to form a second digest mixture including digested sperm cell DNA. Using the first digest mixture filtrate and the second digest mixture, respectively, DNA samples may be isolated and DNA profiles may be obtained, the DNA profiles being useful for human identification. An apparatus and a kit for processing forensic samples obtained in sexual assault cases are also provided.

The invention relates to a cell nucleus size and shape measuring method. Testicular tissue slices of male animals are analyzed by using a microscopic image analysis system, so that leptotene stage spermatocytes, pachytene stage spermatocytes and spermatids are quickly and automatically classified. The method has huge advantages of quickness, accuracy and human and material resource saving, and provides an important evaluation basis for reproductive system toxicity evaluation of dogs and other male animals in non-clinical safety evaluation.

A method is described for transferring expressible foreign genes into multi-cellular organisms by means of RNA uptake by sperm cells, followed by endogenous reverse transcription of RNA into cDNA. cDNA molecules are transferred to embryos through fertilization, and are transmitted to born animals and their progeny. The cDNA sequences are mosaically distributed in the cells and tissue of the organisms, and are preferentially expressed in target tissues.

The present invention relates to an immunological method that selects spermatozoa having X-chromosome or Y-chromosome. The method of the invention is based on the use of monoclonal antibodies directed against the genus-specific proteins located in the cytoplasm membrane of spermatozoa associated to the action of the classical complement pathway to increase the percentage of one of the gender in the offspring of mammals.

Sperm cell suspensions comprising a motility inhibitor are disclosed. The cells contained in such suspensions tend to have a greater capacity for enduring the various process steps typically associated with the sorting of sperm cells into gender enriched populations, thereby resulting in post-sort compositions with an increased number of viable or motile sperm. Processes for forming such cell suspensions, as well as processes for staining sperm cells, are also disclosed.

The invention provides a dissociation and separation method for shellfish spermary spermatogenic cells, and belongs to the technical field of the separation of ocean shellfish cells. The dissociationand separation method comprises the following steps of: the spermary of a shellfish during a proliferating phase is broken and is digested by a collagenase IV solution of which the mass concentrationis 0.1% and a trypsin solution of which the mass concentration is 0.25% in sequence, and spermary dissociation is carried out to obtain a single-cell suspension; and Percoll solutions of which the volume concentration is independently 10%, 22% and 35% is prepared, and the Percoll solutions are loaded in the same centrifuge tube in a descending order, the single-cell suspension of the shellfish spermatogenic cells is added into the top layer of the centrifuge tube, and centrifugation is carried out to independently collect three layers of cells. By use of a two-step enzymolysis method, spermatogenic cells in spermary tissues can be successfully dissociated into the single-cell suspension, a cell survival rate can be as high as 96.62%, and the cells can be normally subjected to in vitro normal culture. Meanwhile, high-purity spermatogonia and spermatocytes can be separated, an important material is provided for researching a shellfish gamete generating and breeding mechanism, and the dissociation and separation method has a high guidance meaning and a good application value.

A method is provided herein, wherein the method of capturing a sperm deoxyribo nucleic acid (DNA) in a sample, comprises contacting a lysis solution to the sample comprising at least a sperm cell or a sperm cell lysate to lyse the sperm cell. The sperm cell or sperm cell lysate comprises a protamine-DNA complex. The method further comprises applying at least a protamine-specific antibody to the lysed sperm cell, wherein the protamine-specific antibody binds to the protamine-DNA complex of the lysed sperm cell to form an antibody-protamine-DNA complex. The antibody binding is followed by capturing the antibody-protamine-DNA complex; and isolating and detecting the sperm DNA from the captured antibody-protamine-DNA complex.

The invention discloses a method for spreading a mouse spermatocyte chromosome. According to the method, a mouse testicle is grinded, filtered, and centrifugally processed; a NaCl solution is taken asa hypotonic solution; a PBS mixed solution of paraformaldehyde and sodium dodecyl sulfate is taken as a fixing solution to fix mouse spermatocyte on a glass slide; then a mouse SYCP3 monoclonal antibody is taken as a primary antibody; Dylight 549 fluorescence labeled goat-anti-mouse IgG second antibody is taken as a second antibody; a laser confocal microscopy is adopted to observe the spreadingof mouse spermatocyte, and the mitotic phase is recorded at the same time. The method can observe the morphological characteristics of all chromosomes more clearly, and promotes the research and analysis on the meiosis process of spermatocyte and pathogenesis of related diseases.

A method of capturing a sperm deoxyribo nucleic acid (DNA) in a sample is disclosed. The method includes a step of contacting a lysis solution to the sample that includes at least a sperm cell or a sperm cell lysate to lyse the sperm cell. The sperm cell or sperm cell lysate includes a protamine-DNA complex. The method further includes applying at least a protamine-specific antibody to the lysed sperm cell, wherein the protamine-specific antibody binds to the protamine-DNA complex of the lysed sperm cell to form an antibody-protamine-DNA complex. The method further includes capturing the antibody-protamine-DNA complex; and isolating and detecting the sperm DNA from the captured antibody-protamine-DNA complex.

The invention belongs to the technical field of inhibition of hypoxia spermatogenesis reduction, and discloses a detection method for inhibition of hypoxia spermatogenesis reduction of DR5 by VHA. VHAgene knockout mice and C57BL / 6 mice are used as research objects, and a hypoxia spermatogenesis reduction model is constructed through hypoxia induction. A seminiferous tubule seminiferous cell of anexperimental mouse is separated to serve as a research object. Hypoxic in-vitro stimulation is conducted, and the regulating effect and the molecular mechanism of VHA for inhibiting seminiferous cellexpression DR5 are verified through western blot, immunocytochemistry and qPCR technologies. According to the detection method for inhibiting DR5 spermatogenesis reduction under oxygen deficit by VHA, which is provided by the invention, the effect of hypoxic inhibition of VHA in hypoxic spermatogenesis reduction and the molecular mechanism of hypoxic inhibition of VHA in inhibition of DR5 expression are clarified, and a necessary theoretical basis and a new treatment strategy are provided for treatment of hypoxic spermatogenesis reduction.