WO1984001265A1 - Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y - Google Patents

Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y Download PDF

Info

Publication number
WO1984001265A1
WO1984001265A1 PCT/US1982/001478 US8201478W WO8401265A1 WO 1984001265 A1 WO1984001265 A1 WO 1984001265A1 US 8201478 W US8201478 W US 8201478W WO 8401265 A1 WO8401265 A1 WO 8401265A1
Authority
WO
WIPO (PCT)
Prior art keywords
semen
sperm
mammal
quinicrine
dye
Prior art date
Application number
PCT/US1982/001478
Other languages
English (en)
Inventor
Edwin Adair
Original Assignee
Genetic Engineering Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Genetic Engineering Inc filed Critical Genetic Engineering Inc
Priority to EP19830900092 priority Critical patent/EP0120854A4/fr
Priority to PCT/US1982/001478 priority patent/WO1984001265A1/fr
Publication of WO1984001265A1 publication Critical patent/WO1984001265A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/52—Sperm; Prostate; Seminal fluid; Leydig cells of testes

Definitions

  • This invention relates to an improved method of preserving collected mammalian semen and separating the sperm contained therein into X and Y components so that a female of the species may be artificially inseminated with separated sperm to produce offspring of a desi red and predetermined sex .
  • Th is invention is particularly useful for preserving cattle semen and separating it so that sperm bearing only X ch romosomes can be sepa rated and used to produce dairy herds and sperm bearing only Y ch romosomes can be used to produce beef herds .
  • This invention also has usage in breeding horses and other animais .
  • Some physical cha racteristics of sperm that are of potential use in separating sperm cells are: higher density or packing of the Y ch romosome, greater DNA content of the X ch romosome, slightly larger size and heavier weight of sperm contain i ng the X ch romosome; generally h igher velocity of movement of the Y ch romosome containing sperm; and an appa rent dense negative electrical su rface charge on the X ch romosome .
  • the methods of separation of sperm utilizing these physical properties have, for the most part, been on ly partially successfu l , with separation percentages ranging f rom 60° o to 80%.
  • SUBSTITUTE SHEET very great need for a method of sperm separation which is virtually 100% accu rate and which can be done very rapidly. Furthermore, in order to accomplish separation , it is necessary to maintain or get the semen which has been collected in a fluid non- coagulated condition before the separation of the sperm can be accomplished .
  • a method of separating viable sperms cells according to whether they carry an X chromosome or a Y chromosome is described.
  • the separated batches of cells are suitable for use in artificial insemination of humans, cows and other livestock animais so that offspring of predetermined sex are obtained.
  • the method of separation depends on basic differences in X and Y sperm cells. For example, each X sperm has a greater DNA content then a corresponding Y sperm since the X chromosome has a greater DNA content than a comparable Y chromosome. Hence, it is postulated that the X sperm should incorporate greater absolute amounts of certain dyes and other labels then Y sperm. On the other hand, it is believed that the Y chromosome of Y sperm is more dense or tightly packed than a corresponding X chromosome.
  • Semen which contains sperm is collected from a mammal and treated by adding an anti-coagulant, such as mammal saliva, to prevent coagulation and hasten liquefaction. Any anticoagulant that does not affect viability will work. In particular, human saliva has been found very satisfactory for this purpose and is readily available.
  • the semen is then diluted with a diluting solution and stored in a water bath at substantially the body temperature of the mammal or the sperm cells are centrifuged out of the semen fluid and diluted in a suitable saline solution.
  • a cell membrane diffusion material such as sperm-free human semen
  • sperm-free human semen can be added to the mammalian semen so that a fluorescent dye can penetrate the cell membrane of the sperm cell and become localized around the chromosomes therein. The dye will penetrate the cell membranes of the sperm cell of primates and vole semen without prior preparation.
  • SUBSTITUTE SHEET The selection of dye depends on the method of detection used to select the X and Y sperm. If a scanning procedu re is used wherein any bright spots of fluorescent dye would be detected, such as that which it is postulated characterize the Y chromosome because of its higher density than an ideal dye is quinicrine or quinicrine mustard .
  • a more suitable dye has been found to be acridine, acridtne orange and derivatives thereof such as ethidium, bromide, mith ramycin or any combination thereof .
  • Another suitable dye is DAPI (4, 6-diamidino-2-phenylindole) .
  • the semen is put into a detection chamber.
  • the semen is flowed through the chamber so that ideally only one sperm ceil at a time is irradiated with ultraviolet light.
  • the light causes the dye to fluorescence.
  • a detection devise in the chamber senses the degree of fluorescence and provides an output signal in response to the sensed cell which varies according to the degree of fluorescence.
  • the stream then passes through a fluidic amplifier and the stream is switched in response to each sensed cel l in the detection chamber to direct the sensed cell to one of two outlet ports .
  • the detection device is programmed to scanning the cell for bright spots the Y sperm will trigger the system presumably because of the higher density of the dye localized around the denser Y ch romosome. If instead the greater absolute degree of fluorescence is being detected , the X sperm will trigger the system presumably because the X sperm contains more DNA; and, hence more dye is localized a round it. I n any way, the X and Y ch romosomes can be effectively separated into separate containers for use in artificial insemination of female animals in order to produce the desi red sexed offspring .
  • Semen is first collected from a mammal in any manner which is known to one skilled in the art. For example, with cattle and horses, teaser animals or electrical stimulation may be used. The various methods of collecting semen from animals is explained in conventional vetenarian reference texts.
  • the collected semen is placed in a clean, sterile container and may be treated with an enzyme solution to hasten liquefaction.
  • Cattle semen for example is highly viscous and will not flow in a stream as necessary for separation unless an anti-coagulant is added and a dilutant is also added.
  • one of the best enzyme solutions and one of the most physiologically compatible is mammalian saliva, and particularly human saliva.
  • about 1 ml of human saliva is added to each quantity of about 2.0 ml to about 3.5 ml of semen. If the quantity of human saliva added is significantly less than 1 ml, the reaction will be too slow. However, an amount larger than 1 ml will have no adverse effect, but is unnecessary.
  • semen diluting solution contains egg yolk, glycerol, glucose and citrate. Preparation of these solutions is set forth in Fertility and Sterility by Berman and Sasada, published in 1966.
  • One such anti-coagulant is alpha-amylase.
  • the semen is then stored in a water bath at approximately the body temperature of the animal. This is between about 35°C and about 38.5°C, but is usually about 37 C. If the temperature is substantially above or below this range, sperm mobility will decrease and the survival rate of the sperm will decrease below acceptable limits. When it is time to selectively identify and separate the X and
  • a cell membrane diffusion material must be added to the collected semen so that the dye can penetrate the cell. The dye will penetrate in primate and vole semen without further treatment.
  • DMSO dimethylsuifoxide
  • disulfomethoxide SUBSTITUTE ET membrane diffusion material which has been found satisfactory. It has been found that the addition of about 0.05 ml of DMSO to a quantity of about 2.0 ml to about 3.5 ml of semen provides sufficient penetration of the cell membrane by the dye.
  • sperm-free human semen Another cell membrane diffusion material which has been found quite satisfactory is sperm-free human semen .
  • the sperm - free human semen is added in a quantity of about 0. 5 ml human semen to a quantity of about 2.0 mi to about 3.5 mi of mammal semen . If either DMSO or human semen is added in quantities substantially below the indicated ranges , the diffusion of the dye will be very slow whereas if the amounts added are substantially above the indicated ranges, the DMSO and human semen will have a toxic effect and kill substantial numbers of sperm .
  • the dyes which has been found most satisfactory are taken from the group consisting of quinicrine, quinicrine hydrochloride, quinicrine dihydrochloride and quinicrine mustard .
  • quinicrine hydrochloride or quinicrine dihydrochloride is used , it is diluted in a sol ution of ion-free distilled water to a strength in the range of about 0.05° o to about 5.0° .
  • the most satisfactory range has been found to be between about 0.5° o and about 1 .0° o in sol ution .
  • quinicrine mustard When quinicrine mustard is used , it is diluted with ion- free distilled water to a solution having a strength in the range of about 0.001°o to about 0.01° o whereas the optimum range is about 0.005°o solution . If the solutions are substantially below the indicated range in concentration , the dye will ta ke too long to stain , whereas if the solutions are substantially above the indicated ranges in concentration , the dye will have a toxic effect and kill substantial numbers of the sperm . The dye solution is put in contact for a period of at least several minutes , but usually for a period of at least 15 minutes to assu re good dye penetration . Confirmation of staining is obtained by separating a small part of the specimen and checking for staining under a fluorescent microscope.
  • the difference in fluorescence of X and Y chromosome bearing cells entrained in a stream of diluted semen is detected and the resultant different signals which are produced as a result of this detection are used to control a fluidic amplifier located downstream to switch the X sperm to one outlet port and the Y sperm to another outlet port so they can be separated and collected.
  • the semen is fed in a very narrow stream so that the sperm cells in the semen move essentially single file past a laser light beam which is provided with an ultraviolet filter.
  • the laser projects a very narrow beam in which a pattern of illumination of the beam, when it strikes the sperm cell, appears as a thin line of light transverse to the stream of cells.
  • Electrical photoresponsive pick-up elements are arranged around the outside of the detection chamber through which the stream passes to detect any scattering of light due to the ultraviolet light striking a fluorescent Y chromosome.
  • the excitation wavelength of the laser beam is in the range of 457 nm to 488 nm.
  • the emission wavelength from the fluorescent Y sperm will be around 351 nm.
  • the fluid amplifier includes a transducer which is responsive to any scattering of light due to fluorescence of a Y sperm which it is postulated will create a turbulance in the stream to cause a "wall attachment effect" as is well known in fluidics.
  • any X chromosome it is postulated will pass along the stream uninterrupted and out a first outlet port whereas the Y chromosomes will be diverted due to reaction of the transducer to the scattered light causing them to be diverted and pass through a second outlet port.
  • the semen can be separated into X and Y sperm and separately collected for

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Cell Biology (AREA)
  • Reproductive Health (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Procédé de sexualisation de cellules spermatiques viables en utilisant un cytomètre en combinaison avec un marquage sélectif de cellules spermatiques avec un agent de coloration fluorescent. Par excitation de cellules portant des chromosomes X et Y marqués sélectivement, un système de détection est activé de telle sorte que des cellules spermatiques X et Y sont canalisées dans deux orifices séparés de récupération.
PCT/US1982/001478 1982-10-05 1982-10-05 Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y WO1984001265A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP19830900092 EP0120854A4 (fr) 1982-10-05 1982-10-05 Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y.
PCT/US1982/001478 WO1984001265A1 (fr) 1982-10-05 1982-10-05 Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US1982/001478 WO1984001265A1 (fr) 1982-10-05 1982-10-05 Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y

Publications (1)

Publication Number Publication Date
WO1984001265A1 true WO1984001265A1 (fr) 1984-04-12

Family

ID=22168295

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1982/001478 WO1984001265A1 (fr) 1982-10-05 1982-10-05 Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y

Country Status (2)

Country Link
EP (1) EP0120854A4 (fr)
WO (1) WO1984001265A1 (fr)

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0235046A1 (fr) * 1986-02-28 1987-09-02 INSTITUT NATIONAL DE LA RECHERCHE AGRONOMIQUE, Etablissement public dit: Sondes d'ADN spécifique du génome mâle des ruminants, leur préparation et utilisation
WO1990004404A1 (fr) * 1988-10-25 1990-05-03 The Victoria University Of Manchester Procede gynecologique
US5021244A (en) * 1988-12-06 1991-06-04 Cytogam, Inc. Sex-associated membrane antibodies and their use for increasing the probability that offspring will be of a desired sex
EP0475936A1 (fr) * 1989-05-12 1992-03-25 Cytogam Inc Proteines membranaires associees au sexe et procedes pour accroitre la probabilite que la progeniture sera d'un sexe desire.
US5346990A (en) * 1987-04-08 1994-09-13 Cytogam, Inc. Sex-associated membrane proteins and methods for increasing the probability that offspring will be of a desired sex
EP0658345A1 (fr) * 1989-05-12 1995-06-21 Cytogam, Inc. Protéines membranaires associées au sexe et procédés pour accroître la probabilité que la progéniture sera d'un sexe désiré
EP0471758B1 (fr) * 1989-05-10 1996-09-11 THE UNITED STATES OF AMERICA as represented by the Secretary UNITED STATES DEPARTMENT OF COMMERCE Procede de preselection du sexe des progenitures
US6153373A (en) * 1997-07-01 2000-11-28 Vicam, L.P. Method for sex determination of mammalian offspring
WO2005042721A2 (fr) * 2003-10-31 2005-05-12 Abs Global, Inc. Procede de modification de la proportion males-femelles de la descendance de mammiferes par la manipulation des spermatozoides
US7713687B2 (en) 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
US7723116B2 (en) 2003-05-15 2010-05-25 Xy, Inc. Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm
US7758811B2 (en) 2003-03-28 2010-07-20 Inguran, Llc System for analyzing particles using multiple flow cytometry units
US7820425B2 (en) 1999-11-24 2010-10-26 Xy, Llc Method of cryopreserving selected sperm cells
US7833147B2 (en) 2004-07-22 2010-11-16 Inguran, LLC. Process for enriching a population of sperm cells
US7838210B2 (en) 2004-03-29 2010-11-23 Inguran, LLC. Sperm suspensions for sorting into X or Y chromosome-bearing enriched populations
US7855078B2 (en) 2002-08-15 2010-12-21 Xy, Llc High resolution flow cytometer
US7929137B2 (en) 1997-01-31 2011-04-19 Xy, Llc Optical apparatus
US8137967B2 (en) 2000-11-29 2012-03-20 Xy, Llc In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations
US8486618B2 (en) 2002-08-01 2013-07-16 Xy, Llc Heterogeneous inseminate system
US8497063B2 (en) 2002-08-01 2013-07-30 Xy, Llc Sex selected equine embryo production system
US9145590B2 (en) 2000-05-09 2015-09-29 Xy, Llc Methods and apparatus for high purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa
US9365822B2 (en) 1997-12-31 2016-06-14 Xy, Llc System and method for sorting cells
US11230695B2 (en) 2002-09-13 2022-01-25 Xy, Llc Sperm cell processing and preservation systems
CN114885939A (zh) * 2022-04-20 2022-08-12 新疆泰昆集团有限责任公司 一种鸡精液稀释液及其制备方法和应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3654084B2 (ja) 1999-09-27 2005-06-02 ヤマハ株式会社 波形生成方法及び装置

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3185623A (en) * 1960-05-31 1965-05-25 Smith Fred Preservation of animal semen
US3207910A (en) * 1959-03-12 1965-09-21 Int Standard Electric Corp Photosensitive arrangement for scanning fluorescing identifications
US3305089A (en) * 1965-08-13 1967-02-21 Gunsons Sortex Ltd Apparatus for sorting fluorescent articles
US3586859A (en) * 1970-02-16 1971-06-22 Irwin J Katz Fluorescent cell viability counter
US3816249A (en) * 1970-11-23 1974-06-11 B Bhattacharya Universal medium and method for extending the useful life of semen in vitro
US4092229A (en) * 1975-12-17 1978-05-30 Bhattacharya Bhairab C Thermal convection counter streaming sedimentation and forced convection galvanization method for controlling the sex of mammalian offspring

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
BE677791A (fr) * 1965-03-29 1966-09-14
DE2154259A1 (de) * 1971-10-30 1973-05-10 Schaumann Fa H Wilhelm Verfahren zum konservieren von ebersperma bei 13 bis 17 grad celsius
US3791517A (en) * 1973-03-05 1974-02-12 Bio Physics Systems Inc Digital fluidic amplifier particle sorter
US4362246A (en) * 1980-07-14 1982-12-07 Adair Edwin Lloyd Method of treating collected mammal semen and separating sperm into X Y components

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3207910A (en) * 1959-03-12 1965-09-21 Int Standard Electric Corp Photosensitive arrangement for scanning fluorescing identifications
US3185623A (en) * 1960-05-31 1965-05-25 Smith Fred Preservation of animal semen
US3305089A (en) * 1965-08-13 1967-02-21 Gunsons Sortex Ltd Apparatus for sorting fluorescent articles
US3586859A (en) * 1970-02-16 1971-06-22 Irwin J Katz Fluorescent cell viability counter
US3816249A (en) * 1970-11-23 1974-06-11 B Bhattacharya Universal medium and method for extending the useful life of semen in vitro
US4092229A (en) * 1975-12-17 1978-05-30 Bhattacharya Bhairab C Thermal convection counter streaming sedimentation and forced convection galvanization method for controlling the sex of mammalian offspring

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0120854A4 *

Cited By (58)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2603701A2 (fr) * 1986-02-28 1988-03-11 Agronomique Inst Nat Rech Applications des sondes moleculaires d'adn specifique du genome male de mammiferes, notamment du genre bos, au controle de la presence du chromosome y dans une population de spermatozoides et a la separation des spermatozoides en deux populations de spermatozoides porteurs respectivement du chromosome y et du chromosome x pour leur utilisation pour l'insemination artificielle
EP0235046A1 (fr) * 1986-02-28 1987-09-02 INSTITUT NATIONAL DE LA RECHERCHE AGRONOMIQUE, Etablissement public dit: Sondes d'ADN spécifique du génome mâle des ruminants, leur préparation et utilisation
US5439362A (en) * 1987-04-08 1995-08-08 Cytogam, Inc. Sex-associated membrane proteins and methods for increasing the probability that offspring will be of a desired sex
US5660997A (en) * 1987-04-08 1997-08-26 Cytogam, Inc. Methods for determining antibodies specific for sex associated sperm membrane proteins
US5346990A (en) * 1987-04-08 1994-09-13 Cytogam, Inc. Sex-associated membrane proteins and methods for increasing the probability that offspring will be of a desired sex
WO1990004404A1 (fr) * 1988-10-25 1990-05-03 The Victoria University Of Manchester Procede gynecologique
EP0372702A1 (fr) * 1988-10-25 1990-06-13 The Victoria University Of Manchester Procédé gynécologique
US5021244A (en) * 1988-12-06 1991-06-04 Cytogam, Inc. Sex-associated membrane antibodies and their use for increasing the probability that offspring will be of a desired sex
EP0471758B1 (fr) * 1989-05-10 1996-09-11 THE UNITED STATES OF AMERICA as represented by the Secretary UNITED STATES DEPARTMENT OF COMMERCE Procede de preselection du sexe des progenitures
EP0475936A1 (fr) * 1989-05-12 1992-03-25 Cytogam Inc Proteines membranaires associees au sexe et procedes pour accroitre la probabilite que la progeniture sera d'un sexe desire.
EP0658345A1 (fr) * 1989-05-12 1995-06-21 Cytogam, Inc. Protéines membranaires associées au sexe et procédés pour accroître la probabilité que la progéniture sera d'un sexe désiré
EP0475936A4 (en) * 1989-05-12 1992-05-06 Cytogam, Inc. Sex-associated membrane proteins and methods for increasing the probability that offspring will be of a desired sex
US8975035B2 (en) 1997-01-31 2015-03-10 Xy, Llc Method of analyzing cells
US8553226B2 (en) 1997-01-31 2013-10-08 Xy, Llc Optical apparatus
US7929137B2 (en) 1997-01-31 2011-04-19 Xy, Llc Optical apparatus
US6489092B1 (en) 1997-07-01 2002-12-03 Vicam, L.P. Method for sex determination of mammalian offspring
US6153373A (en) * 1997-07-01 2000-11-28 Vicam, L.P. Method for sex determination of mammalian offspring
US9365822B2 (en) 1997-12-31 2016-06-14 Xy, Llc System and method for sorting cells
US9422523B2 (en) 1997-12-31 2016-08-23 Xy, Llc System and method for sorting cells
US7820425B2 (en) 1999-11-24 2010-10-26 Xy, Llc Method of cryopreserving selected sperm cells
US10208345B2 (en) 2000-05-09 2019-02-19 Xy, Llc Method for producing high purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa
US9145590B2 (en) 2000-05-09 2015-09-29 Xy, Llc Methods and apparatus for high purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa
US7713687B2 (en) 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
US7771921B2 (en) 2000-11-29 2010-08-10 Xy, Llc Separation systems of frozen-thawed spermatozoa into X-chromosome bearing and Y-chromosome bearing populations
US9879221B2 (en) 2000-11-29 2018-01-30 Xy, Llc Method of in-vitro fertilization with spermatozoa separated into X-chromosome and Y-chromosome bearing populations
US8652769B2 (en) 2000-11-29 2014-02-18 Xy, Llc Methods for separating frozen-thawed spermatozoa into X-chromosome bearing and Y-chromosome bearing populations
US8137967B2 (en) 2000-11-29 2012-03-20 Xy, Llc In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations
US8486618B2 (en) 2002-08-01 2013-07-16 Xy, Llc Heterogeneous inseminate system
US8497063B2 (en) 2002-08-01 2013-07-30 Xy, Llc Sex selected equine embryo production system
US7855078B2 (en) 2002-08-15 2010-12-21 Xy, Llc High resolution flow cytometer
US11230695B2 (en) 2002-09-13 2022-01-25 Xy, Llc Sperm cell processing and preservation systems
US11261424B2 (en) 2002-09-13 2022-03-01 Xy, Llc Sperm cell processing systems
US8709825B2 (en) 2003-03-28 2014-04-29 Inguran, Llc Flow cytometer method and apparatus
US7943384B2 (en) 2003-03-28 2011-05-17 Inguran Llc Apparatus and methods for sorting particles
US8241914B2 (en) 2003-03-28 2012-08-14 Inguran Llc EPI-damage apparatus and methods for sorting particles
US8206988B2 (en) 2003-03-28 2012-06-26 Inguran Llc Method and apparatus for orienting sperm in a fluid stream
US8198093B2 (en) 2003-03-28 2012-06-12 Inguran Llc Methods for sorting particles
US8198092B2 (en) 2003-03-28 2012-06-12 Inguran, Llc Digital sampling apparatus and methods for sorting particles
EP2308417A3 (fr) * 2003-03-28 2011-08-03 Inguran, LLC Appareil et procédés pour fournir du sperme d'animaux triés selon le sexe
US8664006B2 (en) 2003-03-28 2014-03-04 Inguran, Llc Flow cytometer apparatus and method
US8709817B2 (en) 2003-03-28 2014-04-29 Inguran, Llc Systems and methods for sorting particles
US10100278B2 (en) 2003-03-28 2018-10-16 Inguran, Llc Multi-channel system and methods for sorting particles
US8748183B2 (en) 2003-03-28 2014-06-10 Inguran, Llc Method and apparatus for calibrating a flow cytometer
US11718826B2 (en) 2003-03-28 2023-08-08 Inguran, Llc System and method for sorting particles
US9040304B2 (en) 2003-03-28 2015-05-26 Inguran, Llc Multi-channel system and methods for sorting particles
US11104880B2 (en) 2003-03-28 2021-08-31 Inguran, Llc Photo-damage system for sorting particles
US8206987B2 (en) 2003-03-28 2012-06-26 Inguran Llc Photo-damage method for sorting particles
US9377390B2 (en) 2003-03-28 2016-06-28 Inguran, Llc Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm
US7799569B2 (en) 2003-03-28 2010-09-21 Inguran, Llc Process for evaluating staining conditions of cells for sorting
US7758811B2 (en) 2003-03-28 2010-07-20 Inguran, Llc System for analyzing particles using multiple flow cytometry units
US7723116B2 (en) 2003-05-15 2010-05-25 Xy, Inc. Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm
WO2005042721A3 (fr) * 2003-10-31 2006-09-28 Abs Global Inc Procede de modification de la proportion males-femelles de la descendance de mammiferes par la manipulation des spermatozoides
WO2005042721A2 (fr) * 2003-10-31 2005-05-12 Abs Global, Inc. Procede de modification de la proportion males-femelles de la descendance de mammiferes par la manipulation des spermatozoides
US7838210B2 (en) 2004-03-29 2010-11-23 Inguran, LLC. Sperm suspensions for sorting into X or Y chromosome-bearing enriched populations
US7892725B2 (en) 2004-03-29 2011-02-22 Inguran, Llc Process for storing a sperm dispersion
US7833147B2 (en) 2004-07-22 2010-11-16 Inguran, LLC. Process for enriching a population of sperm cells
CN114885939A (zh) * 2022-04-20 2022-08-12 新疆泰昆集团有限责任公司 一种鸡精液稀释液及其制备方法和应用
CN114885939B (zh) * 2022-04-20 2023-02-24 新疆泰昆集团有限责任公司 一种鸡精液稀释液及其制备方法和应用

Also Published As

Publication number Publication date
EP0120854A1 (fr) 1984-10-10
EP0120854A4 (fr) 1985-06-10

Similar Documents

Publication Publication Date Title
US4362246A (en) Method of treating collected mammal semen and separating sperm into X Y components
WO1984001265A1 (fr) Procede de traitement de sperme recupere chez un mammifere et de separation du sperme en composants x et y
Johnson Sex preselection by flow cytometric separation of X and Y chromosome-bearing sperm based on DNA difference: a review
JP2552582B2 (ja) 子の性の前選択の方法
US7833147B2 (en) Process for enriching a population of sperm cells
US20080108046A1 (en) Method of sorting cells
AU2013325222B2 (en) Methods of processing sperm for sex sorting
GB2145112A (en) Sorting living spermatozoa
WO2001051612A1 (fr) Procede de production d'embryons non humains presentant une valeur genetique elevee et un sexe predetermine
CA1250808A (fr) Sexage du liquide seminal
Canfield et al. Normal haematological and biochemical values for the swamp buffalo (Bubalus bubalis) in Australia
Owens et al. Use of an in vitro biotinylation technique for determination of posttransfusion survival of fresh and stored autologous red blood cells in Thoroughbreds
Howes et al. A search for sex-specific antigens on bovine spermatozoa using immunological and biochemical techniques to compare the protein profiles of X and Y chromosome-bearing sperm populations separated by fluorescence-activated cell sorting
Jain et al. Efficient production of sexed semen by flow cytometry: a review
Dybas Cellular defense reactions of Phascolosoma agassizii, a sipunculan worm: Phagocytosis by granulocytes
Nagy et al. Association between the strength of serologic recognition of bovine leukosis virus and lymphocyte count in bovine leukosis virus-infected cows
Du Pasquier et al. Transplantation of nuclei from lymphocytes of adult frogs into enucleated eggs: special focus on technical parameters
Osama et al. Effect of Rabbit Semen Filtration Methods on Conception Rates and Altering Sex Ratio in Artificially Inseminated Rabbits.
Dadah et al. Trypanosomosis among sheep and goats at slaughter in Jos abattoir, Nigeria
Bhojani et al. Clinico-therapeutic studies on Theileria equi infection in horses
Suarez et al. Binding of a marker for immunoglobulins to the surface of rabbit testicular, epididymal, and ejaculated spermatozoa
Andamin et al. SEROPREVALENCE AND HAEMATOLOGICAL STUDIES OF SOME WILD AND SEMI-DOMESTICATED BIRDS NATURALLY INFECTED WITH AVIAN POX VIRUS IN ZARIA, NGERIA
Kimeto Pathology and pathogenesis of Trypanosoma vivax infection in cattle
LEROUX et al. veterinaryrecord. bmj. com
Keeler Flow microfluorometric analysis of living spermatozoa

Legal Events

Date Code Title Description
AK Designated states

Designated state(s): JP

AL Designated countries for regional patents

Designated state(s): CH DE FR GB

WWE Wipo information: entry into national phase

Ref document number: 1983900092

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1983900092

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1983900092

Country of ref document: EP