EP0872219B1 - Anwendung des biofrequenzspektrums in der embryotechnologie bei tieren - Google Patents

Anwendung des biofrequenzspektrums in der embryotechnologie bei tieren Download PDF

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Publication number
EP0872219B1
EP0872219B1 EP95936416A EP95936416A EP0872219B1 EP 0872219 B1 EP0872219 B1 EP 0872219B1 EP 95936416 A EP95936416 A EP 95936416A EP 95936416 A EP95936416 A EP 95936416A EP 0872219 B1 EP0872219 B1 EP 0872219B1
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EP
European Patent Office
Prior art keywords
bio
medium
embryos
vitro
spectrum
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EP95936416A
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English (en)
French (fr)
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EP0872219A4 (de
EP0872219A1 (de
Inventor
Lin Zhou
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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D19/00—Instruments or methods for reproduction or fertilisation
    • A61D19/04—Instruments or methods for reproduction or fertilisation for embryo transplantation

Definitions

  • the present invention relates to biological engineering, particularly to the application of bio-spectrum non-human embryonic engineering.
  • embryonic engineering as used in the following description relates to non-human animal embryos only.
  • the fundamental researches and technical development on animal embryonic engineering have achieved great progress in recent years.
  • the researches on embryonic engineering are moving quickly from the stage of laboratory test to practicalization and commercialization.
  • the animal embryonic engineering mainly includes production of embryos in vitro, cryo-preservation of embryos and micro-manipulation of embryos, etc.
  • the development of the technique for production of embryos in vitro can make full use of animal genetic resources, accelerate improvement of animal gene.
  • the technique can overcome infertility of some rare animals and preserve resources of rare animals.
  • the technique can also supply embryos for production of gene transfer in animals and embryos' sexing. However, the production efficiency and the qualities of embryos produced in vitro are lower. The rate of pregnancy is also lower by embryo transfer.
  • Cryo-preservation of animal embryos is the key technique, which affect whether or not the technique of embryo transfer can be put into practical use.
  • the animal embryos reservoir can be established to facilitate the transportation and exchange of animal breed resources internationally.
  • the cryo-preservation There are two internationally acceptable methods of cryo-preserving embryos. One is the slow freezing method, the other is the cryo-preservation.
  • the survival rate of frozen/thawed embryos produced in vitro is about 65%, and the pregnant rate of the frozen/thawed embryos is decreased 20% than the fresh embryos.
  • Micro-manipulation of embryos consists of sexing of embryos, embryos'cloning including embryo bisection and nuclear transfer, and gene transfer.
  • the aim of sexing of embryos is to produce defined sexual offspring. Cloning of embryos can produce many identical offspring from an outstanding animal embryo, improving the reproduction efficiency and speeding the animal breeding.
  • Gene transfer animals can be produced by microinjection and sperm mediated method. The aim of gene transfer technique is to speed up the growth rate and increase resistance to diseases of animals, improve quality of animal production, and therefore supply many valuable medicines for human beings. However, the efficiency of micro-manipulation is very low because the embryos will definitely be injured.
  • SU-A-1724204 discloses the use of infra-red laser radiation having a particular wavelength during the processing of animal reproduction sperm ;
  • SU-A-731 976 discloses a method for improving fertility of lab rodents by subjecting the females to the irradiation of cm band ;
  • SU-A-1757676 discloses a method of irradiating the chicken embryos with He-Ne laser having a particular wavelength of 632.8 nm ;
  • SU-A-1152583 discloses a method of treatment using pure magnetic field ;
  • SU-A-1 402 343 discloses an animal sperm treatment using pure magnetic field and
  • US 493 0505 discloses a therapeutic method using a laser having a particular wavelength.
  • the invention is based on the following understandings. Very few people have conducted research on the regulation of reproductive and growth abilities of animals with physical method, especially application of physical method to embryos' engineering for resolving technical problems.
  • the inventor recognized that all living matter has chemical and physical characteristics at the same time.
  • the living matter has special physical characteristics such as electric charge in cells, etc.
  • the interactions between the substances in living matter which have electric charges and the electromagnetic field in environment can be produced when electromagnetic field in environment and some main physical characteristics of living matter have the same characteristics. The interactions can influence the molecules, atoms and electrons at the same time to produce significant biologic effects. For example, tissues and cells can grow and develop normally in the chemical and physical environment of a living body.
  • the growth and development of tissues and cells are decreased significantly when they are separated from the living body, although many kinds of chemical protecting materials and nutritive materials are used for improving the culture conditions.
  • the inventor further recognized that the growth ability of tissues and cells in culture condition will be improved by applying a simulated bio-spectrum which is a weak electromagnetic field. So, it has significant meanings to apply the simulated bio-spectrum to embryos' engineering. The bio-spectrum will help improve the reproductive ability, growth speed and resistance to diseases of the animals.
  • the object of the invention is to induce irradiation bio-effects by applying the bio-spectrum to animal embryos engineering and animals, which includes:
  • the object of the present invention is realized by the following technical solutions.
  • the oocytes are collected from donor cows by ordinary methods, and than put in common medium or special medium for maturation. During maturation, oocytes are irradiated with bio-spectrum device (model WS-101D) for 15 minutes (at weak level). The temperature should be kept at 38-40 °C by adjusting the distance between embryo container and irradiation device.
  • bio-spectrum device model WS-101D
  • the special medium is made according to the procedure of Brackett et al.
  • the receipt of the special medium is as follows: components g/l NaCl 6.55 KCl 0.30 CaCl 2 ⁇ 2H 2 O 0.33 NaH 2 PO 4 ⁇ H 2 O 0.11 MgCl 2 ⁇ 6H 2 O 0.11 NaHCO 3 3.10 Glucose 2.25 Bovine Serum Albumin 3.00 Sodium Pyruvate (or Pyruvate) 0.14(or 0.11) Na-Penicillin 0.031(50 IU/ml)
  • the medium needs to be sterilized by filtration after the components have dissolved completely in 1000 ml water. This medium is required to equilibrate in the incubator at 38 °C in 59% CO 2 in the air.
  • the osmolality of this medium is about 300 mOsm/kg H 2 O.
  • the special medium is made by addition of 84 mg NaHCO 3 to 100 ml so-prepared sulotion.
  • the m-HIS is made by addition of 34 mg NaCI to 10 ml medium.
  • the fresh semen is firstly treated following the procedure of Bracktt et al. Semen is pre-diluted with special medium (350g). After centrifugation, the sperm pellet is re-suspended with m-HIS medium and put in water bath of 38 °C for 15 minutes. After another centrifugation, the sperm pellet is re-suspended with special medium in a tube for irradiation with bio-spectrum device at weak level for 10 minutes. During irradiation, the temperature of medium should be kept at 38-40 °C by adjusting the distance between the tube and irradiation device.
  • the in-vitro matured oocytes and in-vitro capacitated sperm are put in the special medium in a tube for irradiation with bio-spectrum device for 20 minutes.
  • the tube is put in CO 2 incubator for one hour culture at 38 °C.
  • the tube is irradiated with bio-spectrum again for 18 minutes and again put in incubator for 5-hour culture.
  • weak level is used and the temperature of medium is kept at 38-40 °C by adjusting the distance between the tube and irradiation device.
  • the in-vitro fertilized eggs are put in the special medium for irradiation with BIO-SPECTRUM for 25 minutes. Weak level is used. During irradiation the temperature of medium is kept at 38-40 °C by adjusting the distance between the medium container and irradiation device.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Reproductive Health (AREA)
  • Transplantation (AREA)
  • Zoology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Claims (2)

  1. Verfahren zur Anwendung einer Bestrahlung mit einem Bio-Frequenzspektrum in der Embryonen-Technologie nicht-menschlicher, tierischer Embryonen, wobei das genannte Bio-Frequenzspektrum über Wellenlängen im Bereich zwischen 0,2 µm und 10 cm verfügt und wobei die genannte Embryonen-Technologie nicht-menschlicher, tierischer Embryonen umfasst
    (1) in-vitro-Reifung gewonnener Oozyten,
    (2) Kapazitation von Spermatozoen,
    (3) in-vitro-Kultur von Embryonen,
    wobei während der in-vitro-Reifung Oozyten, die in einem Standardmedium oder in definiertem Medium kultiviert wurden, mit einem Bio-Spektrum-Generator 3 bis 20 Minuten lang bestrahlt werden, wobei die Temperatur des Mediums während der Bestrahlung auf nicht mehr als 40°C gehalten wird; während der Kapazitation wird mit einem Standardmedium oder definiertem Medium verdünntes Semen mit dem Bio-Spektrum-Generator 3 bis 20 Minuten lang bestrahlt, wobei die Temperatur des Mediums während der Bestrahlung nicht mehr als 40°C beträgt;
    während der in-vitro-Fertilisation werden sowohl Oozyten, als auch in-vitro-kapazitierte Spermatozoen zusammen in einem Standardmedium oder definiertes Medium enthaltenden Reagenzglas kultiviert und mit dem Bio-Spektrum-Generator einbis dreimal jeweils 3 bis 25 Minuten lang bestrahlt, wobei die Temperatur des Mediums während der Bestrahlung nicht mehr als 40°C beträgt;
    während der in-vitro-Kultur von Embryonen werden potentielle Zygoten in ein Standardmedium oder in definiertes Medium übertragen und mit dem Bio-Spektrum-Generator 3 bis 20 Minuten lang bestrahlt, wobei die Temperatur des Mediums während dieser Zeit nicht mehr als 40°C beträgt.
  2. Verfahren nach Patentanspruch 1, in dem die Bestandteile des genannten definierten Mediums 6,55 g/l NaCI, 0,30 g/l KCI, 0,33 g/l CaCl2·2H2O, 0,11 g/l NaH2PO4 · H2O, 0,11 g/l MgCl2·6H2O, 3,10 g/l NaHCO3, 2,5 g/l Glukose, 0,14 g/l Natriumpyruvat (0,11 g/l Pyruvat), 3,00 g/l Albumin aus Rinderserum, 0,031 g/l (50 IE/ml) Natriumsalz von Penicillin sind und 84 mg NaHCO3 100ml des genannten definierten Mediums zugesetzt werden.
EP95936416A 1994-11-24 1995-11-02 Anwendung des biofrequenzspektrums in der embryotechnologie bei tieren Expired - Lifetime EP0872219B1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CN94118306 1994-11-24
CN 94118306 CN1068195C (zh) 1994-11-24 1994-11-24 生物频谱在胚胎工程中的应用
PCT/CN1995/000087 WO1996015732A1 (fr) 1994-11-24 1995-11-02 Utilisation du spectre de frequences biologiques dans l'embryogenese animale

Publications (3)

Publication Number Publication Date
EP0872219A1 EP0872219A1 (de) 1998-10-21
EP0872219A4 EP0872219A4 (de) 1999-02-03
EP0872219B1 true EP0872219B1 (de) 2002-09-18

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EP95936416A Expired - Lifetime EP0872219B1 (de) 1994-11-24 1995-11-02 Anwendung des biofrequenzspektrums in der embryotechnologie bei tieren

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EP (1) EP0872219B1 (de)
CN (1) CN1068195C (de)
AU (1) AU704690B2 (de)
CA (1) CA2207013C (de)
DE (1) DE69528295T2 (de)
WO (1) WO1996015732A1 (de)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IL137366A0 (en) * 2000-07-18 2001-07-24 Shladot Metal Works Ltd A method for increasing the fertilizing capability of sperm cells
KR100516881B1 (ko) * 2002-12-13 2005-09-27 (주)아비코아생명공학연구소 Ｘ선 조사를 이용한 체세포 복제수정란의 제조를 위한수핵난자 핵 유전 물질의 불활화 방법
CN102250832B (zh) * 2011-05-30 2012-08-15 中国农业大学 一种促进冷冻胚胎解冻后体外发育的培养液

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU731976A1 (ru) * 1975-04-18 1980-05-05 Ленинградский Научно-Исследовательский Институт Радиационной Гигиены Способ повышени плодовитости мелких грызунов
SU1152583A1 (ru) * 1982-03-01 1985-04-30 Кубанский Ордена Трудового Красного Знамени Сельскохозяйственный Институт Способ осеменени свиноматок
EP0263192A1 (de) * 1986-10-03 1988-04-13 HENKEL CORPORATION (a Delaware corp.) Dicyanethenyl-Fettverbindungen und deren Derivate
SU1402343A1 (ru) * 1985-06-04 1988-06-15 Кубанский сельскохозяйственный институт Способ обработки спермы животных и устройство дл его осуществлени
SU1724204A1 (ru) * 1989-10-09 1992-04-07 Гродненский государственный медицинский институт Способ обработки спермы хр ков
SU1757676A1 (ru) * 1989-11-27 1992-08-30 Горский Сельскохозяйственный Институт Способ стимул ции образовани специфического иммунитета у кур к болезни Ньюкасла

Also Published As

Publication number Publication date
EP0872219A4 (de) 1999-02-03
WO1996015732A1 (fr) 1996-05-30
DE69528295D1 (de) 2002-10-24
CN1068195C (zh) 2001-07-11
CA2207013C (en) 2006-07-11
CA2207013A1 (en) 1996-05-30
DE69528295T2 (de) 2003-05-08
CN1123131A (zh) 1996-05-29
EP0872219A1 (de) 1998-10-21
AU704690B2 (en) 1999-04-29
AU3838695A (en) 1996-06-17

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