WO2004033729A1 - Method of preserving biological materials - Google Patents

Method of preserving biological materials Download PDF

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Publication number
WO2004033729A1
WO2004033729A1 PCT/RU2003/000070 RU0300070W WO2004033729A1 WO 2004033729 A1 WO2004033729 A1 WO 2004033729A1 RU 0300070 W RU0300070 W RU 0300070W WO 2004033729 A1 WO2004033729 A1 WO 2004033729A1
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WO
WIPO (PCT)
Prior art keywords
container
sample
environment
conducted
isolation
Prior art date
Application number
PCT/RU2003/000070
Other languages
French (fr)
Inventor
Andrei Zaslavskii
Original Assignee
Andrei Zaslavskii
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 Andrei Zaslavskii filed Critical Andrei Zaslavskii
Priority to EA200400341A priority Critical patent/EA005714B1/en
Priority to AU2003219628A priority patent/AU2003219628A1/en
Publication of WO2004033729A1 publication Critical patent/WO2004033729A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0278—Physical preservation processes
    • A01N1/0284—Temperature processes, i.e. using a designated change in temperature over time

Definitions

  • the present invention relates to biotechnology, in particular, to the methods of using genetic material of animal or vegetable nature, and can be used, for example, for the identification of humans.
  • Biomaterial any body tissue or liquid (muscle, piece of nail, hair, skin, bone, drop of blood, saliva or any other body liquid, vegetable tissue, vegetable juice etc.). This definition unifies all types of biological materials that can be further used for deriving information on the living body this material formerly belonged to.
  • Genetic code specially coded information on the sequence of amino acids composing the DNA of a biological object.
  • the genetic code and the genetic information coding system in nucleic acid molecules occur in biological objects in the form of a nucleotide sequence.
  • DNA deoxyribonucleic acid
  • DNA is a biological polymer consisting of numerous monomers called deoxyribonucleotides linked via the remainder of phosphoric acid in a sequence that is specific for each DNA.
  • the unique sequence of the deoxyribonucleotides in a DNA molecule is the genetic code.
  • identification can be made based on negligible pieces of human body by means of genetic analysis. This requires comparison sample, preferably piece of human body.
  • the object of the present invention is to provide a method of long term preservation of genetic samples with minimum costs.
  • the genetic sample preservation method comprising the steps of isolating the biological sample from the environment followed by low temperature exposure, preliminary sublimation of the sample placed in an open container, isolation of the inner surface of the container and the sample from the environment, the exposure being conducted at a temperature within +8 °C.
  • the sublimation is performed until the residual moisture content in the sample reduces to within 6 wt.% and the residual oxygen content in the sample reduces to within 4 wt.%.
  • the sample isolation from the environment is performed by soldering the container, filling the container with an inert gas, or combination of inert gas filling and soldering.
  • the samples can be stored in thermostats or exhausted salt mines. Usually, the samples are stored in glass, metallic, ceramic or polymer containers, but other durable containers can be envisioned.
  • thermostats or exhausted salt mines for the storage provides for unlimited biological sample storage time due to constant temperature and moisture.
  • a sample of biological material placed into a container is sublimated in standard sublimation equipment to remove microbiological contamination, moisture and oxygen.
  • the container is placed into a chemically neutral environment and covered. Vacuum is generated inside the container, or the air in the container is replaced for an inert gas or an inert liquid, liquid polymer, paraffin or another inert media.
  • the sample container is sealed using any known method (soldered, glued, welded, screwed or covered with a ground cover) to avoid the penetration of microorganisms, oxidizers and moisture into the container and their contact with the biological material sample.
  • the sample container is stored at a temperature close to 0 °C to avoid abrupt temperature drops. This provides better storage conditions because abrupt temperature drops impair the condition of biological materials.
  • Temperature can be stabilized by placing the container for long term storage into a cellar, a cave, a mine or deep into the water where the temperature is constant about 4 °C or into other places where this condition is satisfied.
  • the storage place should be protected from direct sunlight (ultraviolet radiation) and radiation, except the natural radioactive background.
  • Sets of biological material containers can be placed into larger containers to provide additional protection from the environment thus allowing safe storage and easier access to individual samples (containers).
  • Biological material containers can be marked or contain additional information.
  • a muscle tissue sample of a person working under high risk conditions is placed into an open glass container with a ground cover.
  • the open container is placed into a sublimation chamber. After the residual moisture and oxygen contents in the tissue reduce to 7 and 6 wt.% respectively, the container is removed from the sublimation chamber and filled to the cover level with silicon oil preliminarily heated to 120 °C and cooled down to room temperature.
  • silicon oil preliminarily heated to 120 °C and cooled down to room temperature.
  • the marked container is placed into a cave without temperature and humidity drops where the average temperature is in the range from +2 to +5 °C.
  • the storage life of a sample prepared as above in such a cave is more than 300 years (determined by extrapolation of temporal changes in the sample).
  • a nail bed sample of a test pilot was placed into a polyethylene container capable of being soldered.
  • the sample container is placed into a sublimation chamber to reduce the residual moisture and oxygen contents in the sample to 8 and 7 wt.% respectively, filled with dewatered carbon dioxide, soldered and marked with information on the nail bed donor.
  • the container and other similar containers are placed into a larger vinyl plastic container that is filler with argon and welded. Further the larger container is placed onto seabed at a depth of about 100 m where the water temperature is almost constantly equal to +4 °C. Extrapolation of experimental data suggests that the storage time of the sample is at least 280 years.
  • a sequoia wood sample was placed into a ceramic container, and the container was placed into a sublimation chamber. After the processing the residual moisture and oxygen contents in the sample reduced to 7 and 8 wt.% respectively, and no microorganisms remained on the container surface.
  • the inner volume of the container was filled with molten paraffin, and a ceramic cover was placed onto the hot paraffin to seal the contents of the container.
  • the container was further placed into an exhausted salt mine where the conditions were constant (+2 °C and 27% RH). Extrapolation of experimental data suggests that the storage time of the sample is more than 500 years.
  • the biological material preservation method allows infinitely long storage of samples for genetic analysis and cloning.

Abstract

The present invention relates to biotechnology, in particular, to the methods of using genetic material of animal or vegetable nature, and can be used, for example, for the identification of humans.

Description

Method of Preserving Biological Materials
The present invention relates to biotechnology, in particular, to the methods of using genetic material of animal or vegetable nature, and can be used, for example, for the identification of humans.
The following terms and definitions shall be used herein:
Biological material: any body tissue or liquid (muscle, piece of nail, hair, skin, bone, drop of blood, saliva or any other body liquid, vegetable tissue, vegetable juice etc.). This definition unifies all types of biological materials that can be further used for deriving information on the living body this material formerly belonged to.
Genetic code: specially coded information on the sequence of amino acids composing the DNA of a biological object. The genetic code and the genetic information coding system in nucleic acid molecules occur in biological objects in the form of a nucleotide sequence.
Genetic material: deoxyribonucleic acid (DNA).
DNA is a biological polymer consisting of numerous monomers called deoxyribonucleotides linked via the remainder of phosphoric acid in a sequence that is specific for each DNA. The unique sequence of the deoxyribonucleotides in a DNA molecule is the genetic code.
Today, pressing problem is identification of the remains of humans killed as a result of natural disasters, technogenic catastrophes, military actions etc.. Unfortunately, remains of biological objects that can be identified as human are often found in a condition that precludes their practical use for identification with conventional methods. The remains can be in the form of pieces of bodies without special signs sufficient for reliable person identification.
However, identification can be made based on negligible pieces of human body by means of genetic analysis. This requires comparison sample, preferably piece of human body.
Therefore for persons whose jobs involve significant risks it is preferable to have a bank of tissue samples that can be used for genetic analysis if remains of the body cannot be identified by any other means.
At present, there are several methods of preserving biological materials.
It has been suggested to preserve biological tissue samples by freezing (SU Inventor's Certificate 395439), embalming (SU Inventor's Certificate 1745765), paraffin sealing (RU Patent 2031114) and deposition of tissue cell monolayer on a carrier with further refrigerated storage (RU Patent 2014359).
All of the above methods fail to provide for long term storage of a biological material (during several years) because moisture and oxygen are present in the sample and microorganisms are present on the sample and surfaces. The presence of moisture and oxygen in the sample and the contact of the sample with oxygen eventually causes oxidation of the biological material and makes it inapplicable as reference for genetic identification. The closest counterpart of the method suggested here is the biological sample preservation methods by its isolation in an inert gas (SU Inventor's Certificate 1183537) for further storage.
Disadvantages of said method is the impossibility of long term storage due to the presence of moisture and oxygen in the sample.
The object of the present invention is to provide a method of long term preservation of genetic samples with minimum costs.
Said object can be achieved with the genetic sample preservation method comprising the steps of isolating the biological sample from the environment followed by low temperature exposure, preliminary sublimation of the sample placed in an open container, isolation of the inner surface of the container and the sample from the environment, the exposure being conducted at a temperature within +8 °C. Usually, the sublimation is performed until the residual moisture content in the sample reduces to within 6 wt.% and the residual oxygen content in the sample reduces to within 4 wt.%. Preferably, the sample isolation from the environment is performed by soldering the container, filling the container with an inert gas, or combination of inert gas filling and soldering. The samples can be stored in thermostats or exhausted salt mines. Usually, the samples are stored in glass, metallic, ceramic or polymer containers, but other durable containers can be envisioned.
Experiments have shown that during sample sublimation with moisture the sample loses oxygen, and all the microorganisms present on the container surface are eliminated. This lengthens the storage life of the sample. Further sample and container isolation from ambient moisture and oxygen also favors long term storage.
The use of thermostats or exhausted salt mines for the storage provides for unlimited biological sample storage time due to constant temperature and moisture.
The general embodiment of the present invention is as follows.
A sample of biological material placed into a container is sublimated in standard sublimation equipment to remove microbiological contamination, moisture and oxygen. The container is placed into a chemically neutral environment and covered. Vacuum is generated inside the container, or the air in the container is replaced for an inert gas or an inert liquid, liquid polymer, paraffin or another inert media. After that the sample container is sealed using any known method (soldered, glued, welded, screwed or covered with a ground cover) to avoid the penetration of microorganisms, oxidizers and moisture into the container and their contact with the biological material sample. The sample container is stored at a temperature close to 0 °C to avoid abrupt temperature drops. This provides better storage conditions because abrupt temperature drops impair the condition of biological materials. Temperature can be stabilized by placing the container for long term storage into a cellar, a cave, a mine or deep into the water where the temperature is constant about 4 °C or into other places where this condition is satisfied. The storage place should be protected from direct sunlight (ultraviolet radiation) and radiation, except the natural radioactive background. Sets of biological material containers can be placed into larger containers to provide additional protection from the environment thus allowing safe storage and easier access to individual samples (containers).
Biological material containers can be marked or contain additional information.
The present invention will be disclosed below using specific embodiments.
A muscle tissue sample of a person working under high risk conditions is placed into an open glass container with a ground cover. The open container is placed into a sublimation chamber. After the residual moisture and oxygen contents in the tissue reduce to 7 and 6 wt.% respectively, the container is removed from the sublimation chamber and filled to the cover level with silicon oil preliminarily heated to 120 °C and cooled down to room temperature. Experiments have shown that the sublimation conditions used in this method remove all the living microorganisms on the sample and container surfaces and remove moisture, oxygen and microorganisms in the silicon oil. The silicon oil filled container is covered with a ground cover and marked with information on the tissue donor. The marked container is placed into a cave without temperature and humidity drops where the average temperature is in the range from +2 to +5 °C. The storage life of a sample prepared as above in such a cave is more than 300 years (determined by extrapolation of temporal changes in the sample).
A nail bed sample of a test pilot was placed into a polyethylene container capable of being soldered. The sample container is placed into a sublimation chamber to reduce the residual moisture and oxygen contents in the sample to 8 and 7 wt.% respectively, filled with dewatered carbon dioxide, soldered and marked with information on the nail bed donor. The container and other similar containers are placed into a larger vinyl plastic container that is filler with argon and welded. Further the larger container is placed onto seabed at a depth of about 100 m where the water temperature is almost constantly equal to +4 °C. Extrapolation of experimental data suggests that the storage time of the sample is at least 280 years.
A sequoia wood sample was placed into a ceramic container, and the container was placed into a sublimation chamber. After the processing the residual moisture and oxygen contents in the sample reduced to 7 and 8 wt.% respectively, and no microorganisms remained on the container surface. The inner volume of the container was filled with molten paraffin, and a ceramic cover was placed onto the hot paraffin to seal the contents of the container. The container was further placed into an exhausted salt mine where the conditions were constant (+2 °C and 27% RH). Extrapolation of experimental data suggests that the storage time of the sample is more than 500 years.
The biological material preservation method allows infinitely long storage of samples for genetic analysis and cloning.

Claims

What is claimed is a
1. Genetic sample preservation method comprising the steps of isolating the biological sample from the environment followed by low temperature exposure, wherein said method further comprises preliminary sublimation of the sample placed in an open container, isolation of the inner surface of the container and the sample from the environment, the exposure being conducted at a temperature within +8 °C.
2. Method according to p. 1 wherein said sublimation is conducted until the residual moisture content reduces to within 6 wt.%.
3. Method according to p. 1 wherein said sublimation is conducted until the residual oxygen content reduces to within 4 wt.%.
4. Method according to any of pp. 1 through 3 wherein said isolation from the environment is conducted by soldering the container.
5. Method according to any of pp. 1 through 3 wherein said isolation from the environment is conducted by filling the container with an inert material.
6. Method according to any of pp. 1 through 3 wherein said isolation from the environment is conducted by filling the container with an inert material and further sealing.
7. Method according to any of pp. 1 through 6 wherein said container is stored in a thermostat.
8. Method according to any of pp. 1 through 6 wherein said container is stored in an exhausted salt mine.
9. Method according to any of pp. 1 through 8 wherein said container is in glass.
10. Method according to any of pp. 1 through 8 wherein said container is in polymer.
11. Method according to any of pp. 1 through 10 wherein said biological material container is placed into a larger container.
12.Method according to any of pp. 1 through 10 wherein said biological material container is marked and/or contains additional information.
PCT/RU2003/000070 2002-10-09 2003-02-28 Method of preserving biological materials WO2004033729A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EA200400341A EA005714B1 (en) 2002-10-09 2003-02-28 Method of preserving biological materials
AU2003219628A AU2003219628A1 (en) 2002-10-09 2003-02-28 Method of preserving biological materials

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
RU2002126937/15A RU2228617C2 (en) 2002-10-09 2002-10-09 Method for prolonged storage of biological material
RU2002126937 2002-10-09

Publications (1)

Publication Number Publication Date
WO2004033729A1 true WO2004033729A1 (en) 2004-04-22

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AU (1) AU2003219628A1 (en)
EA (1) EA005714B1 (en)
RU (1) RU2228617C2 (en)
WO (1) WO2004033729A1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2928517A1 (en) * 2008-03-11 2009-09-18 Imagene Sa INDUSTRIAL PROCESS FOR ENCAPSULATION OF BIOLOGICAL EQUIPMENT FOR AMBIENT TEMPERATURE CONSERVATION
CN103260399A (en) * 2010-08-24 2013-08-21 崔敏成 Business method by preserving genes and storing records during lifetime
EP3473094A1 (en) * 2017-10-18 2019-04-24 Centre National de la Recherche Scientifique CNRS High-fidelity long-term storage of biological material

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2703058C2 (en) * 2017-12-08 2019-10-15 Владимир Артурович Хачумов Method of storing dna-containing plant material in a wide temperature range

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5856102A (en) * 1997-02-26 1999-01-05 Bierke-Nelson; Diane Lynn Home/self-storage to improve DNA banking
WO1999057264A1 (en) * 1998-05-06 1999-11-11 Sophie Tuffet Method for prolonged storage of dna molecules and packaging implementing said method

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5856102A (en) * 1997-02-26 1999-01-05 Bierke-Nelson; Diane Lynn Home/self-storage to improve DNA banking
WO1999057264A1 (en) * 1998-05-06 1999-11-11 Sophie Tuffet Method for prolonged storage of dna molecules and packaging implementing said method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BURGASOVA P. N., 1978, MOSCOW, MEDITSINA, article "Rukovodstvo po vaktsionnomu I syvorotochnomu delu" *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2928517A1 (en) * 2008-03-11 2009-09-18 Imagene Sa INDUSTRIAL PROCESS FOR ENCAPSULATION OF BIOLOGICAL EQUIPMENT FOR AMBIENT TEMPERATURE CONSERVATION
WO2009115761A2 (en) * 2008-03-11 2009-09-24 Imagene Industrial method for encapsulation of biological material with a view to storage at ambient temperature
WO2009115761A3 (en) * 2008-03-11 2009-12-10 Imagene Industrial method for the encapsulation of a biological material with a view to storage at ambient temperature, including an encapsulation vacuum sealing test
JP2011512860A (en) * 2008-03-11 2011-04-28 イマジェネ Industrial method of encapsulation intended for storage of biological materials at room temperature, including hermeticity testing of encapsulation under vacuum
CN103260399A (en) * 2010-08-24 2013-08-21 崔敏成 Business method by preserving genes and storing records during lifetime
EP3473094A1 (en) * 2017-10-18 2019-04-24 Centre National de la Recherche Scientifique CNRS High-fidelity long-term storage of biological material
WO2019077048A1 (en) * 2017-10-18 2019-04-25 Centre National De La Recherche Scientifique (Cnrs) High-fidelity long-term storage of biological material

Also Published As

Publication number Publication date
AU2003219628A1 (en) 2004-05-04
EA200400341A1 (en) 2004-08-26
RU2228617C2 (en) 2004-05-20
EA005714B1 (en) 2005-04-28

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