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WO2010008059A1 - Procédé de stockage de cellules et dispositif pour sa mise en œuvre - Google Patents

Procédé de stockage de cellules et dispositif pour sa mise en œuvre Download PDF

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Publication number
WO2010008059A1
WO2010008059A1 PCT/JP2009/062918 JP2009062918W WO2010008059A1 WO 2010008059 A1 WO2010008059 A1 WO 2010008059A1 JP 2009062918 W JP2009062918 W JP 2009062918W WO 2010008059 A1 WO2010008059 A1 WO 2010008059A1
Authority
WO
WIPO (PCT)
Prior art keywords
storage
cells
pressure
cell
container
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2009/062918
Other languages
English (en)
Japanese (ja)
Inventor
昭夫 清水
進一 早乙女
一行 中嶋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
TAMA-TLO Ltd
Tama TLO Co Ltd
Soka University
Original Assignee
TAMA-TLO Ltd
Tama TLO Co Ltd
Soka University
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 TAMA-TLO Ltd, Tama TLO Co Ltd, Soka University filed Critical TAMA-TLO Ltd
Publication of WO2010008059A1 publication Critical patent/WO2010008059A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • A01N1/10—Preservation of living parts
    • A01N1/14—Mechanical aspects of preservation; Apparatus or containers therefor
    • A01N1/146—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving
    • 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
    • A01N1/10—Preservation of living parts
    • A01N1/16—Physical preservation processes
    • A01N1/165—Pressure processes, e.g. following predefined pressure changes over time

Definitions

  • Cells collected from living organisms can be cultured in vitro to examine their physiological activities and functions, used as samples for pharmaceutical development, and used for treatments such as transplantation. Yes. Particularly in recent years, due to social criticism of animal experiments, it has become an international trend to minimize animal experiments and replace them with cultured cells.
  • Patent Document 1 a method for storing food and microorganisms under high pressure of 500 to 10000 atm (about 50 to 1000 MPa) in order to store them without freezing has been disclosed.
  • Patent Document 2 a storage method is disclosed under a low temperature and a high pressure of 70 to 1000 atmospheres (about 7 to 100 MPa) in combination with a storage solution.
  • the storage temperature is higher than the crystallization temperature of intracellular water to be stored, the cell activity optimum temperature or less, and the storage pressure is 0.11 to This is a storage method of 10 MPa.
  • the optimum temperature of the cell activity means a temperature suitable for the life activity of the cell, and the temperature varies depending on the cell type. For example, in the case of an animal cell, the body temperature of the animal is That is true.
  • the storage temperature is 1 ° C. to 37 ° C.
  • the storage pressure is 0.11 to 6.5 MPa.
  • the cells are seeded or suspended in a medium that is liquid under the aforementioned storage conditions.
  • the pressure medium is a liquid.
  • the cell storage device includes a pressurized container capable of maintaining pressure by filling with a pressure medium, and can be accommodated in the pressurized container.
  • a pressurized container capable of maintaining pressure by filling with a pressure medium, and can be accommodated in the pressurized container.
  • One or two or more cell storage containers capable of pressurizing cells enclosed inside by having a material that can be transmitted to a substance.
  • the cell storage device adjusts the pressure in the pressurized container by sending the pressure medium into the pressurized container by a fluid transfer device.
  • the storage container is placed in a container for applying pressure (hereinafter referred to as “pressurized container”), and a gas or liquid is used as a pressure medium in the storage container. Apply pressure to the cells.
  • the number of storage containers stored in the pressurized container may be one, or two or more may be used when several types of cells are desired to be stored at the same time.
  • the number of storage containers stored in the pressurized container is preferably 100 or less from the design of the apparatus.
  • the cells placed in the storage container as described above are stored at a temperature higher than the crystallization temperature of the water in the cells, not more than the optimum temperature for cell activity, and a pressure of 0.11 to 10 MPa.
  • the water crystallization temperature varies depending on the cell type, but is generally 0 to -10 ° C. If the temperature is lower than the crystallization temperature, the water in the cell may crystallize and destroy the cell. If the temperature is higher than the optimum temperature for the activity, it is generally an unsuitable environment for the cell and the cell viability decreases. is there.
  • the storage pressure is preferably 0.11 MPa or more in terms of cell viability, and more preferably 0.15 MPa or more in consideration of the risk of pressure leakage during storage.
  • the maximum pressure is preferably 6.5 MPa or less, more preferably 5.5 MPa or less, from the viewpoint of maintaining the cell viability for a long period of time.
  • the size of the cell storage device of the present invention is not particularly limited as long as it is theoretically possible in terms of production, but for the purpose of cell storage, a cell storage container capacity of 0.01 mL to several L is practical. Preferably 0.1 mL to 1 L.
  • the pressurized container is designed in accordance with the size of the storage container and the number of the storage containers stored in the pressurized container.
  • the liquid feed pump 2 and the pressurized container 4 are connected by a Teflon (registered trademark) tube through a valve 5, and after storing the storage container 3 in the pressurized container 4 and closing the open / close hatch, the valve 6 is turned on.
  • the valve 6 In an open state, the water in the storage tank 1 is fed into the pressurized container 4 by the liquid feed pump 2. Alternatively, water may be fed into the pressurized container before storing the storage container.
  • the valve 6 is closed. In this state, while monitoring the pressure in the pressurized container 4 with the pressure indicator 8, further water is fed in until the desired pressure is reached. At this time, it is desirable that water as the pressure medium is maintained at a desired storage temperature.
  • the storage container 3 and the pressurized container 4 are both cylindrical.
  • the storage container 3 may have a spherical shape or a bag shape such as an infusion bag.
  • a rectangular parallelepiped or a cube having a hollow inside, a so-called cooler box-like shape may be used. That is, for example, a storage method may be used in which a plurality of drip bag-like storage containers are arranged in a cooler box-like pressurized container.
  • the valve 5 or 6 or both may be opened to discharge the liquid pressure medium (water in this embodiment) to return the pressure to atmospheric pressure.
  • the liquid pressure medium may be discharged at a desired speed using a liquid feed pump. If the temperature at the time of cell storage and the temperature at the time of treatment after storage are different, the temperature may be adjusted before returning the pressure to atmospheric pressure, or the temperature may be adjusted after returning the pressure to atmospheric pressure. Although it is good, it is preferable to return to atmospheric pressure after adjusting the temperature.
  • Comparative Examples 1 and 2 The sample was stored under the same conditions as in Example 1 except that the storage pressure was 0.1 or 14 MPa (Comparative Examples 1, 2, and so on). As a result, the cell survival rate was about 20% under 0.1 MPa and less than 10% at 14 MPa.
  • Comparative Examples 5-6 It was stored under the same conditions as in Example 6 except that the storage pressure was 0.1 or 11 MPa. As a result, the cell viability in each case was less than 10%.
  • Examples 8-11 It was stored under the same conditions as in Example 6 except that the storage temperature was 4 ° C. and the storage pressure was 0.2, 0.5, 1.0, or 6.5 MPa. As a result, the cell viability when the storage pressure was 0.2, 0.5, or 1.0 MPa was good at 60% or more. In the case of 6.5 MPa, it was about 50%.
  • Comparative Example 7 It was stored under the same conditions as in Example 6 except that the storage temperature was 4 ° C. and the storage pressure was 0.1 MPa. As a result, the cell survival rate was almost 0%.
  • Comparative Examples 9-10 It was stored under the same conditions as in Example 14 except that the storage pressure was 0.1 or 11 MPa. As a result, the cell viability in each case was almost 0%.
  • Table 2 shows the results for a storage period of 1 day.
  • Table 3 shows the results for a storage period of 4 days.
  • Table 4 shows the results for a storage period of 5 days.
  • Cells are subjected to experiments for various purposes in a wide range such as medical fields, pharmaceutical fields, livestock fields, and academic fields such as biochemistry. It also plays an important role in transplantation treatment.
  • the experiment may be temporarily interrupted to be stored. In such cases, simply refrigeration not only lowers cell viability, but some cells cannot be preserved.
  • the cells are frozen or a storage solution is used, as described above, there are cases where the method cannot be employed due to lack of rapid storage and resumption of experiments.
  • the storage method of the present invention these disadvantages can be solved, and storage and experiments can be resumed quickly and easily. For example, primary cultured cells that maintain their properties in vivo are desired to be used for experiments quickly. It is possible to meet the demand for cell preservation in a wide range of fields.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un procédé de stockage de cellules par lequel les exigences pour la rapidité et la commodité de traitement des cellules avant et après le stockage et pour la facilité de transport de celles-ci peuvent être satisfaites tout en maintenant le taux de survie des cellules à un niveau élevé, et un dispositif pour la mise en œuvre de ce procédé. L'invention porte sur un procédé de stockage de cellules dans lequel la température de stockage est régulée de façon à être supérieure à la température de cristallisation de l'eau dans les cellules devant être stockées et non supérieure à la température d'action optimale des cellules alors que la pression de stockage est régulée entre 0,11 et 10 MPa. L'invention porte également sur un dispositif permettant le stockage de cellules, comprenant un récipient mis sous pression qui peut être maintenu sous une pression définie lorsqu'il est conditionné avec un milieu de pression, et un ou plusieurs récipients de stockage de cellules qui peuvent être placés dans le récipient mis sous pression et ont une aptitude à transférer la pression de l'extérieur à une matière contenue dans ceux-ci de telle sorte que des cellules enfermées dans ceux-ci peuvent être mises sous pression.
PCT/JP2009/062918 2008-07-18 2009-07-16 Procédé de stockage de cellules et dispositif pour sa mise en œuvre Ceased WO2010008059A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2008-187304 2008-07-18
JP2008187304A JP2010022269A (ja) 2008-07-18 2008-07-18 細胞の保存方法及びその装置

Publications (1)

Publication Number Publication Date
WO2010008059A1 true WO2010008059A1 (fr) 2010-01-21

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PCT/JP2009/062918 Ceased WO2010008059A1 (fr) 2008-07-18 2009-07-16 Procédé de stockage de cellules et dispositif pour sa mise en œuvre

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JP (1) JP2010022269A (fr)
WO (1) WO2010008059A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2012192061A (ja) * 2011-03-17 2012-10-11 Mie Univ 前十字靭帯組織およびその製造方法
CN103327839A (zh) * 2011-01-20 2013-09-25 弥通雅贸易有限公司 低温保存方法及低温保存容器

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2015050992A (ja) * 2013-08-07 2015-03-19 ｉＰＳアカデミアジャパン株式会社 多能性幹細胞の輸送方法
US10669525B2 (en) 2014-09-29 2020-06-02 Public University Corporation Yokohama City University Method for producing three-dimensional cell aggregates

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4712445U (fr) * 1971-03-11 1972-10-14
WO2000025580A1 (fr) * 1998-10-30 2000-05-11 Hyperbaric Systems Procede et appareil permettant de conserver des matieres biologiques
WO2000053008A1 (fr) * 1999-03-11 2000-09-14 Hyperbaric Systems Compositions et procedes de conservation de plaquettes
WO2007060608A2 (fr) * 2005-11-22 2007-05-31 Sempre Ltd. Augmentation de la viabilite et de la tolerance au stress d'un materiau biologique viable

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4712445U (fr) * 1971-03-11 1972-10-14
WO2000025580A1 (fr) * 1998-10-30 2000-05-11 Hyperbaric Systems Procede et appareil permettant de conserver des matieres biologiques
WO2000053008A1 (fr) * 1999-03-11 2000-09-14 Hyperbaric Systems Compositions et procedes de conservation de plaquettes
WO2007060608A2 (fr) * 2005-11-22 2007-05-31 Sempre Ltd. Augmentation de la viabilite et de la tolerance au stress d'un materiau biologique viable

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
AKIO SHIMIZU ET AL.: "Dobutsu Plankton Oyobi Tamago no Fuka ni Taisuru Atsuryoku no Eikyo Sokuteiyo Koatsu System", THE REVIEW OF HIGH PRESSURE SCIENCE AND TECHNOLOGY, vol. 16, no. 4, 2006, pages 381 - 383, Retrieved from the Internet <URL:http://www.jstage.jst.go.jp/article/jshpreview/16/4/381/_pdf/-char/ja> *
AKIO SHIMIZU ET AL.: "Kaatsu ni yoru Saibo no Kan'i Hozon Hoho -Mochihakobi Kano na Yuso Yoki", KOIKI TAMA(TAMA) CHIIKI NO DAIGAKU HATSU, SHIN GIJUTSU SETSUMEIKAI, TOJITSU HAIFU SHIRYO, 29 August 2008 (2008-08-29), Retrieved from the Internet <URL:http://jstshingi.jp/abst/p/08/818/tamal.pdf> *
AKIO SHIMIZU ET AL.: "Koatsu Shori ni yoru Kosui no Jukusei Sokushin", MIRAI O HIRAKU KOATSURYOKU KAGAKU GIJUTSU SEMINAR SERIES (33), 6 March 2008 (2008-03-06), pages 20 - 25 *
IKUO SERA ET AL.: "Biseibutsu no Atsuryoku to Ondo ni Taisuru Taisei Jikken", BIOLOGICAL SCIENCES IN SPACE, vol. 12, no. 3, 1998, pages 232 - 233 *
IKUSHI IWATANI ET AL.: "Seisui Kaatsu ni yoru Kakushu Dobutsu Saibo no Zoshoku Seigyo", CSJ: THE CHEMICAL SOCIETY OF JAPAN DAI 89 SHUNKI NENKAI (2009) KOEN YOKOSHU II, 13 March 2009 (2009-03-13), pages 1535 *
MASAHIRO TANIUCHI ET AL.: "Koatsuryoku Shori ni yoru Kosui no Jukusei Kikan Tansyuku ni Kansuru Kenkyu", NEW FOOD IND., vol. 49, no. 12, 2007, pages 1 - 5 *
TETSUYA MIWA: "Baiyo Saibo no Koatsuryoku ni Taisuru Oto Tokusei", CSJ: THE CHEMICAL SOCIETY OF JAPAN DAI 81 SHUNKI NENKAI 2002 NEN KOEN YOKOSHU II, 2002, pages 872 *
TETSUYA MIWA: "Koatsuka ni Okeru Baiyo Saibo no Sonoba Kansatsu", JAPAN SOCIETY FOR BIOSCIENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY 2001 NENDO TAIKAI KOEN YOSHISHU, 2001, pages 535 *
TOMOKO YOSHIKI ET AL.: "Kaisan Dobutsu Plankton Kenkyu no Tameno Atsuryoku Sochi no Kaihatsu", 2005 NENDO THE OCEANOGRAPHIC SOCIETY OF JAPAN SHUKI TAIKAI KOEN YOSHISHU, 2005, pages 197, 341 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103327839A (zh) * 2011-01-20 2013-09-25 弥通雅贸易有限公司 低温保存方法及低温保存容器
JP2012192061A (ja) * 2011-03-17 2012-10-11 Mie Univ 前十字靭帯組織およびその製造方法

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