JP2009118820A - Compact rotary culture system and culture vessel - Google Patents

Compact rotary culture system and culture vessel Download PDF

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JP2009118820A
JP2009118820A JP2007298869A JP2007298869A JP2009118820A JP 2009118820 A JP2009118820 A JP 2009118820A JP 2007298869 A JP2007298869 A JP 2007298869A JP 2007298869 A JP2007298869 A JP 2007298869A JP 2009118820 A JP2009118820 A JP 2009118820A
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test tube
culture
rotary
present
rotary culture
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Tomoyuki Kawase
知之 川瀬
Kunio Hane
邦夫 羽根
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DEUCE KK
Niigata University NUC
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DEUCE KK
Niigata University NUC
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/08—Flask, bottle or test tube
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/24—Gas permeable parts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/38—Caps; Covers; Plugs; Pouring means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/10—Rotating vessel
    • C12M27/12—Roller bottles; Roller tubes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/20—Material Coatings
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/46—Means for fastening

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inexpensive, space-saving and readily operable culture system for three-dimensional high-density cell culture so that even medium and small hospitals and clinics are capable of bearing a burden in order to further spread regenerative therapy. <P>SOLUTION: A rotary culture system includes a test tube 10 with a cap 11 characterized by having one or two or more through-holes 13 in the sidewall of the test tube in which the through-holes are closed with an air-permeable and water-impermeable membrane 14, a rotary culture apparatus capable of installing the one or two test tubes thereon, an airtight vessel for housing the test tubes and the rotary culture apparatus, a controller for gas composition and/or pressure in the airtight vessel, and a controller for temperature in the airtight vessel. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

本発明は、コンパクト回転培養システム及び培養容器に関し、具体的には、再生医療に用いるヒト正常細胞を３次元高密度培養法で増殖させるためのコンパクトな回転培養システム及び培養容器に関する。   The present invention relates to a compact rotating culture system and a culture container, and more particularly to a compact rotating culture system and a culture container for growing normal human cells used for regenerative medicine by a three-dimensional high-density culture method.

再生医療においてヒト正常細胞又は組織を移植するためには、３次元高密度細胞培養を行う必要がある。例えば、特許文献１には、多孔質ハイドロキシアパタイト・ブロック体に細胞を播種、培養し石灰化を誘導することを特徴とする培養人工骨の作成方法が説明される。
特願２００７−０５１１５３号明細書 In order to transplant human normal cells or tissues in regenerative medicine, it is necessary to perform three-dimensional high-density cell culture. For example, Patent Document 1 describes a method for producing a cultured artificial bone, characterized in that cells are seeded and cultured in a porous hydroxyapatite block body to induce calcification.
Japanese Patent Application No. 2007-051153

しかし、従来の３次元高密度細胞培養用培養システムは汎用型バイオリアクタを使用するため、装置設備の設置に場所をとり、大量の培養液を消費し、操作及びメンテナンスが煩雑であり、装置及び消耗品の経費が高額になるという欠点がある。かかるバイオリアクタを使用せずに、通常のＣＯ２インキュベータ内に攪拌器と培養用フラスコを設置したり、回転培養装置を設置することも考えられる。その場合でも、攪拌器や回転培養装置を収容可能なサイズのＣＯ２インキュベータはＣＯ２ボンベを含めて広い占有面積（最低でも３ｍ２）を必要とし、導入コストも１００万円程度は必要である。これは施設及び予算の制約が厳しい中小病院や診療所にとって再生医療を導入するうえで大きな障害である。 However, since the conventional culture system for three-dimensional high-density cell culture uses a general-purpose bioreactor, it takes a lot of space to install the equipment, consumes a large amount of culture solution, and is complicated in operation and maintenance. There is a disadvantage that the cost of consumables becomes high. It is also conceivable to install a stirrer and a culture flask in a normal CO 2 incubator without using such a bioreactor, or install a rotary culture apparatus. Even in such a case, a CO 2 incubator of a size that can accommodate a stirrer and a rotary culture apparatus requires a large occupied area (at least 3 m 2 ) including the CO 2 cylinder, and an introduction cost of about 1 million yen is necessary. . This is a major obstacle to the introduction of regenerative medicine for small and medium-sized hospitals and clinics where facilities and budget constraints are severe.

そこで、再生医療のさらなる普及を図るため、中小病院や診療所でも負担できるように、低コストで、省スペースかつ操作が容易な３次元高密度細胞培養用培養システムを開発する必要がある。   Therefore, in order to further promote regenerative medicine, it is necessary to develop a low-cost, space-saving and easy-to-operate culture system for three-dimensional high-density cell culture that can be borne by small and medium hospitals and clinics.

本発明は、キャップ付の試験管であって、該試験管の側壁には１個又は２個以上の貫通孔が設けられ、該貫通孔は、通気性があり、かつ、透水性のない膜で閉塞されていることを特徴とする試験管を提供する。   The present invention is a test tube with a cap, wherein one or two or more through holes are provided in a side wall of the test tube, and the through hole is a membrane having air permeability and no water permeability. A test tube characterized in that it is closed with is provided.

本発明は、本発明の試験管を１本又は２本以上装架することができるローテータと、該ローテータを駆動するモータとを含むことを特徴とする回転培養装置を提供する。   The present invention provides a rotating culture apparatus comprising a rotator capable of mounting one or more test tubes of the present invention, and a motor for driving the rotator.

本発明の回転培養装置は電源としての電池を含む場合がある。   The rotary culture device of the present invention may include a battery as a power source.

本発明は、前記本発明の試験管と、前記本発明の回転培養装置と、前記試験管及び回転培養装置を収容する気密容器と、該気密容器内のガス組成及び／又は圧力の制御装置と、前記気密容器内温度の制御装置とを含む回転培養システムを提供する。   The present invention includes a test tube according to the present invention, a rotary culture device according to the present invention, an airtight container containing the test tube and the rotary culture device, and a gas composition and / or pressure control device in the airtight container; And a rotary culture system including the air-tight container internal temperature control device.

本発明の回転培養システムにおいて、前記試験管及び気密容器はプラスチックでできており、前記ガス組成及び圧力の制御装置はガス発生剤である場合がある。   In the rotary culture system of the present invention, the test tube and the airtight container may be made of plastic, and the gas composition and pressure control device may be a gas generating agent.

以下に本発明の実施例によって、本発明を詳細に説明するが、本発明はこれらの実施例により何ら制限されるものではない。   EXAMPLES The present invention will be described in detail below with reference to examples of the present invention, but the present invention is not limited to these examples.

図１は本発明の試験管１０の透視図である。試験管１０は、キャップ１１と、試験管本体１２とを含む。キャップ１１は試験管本体１２に固定されて試験管内部を無菌状態に保つ。キャップ１１はねじ込み式で試験管本体１２に固定される場合がある。キャップ１１は、可撓性又は弾性のある材料でできていて、試験管本体１２との摩擦係合によって試験管本体１２に固定されるプッシュオン式の場合もある。   FIG. 1 is a perspective view of a test tube 10 of the present invention. The test tube 10 includes a cap 11 and a test tube body 12. The cap 11 is fixed to the test tube body 12 to keep the inside of the test tube in a sterile state. The cap 11 may be fixed to the test tube body 12 by screwing. The cap 11 is made of a flexible or elastic material, and may be a push-on type that is fixed to the test tube body 12 by frictional engagement with the test tube body 12.

本発明の試験管１０は、細胞培養に使用できるものであればいかなる材料でできていてもかまわないが、プラスチックでできていることが好ましい。好ましい試験管は、当業者に周知の使い捨て可能なプラスチック製品と同様に製造される。試験管１０のサイズはいかなるものでもかまわないが、汎用の１５ｍＬ又は５０ｍＬの使い捨てプラスチック製試験管と互換性のある外部寸法を有することが好ましい。試験管本体１２の内壁は、培養される細胞が接着できるように表面処理を施される場合がある。あるいは試験管本体１２の内壁は、前記細胞が接着できないような材質でできているか、細胞接着を阻害するように表面処理を施される場合がある。ヒト正常上皮又は繊維芽細胞のように基質との接着が培養下での増殖に必要であるか、あるいは、望ましい細胞を培養する際には、基質面積を確保するために、細胞が接着できるように処理が施された固体支持体を培養液に懸濁することが好ましい。   The test tube 10 of the present invention may be made of any material as long as it can be used for cell culture, but is preferably made of plastic. Preferred test tubes are manufactured similar to disposable plastic products well known to those skilled in the art. The test tube 10 can be of any size, but preferably has external dimensions compatible with a general purpose 15 mL or 50 mL disposable plastic test tube. The inner wall of the test tube body 12 may be subjected to surface treatment so that cultured cells can adhere. Alternatively, the inner wall of the test tube body 12 may be made of a material that prevents the cells from adhering, or may be subjected to a surface treatment so as to inhibit cell adhesion. Adhesion with the substrate is necessary for growth in culture, such as human normal epithelium or fibroblasts, or when culturing desired cells, the cells can adhere to ensure substrate area It is preferable to suspend the solid support subjected to the treatment in the culture solution.

本発明の試験管１０の本体１２には、試験管内外のガス交換のために１個又は２個以上の貫通孔１４が設けられる。該貫通孔が設けられる位置は、試験管本体１２の側壁である。貫通孔１３は膜１４で閉塞される。貫通孔１３の孔径、数及び配置は、膜１４によって試験管内部へのガス交換はできるが、試験管内部の培養液の漏出が起こらず、かつ、試験管内部の無菌状態が維持されることを条件としていかなるものであってもかまわない。   The main body 12 of the test tube 10 of the present invention is provided with one or two or more through holes 14 for gas exchange inside and outside the test tube. The position where the through hole is provided is the side wall of the test tube body 12. The through hole 13 is blocked by the film 14. The diameter, number and arrangement of the through-holes 13 are such that gas can be exchanged into the test tube by the membrane 14, but the culture solution inside the test tube does not leak and the aseptic condition inside the test tube is maintained. Any condition is acceptable.

本発明の試験管本体１２の貫通孔１３を閉塞する膜１４は、通気性があり、かつ、透水性のないことを条件として、いかなるものであってもかまわない。好ましい膜は、米国マサチューセッツ州ボストン市のダイバーシファイド・バイオテク（Diversified Biotech）社のカタログ番号ＢＥＭ−１のように、通気性があって、透水性がないだけでなく、膜の一方の面が無菌で、かつ、粘着性があって、貫通孔１３を覆うように封止できるものである。しかしながら、膜１４は試験管本体の外壁又は内壁に圧着又は融着される場合がある。   The membrane 14 that closes the through hole 13 of the test tube main body 12 of the present invention may be anything as long as it is air permeable and not water permeable. A preferred membrane is not only breathable and non-permeable, such as Diversified Biotech catalog number BEM-1 in Boston, Massachusetts, USA. It is aseptic and sticky, and can be sealed so as to cover the through hole 13. However, the membrane 14 may be crimped or fused to the outer or inner wall of the test tube body.

図２は本発明の回転培養装置２０の透視図である。本発明の回転培養装置２０は、本発明の試験管１０を１本又は２本以上、好ましくは、２〜６本装架して回転することにより、試験管１０内部の培養液を攪拌させてガス交換を促進させる。回転培養装置２０は、ローテータ２１と、電動モータ２２からローテータ２１への駆動伝達機構とを含む。該駆動伝達機構は、プーリーとワイヤ又はベルトとを用いる方式の他、歯車とチェーンとを用いる方式や、歯車の組合せによる方式など、当業者に周知のいかなる駆動伝達機構を用いてもかまわない。ローテータ２１の回転速度は特に限定されないが、４〜８ｒｐｍの速度で回転することが好ましい。ローテータ２１は図２に示すような単一の棒又は筒状の形状をとる場合の他、中空の筒か、回転軸に平行に配置された複数の棒かの形状をとる場合（図示されない）がある。ローテータが単一の棒又は筒状の形状をとる場合には、試験管１０をローテータ２１に装架する手段は、クランプ等で試験管の側面を把持する態様や、ローテータ２１の回転軸に対して放射方向に延びる部材が、試験管のキャップと試験管の底部とを保持する態様（図示されない）をとる。これらの態様では、電動モータ２２への負担を低減するために、2本又は３本以上の試験管を回転軸に対して対称に配置することが好ましい。本発明の試験管かこれと同じ重量の物体をバランス用に装架する場合もある。ローテータが、中空の筒か、回転軸に平行に配置された複数の棒かの形状をとる場合には、１本の試験管を、該試験管の長手方向の軸が回転軸に一致するように装架することができる。また、本発明の回転培養装置のローテータは、本発明の試験管に外接して該試験管を摺動させて回転させる態様をとることもある。試験管の回転培養装置２０は、さらに、電源としての電池２３を含む場合がある。電池２３を含む回転培養装置２０は密閉容器に収容できるため好ましい。電池２３は充電式の場合がある。電池２３は、回転培養装置を少なくとも１日間、好ましくは３日間連続して運転することができることが望ましい。また、回転培養装置２０は、培養液又は電池の交換のための一時的停止を挟んで、少なくとも１ヶ月、好ましくは2ヶ月ないし３ヶ月の間連続して運転することができる程度の信頼性及び耐久性があることが好ましい。   FIG. 2 is a perspective view of the rotary culture apparatus 20 of the present invention. The rotary culture apparatus 20 of the present invention stirs the culture solution inside the test tube 10 by mounting one or two or more test tubes 10 of the present invention, preferably 2 to 6 and rotating them. Promote gas exchange. The rotary culture device 20 includes a rotator 21 and a drive transmission mechanism from the electric motor 22 to the rotator 21. The drive transmission mechanism may be any drive transmission mechanism known to those skilled in the art, such as a system using a pulley and a wire or a belt, a system using a gear and a chain, and a system using a combination of gears. The rotation speed of the rotator 21 is not particularly limited, but it is preferable to rotate at a speed of 4 to 8 rpm. The rotator 21 has a single rod or cylindrical shape as shown in FIG. 2, or a hollow tube or a plurality of rods arranged in parallel to the rotation axis (not shown). There is. When the rotator has a single rod or cylindrical shape, the means for mounting the test tube 10 on the rotator 21 is such that the side surface of the test tube is gripped by a clamp or the like, or the rotation axis of the rotator 21 The radially extending member takes a form (not shown) for holding the cap of the test tube and the bottom of the test tube. In these aspects, in order to reduce the burden on the electric motor 22, it is preferable to arrange two or three or more test tubes symmetrically with respect to the rotation axis. In some cases, the test tube of the present invention or an object of the same weight is mounted for balancing. When the rotator is in the form of a hollow cylinder or a plurality of bars arranged in parallel to the rotation axis, one test tube is arranged so that the longitudinal axis of the test tube coincides with the rotation axis. It can be mounted on. Further, the rotator of the rotating culture apparatus of the present invention may take a form in which the test tube is circumscribed and rotated by sliding the test tube. The test tube rotating culture apparatus 20 may further include a battery 23 as a power source. The rotary culture apparatus 20 including the battery 23 is preferable because it can be accommodated in a sealed container. The battery 23 may be rechargeable. It is desirable that the battery 23 can operate the rotary culture apparatus continuously for at least 1 day, preferably 3 days. In addition, the rotary culture apparatus 20 has such a reliability that it can be continuously operated for at least one month, preferably two to three months, with a temporary stop for replacement of the culture medium or battery. Durability is preferred.

図３は本発明の回転培養システム６０の断面模式図である。本発明の回転培養システム６０は、１本又は２本以上の試験管１０を装架した回転培養装置２０が収容された気密容器３０と、気密容器３０内のガス組成及び／又は圧力の制御装置４０と、気密装置３０内の温度の制御装置５０とを含む。気密容器３０内のガス組成は、水蒸気、酸素、窒素及び／又は二酸化炭素の濃度を任意に調整し維持できるものであればいかなるものでもかまわない。ガス組成及び／又は圧力の制御装置４０は、例えば、気密容器３０の容積に対して所定のガス濃度になる量のガス発生剤の場合がある。あるいは、特定のガス組成、例えば水蒸気又は酸素を吸収するガス吸収剤の場合がある。ガス組成及び／又は圧力の制御装置４０は、図３に示すように気密容器３０の内部に収容される場合の他、気密容器３０の外部にあって、気密容器３０と連絡する圧力及び／又はガス流量の調節器と、気密容器３０内部の気圧を調整するための通気口又は圧力弁と、酸素、窒素及び／又は二酸化炭素の高圧ガスボンベとを含む場合（図示されない）がある。気密容器３０内の温度の制御装置５０は、気密容器３０全体を収容するインキュベータの場合がある。本発明のガス組成及び／又は圧力の制御装置は、本発明の温度の制御装置と一体となったＣＯ２インキュベータとして、１個又は２個以上の気密容器３０を収容する場合がある。この場合、気密容器３０はガス交換のために貫通孔が設けられることがある。前記気密容器の貫通孔は、通気性のある膜で閉塞されることもある。 FIG. 3 is a schematic cross-sectional view of the rotary culture system 60 of the present invention. The rotary culture system 60 of the present invention includes an airtight container 30 in which a rotary culture apparatus 20 in which one or more test tubes 10 are mounted, and a gas composition and / or pressure control device in the airtight container 30. 40 and a temperature control device 50 in the hermetic device 30. The gas composition in the hermetic container 30 may be anything as long as the concentration of water vapor, oxygen, nitrogen and / or carbon dioxide can be arbitrarily adjusted and maintained. The gas composition and / or pressure control device 40 may be, for example, a gas generating agent in an amount that provides a predetermined gas concentration with respect to the volume of the hermetic container 30. Alternatively, it may be a gas absorbent that absorbs a specific gas composition, such as water vapor or oxygen. As shown in FIG. 3, the gas composition and / or pressure control device 40 is located outside the hermetic container 30 as shown in FIG. 3 and is located outside the hermetic container 30 and communicates with the hermetic container 30. There may be cases (not shown) including a gas flow rate regulator, a vent or pressure valve for adjusting the pressure inside the hermetic vessel 30, and a high pressure gas cylinder of oxygen, nitrogen and / or carbon dioxide. The temperature control device 50 in the hermetic container 30 may be an incubator that accommodates the entire hermetic container 30. The gas composition and / or pressure control device of the present invention may contain one or more hermetic containers 30 as a CO 2 incubator integrated with the temperature control device of the present invention. In this case, the airtight container 30 may be provided with a through hole for gas exchange. The through hole of the hermetic container may be blocked with a breathable membrane.

本発明の回転培養装置２０のローテータ２１の回転軸は水平方向と垂直方向との間のいずれの方向にあってもよい。試験管１０の長手方向の軸とローテータ２１の軸とは、平行の場合と、交差する場合と、同一平面には属さない場合とがある。好ましくは、本発明の試験管１０は、図２に示すように、水平な回転軸を有するローテータを含む回転培養装置に装架される。この場合には、ローテータの回転につれて試験管１０の内部の培養液は試験管本体の側壁全周に接するように移動する。本発明の試験管１０の長手方向の軸がローテータの回転軸と交差する場合又は同一平面に属さない場合には、ローテータの回転に伴って試験管１０内部の培養液は試験管の長手方向を往復するので培養液が攪拌され、ガス交換が促進される。本発明の試験管１０の貫通孔１３は、試験管本体１２の側壁に設けられる。培養液が長手方向に往復するときには、貫通孔が試験管本体の側壁に設けられる場合の方が貫通孔がキャップに設けられる場合に比べて長期培養における培養液の漏出が起こりにくい。   The rotation axis of the rotator 21 of the rotary culture device 20 of the present invention may be in any direction between the horizontal direction and the vertical direction. The longitudinal axis of the test tube 10 and the axis of the rotator 21 may be parallel, intersect, or may not belong to the same plane. Preferably, the test tube 10 of the present invention is mounted on a rotating culture apparatus including a rotator having a horizontal rotating shaft as shown in FIG. In this case, as the rotator rotates, the culture solution inside the test tube 10 moves so as to be in contact with the entire side wall of the test tube body. When the longitudinal axis of the test tube 10 of the present invention intersects the rotation axis of the rotator or does not belong to the same plane, the culture solution inside the test tube 10 changes the longitudinal direction of the test tube as the rotator rotates. The reciprocating operation stirs the culture solution and promotes gas exchange. The through hole 13 of the test tube 10 of the present invention is provided on the side wall of the test tube body 12. When the culture solution reciprocates in the longitudinal direction, leakage of the culture solution in the long-term culture is less likely when the through hole is provided in the side wall of the test tube body than when the through hole is provided in the cap.

本発明の回転培養システム６０では、複数個の気密容器３０が単一の温度制御装置５０に収容される場合がある。この場合には、気密容器ごとに異なるガス組成及び／又は圧力を設定することが可能になる。   In the rotary culture system 60 of the present invention, a plurality of airtight containers 30 may be accommodated in a single temperature controller 50. In this case, different gas compositions and / or pressures can be set for each hermetic container.

本発明の回転培養システム６０は、試験管１０及び気密容器３０がプラスチックでできており、回転培養装置が電源としての電池２３を含み、ガス組成及び／又は圧力の制御装置４０がガス発生剤又はガス吸収剤の場合がある。この場合には、ガスボンベや圧力調整器のような高額の設備がなくても、ヒト正常細胞の３次元高密度培養を行うことが可能になる。そこで、本発明の回転培養システムは、施設及び予算の制約が厳しい中小病院や診療所における再生医療の普及を促進するのに役立つ。   In the rotary culture system 60 of the present invention, the test tube 10 and the airtight container 30 are made of plastic, the rotary culture device includes a battery 23 as a power source, and the gas composition and / or pressure control device 40 is a gas generating agent or It may be a gas absorbent. In this case, it is possible to perform three-dimensional high-density culture of normal human cells without expensive equipment such as gas cylinders and pressure regulators. Therefore, the rotary culture system of the present invention is useful for promoting the spread of regenerative medicine in small and medium-sized hospitals and clinics where facilities and budget constraints are severe.

本発明の試験管の透視図。The perspective view of the test tube of this invention. 本発明の回転培養装置の透視図。The perspective view of the rotation culture apparatus of this invention. 本発明の回転培養システムの断面模式図。The cross-sectional schematic diagram of the rotation culture system of this invention.

符号の説明Explanation of symbols

１０ 試験管
１１ キャップ
１２ 試験管本体
１３ 貫通孔
１４ 膜
２０ 回転培養装置
２１ ローテータ
２２ モータ
２３ 電池
３０ 気密装置
４０ ガス組成及び／又は圧力の制御装置
５０ 温度制御装置
６０ 回転培養システム DESCRIPTION OF SYMBOLS 10 Test tube 11 Cap 12 Test tube main body 13 Through-hole 14 Membrane 20 Rotating culture device 21 Rotator 22 Motor 23 Battery 30 Airtight device 40 Gas composition and / or pressure control device 50 Temperature control device 60 Rotating culture system

Claims (5)

キャップ付の試験管であって、該試験管の側壁には１個又は２個以上の貫通孔が設けられ、該貫通孔は、通気性があり、かつ、透水性のない膜で閉塞されていることを特徴とする試験管。   A test tube with a cap, wherein one or two or more through holes are provided in a side wall of the test tube, and the through holes are closed with a membrane having air permeability and no water permeability. A test tube characterized by １本又は２本以上の請求項１に記載の試験管を装架することができるローテータと、該ローテータを駆動するモータとを含むことを特徴とする回転培養装置。   A rotating culture apparatus comprising: a rotator capable of mounting one or more test tubes according to claim 1; and a motor for driving the rotator. 電源としての電池を含むことを特徴とする請求項２に記載の回転培養装置。   The rotating culture apparatus according to claim 2, comprising a battery as a power source. 請求項１に記載の試験管と、請求項２又は３に記載の回転培養装置と、前記試験管及び回転培養装置を収容する気密容器と、該気密容器内のガス組成及び／又は圧力の制御装置と、前記気密容器内温度の制御装置とを含むことを特徴とする回転培養システム。   A test tube according to claim 1, a rotary culture device according to claim 2 or 3, an airtight container containing the test tube and the rotary culture device, and control of gas composition and / or pressure in the airtight container. A rotary culture system comprising an apparatus and a controller for controlling the temperature in the hermetic container. 前記試験管及び気密容器はプラスチックでできており、前記ガス組成及び圧力の制御装置はガス発生剤又はガス吸収剤であることを特徴とする請求項４に記載の回転培養システム。   The rotary culture system according to claim 4, wherein the test tube and the airtight container are made of plastic, and the gas composition and pressure control device is a gas generating agent or a gas absorbing agent.
JP2007298869A 2007-11-19 2007-11-19 Compact rotary culture system and culture vessel Withdrawn JP2009118820A (en)

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US9574166B2 (en) 2010-08-18 2017-02-21 Drugmode Aps Bioreactor system
JP2012117910A (en) * 2010-11-30 2012-06-21 Waseda Univ Experiment system and experiment method for evaluating blood adaptability or the like, and sample container to be used for the same
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