JP2007300870A - Tool for holding cultured cell - Google Patents

Tool for holding cultured cell Download PDF

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JP2007300870A
JP2007300870A JP2006133760A JP2006133760A JP2007300870A JP 2007300870 A JP2007300870 A JP 2007300870A JP 2006133760 A JP2006133760 A JP 2006133760A JP 2006133760 A JP2006133760 A JP 2006133760A JP 2007300870 A JP2007300870 A JP 2007300870A
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main body
cultured cells
body part
cultured
cultured cell
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JP4721951B2 (en
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Satoshi Amano
聡 天野
Arinori Ogura
有紀 小倉
Toshio Nishiyama
敏夫 西山
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Shiseido Co Ltd
Tokyo University of Agriculture and Technology NUC
Tokyo University of Agriculture
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Shiseido Co Ltd
Tokyo University of Agriculture and Technology NUC
Tokyo University of Agriculture
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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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings

Abstract

<P>PROBLEM TO BE SOLVED: To efficiently provide elongation stimulation/compression stimulation without shifting cultured cells from an installation surface by firmly holding the cultured cells on the installation surface of a silicon chamber. <P>SOLUTION: The tool for holding cultured cells has an extending and compressing main body part and a holding means for fixing cultured cells to the main body part and elongates/compresses cultured cells according to the extension and compression of the main body part. The main body part is composed of meshes made of a metal, preferably stainless steel and each of the meshes comprises a quadrangle having diagonals in parallel with the length direction and the width direction of the main body part. Consequently, the meshes are extended and compressed in each direction and extended and compressed cultured cells are observed from between the meshes. The holding means is a means which is placed on a plane at the center of the main body part and prevents the cultured cells placed on the plane from sliding when the main body part is extended and compressed. The holding means is preferably a plurality of latching needles and arranged on lines forming the quadrangles. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

本発明は、培養細胞、例えば、培養皮膚または人工皮膚、平滑筋細胞およびコラーゲンゲルを組み合わせた血管内膜モデル（または、この上に内皮細胞を組み合わせた血管モデル）、腱細胞およびコラーゲンを組み合わせた培養腱モデル、骨芽細胞およびコラーゲンゲルを組み合わせた骨形成モデル、あるいは、生体由来の皮膚や血管や腱そのものの器官培養等の、伸展・圧縮装置に用いられる培養細胞保持器具に関する。   The present invention combines cultured cells, for example, cultured or artificial skin, an endometrial model combining smooth muscle cells and collagen gel (or a vascular model combining endothelial cells thereon), tendon cells and collagen. The present invention relates to a cultured cell holding device used in an extension / compression device, such as a cultured tendon model, a bone formation model combining osteoblasts and collagen gel, or organ culture of living body-derived skin, blood vessels, or tendons themselves.

培養皮膚等の培養細胞に伸展刺激・圧縮刺激を負荷し、そのような機械的刺激を受けた細胞におけるシグナルトランスダクション、タンパク質発現、遺伝子発現の変化の研究や、形態観察（蛍光抗体法）等を行うために、培養細胞の伸展・圧縮装置が用いられている（非特許文献１、非特許文献２）。   Cell culture cells such as cultured skin are subjected to stretching and compression stimuli, and signal transduction, protein expression, and changes in gene expression in cells that have undergone such mechanical stimulation, morphological observation (fluorescent antibody method), etc. In order to perform this, a stretched / compressed apparatus for cultured cells is used (Non-Patent Document 1, Non-Patent Document 2).

対象の培養細胞をこのような装置に据え付けるために、略透明のシリコン薄膜の据置面を有し、全体がシリコンでできているシリコンチャンバーを用いる手段がある。これは据置面に培養細胞を置き、据置面と培養細胞との間の静止摩擦力のみによって伸展・圧縮時に滑らないようにされている。   In order to install the target cultured cells in such an apparatus, there is a means using a silicon chamber having a substantially transparent silicon thin-film mounting surface and made entirely of silicon. In this method, the cultured cells are placed on the stationary surface, and only the static frictional force between the stationary surface and the cultured cells is prevented from slipping during extension / compression.

Experimental Cell Reserch 247, 320-328(1999)Experimental Cell Reserch 247, 320-328 (1999) THE JOURNAL OF INVESTIGATIVE DERMATOLOGY, vol 97 No.5 Novem-ber 1991, P898-P902THE JOURNAL OF INVESTIGATIVE DERMATOLOGY, vol 97 No.5 Novem-ber 1991, P898-P902

しかし、この方法によると、より大きい刺激を負荷するためにより大きな伸縮の変位を与えた場合、静止摩擦力を超えて培養細胞が据置面に対してずれてしまい、正確な観察等ができないという問題点がある。   However, according to this method, when a larger expansion / contraction displacement is applied to load a larger stimulus, the cultured cells are displaced with respect to the stationary surface beyond the static friction force, so that accurate observation or the like cannot be performed. There is a point.

前記課題を解決するために１番目の発明によれば、伸縮する本体部と本体部に培養細胞を固定する保持手段とを有し、本体部の伸縮に応じて培養細胞を伸展・圧縮する。これによって、伸展刺激・圧縮刺激負荷時に培養細胞がずれないようにしっかりと保持できるようになる。   In order to solve the above-mentioned problem, according to the first invention, it has a main body part that expands and contracts and a holding means that fixes cultured cells to the main body part, and expands and compresses cultured cells according to the expansion and contraction of the main body part. This makes it possible to hold the cultured cells firmly so that they do not shift when subjected to extension stimulation or compression stimulation.

また、２番目の発明によれば１番目の発明において、本体部が金属製メッシュから成る。   According to a second aspect, in the first aspect, the main body is made of a metal mesh.

また、３番目の発明によれば２番目の発明において、金属製メッシュが本体部の長手方向と短手方向に平行に各対角線を有する四角形から成る。これによって、本体部が各方向に伸縮可能となると共に、メッシュの間から培養細胞を観察することが可能となる。   According to the third invention, in the second invention, the metal mesh is formed of a quadrangle having diagonal lines parallel to the longitudinal direction and the short direction of the main body. Thereby, the main body can be expanded and contracted in each direction, and the cultured cells can be observed from between the meshes.

また、４番目の発明によれば１番目から３番目のいずれかの発明において、保持手段が本体部と同じ材質から成る。   According to the fourth invention, in any one of the first to third inventions, the holding means is made of the same material as the main body.

また、５番目の発明によれば１番目から４番目のいずれかの発明において、保持手段が培養細胞を本体部に係止する複数の係止針である。これによって、培養細胞を突き刺して保持することが可能となる。   According to the fifth invention, in any one of the first to fourth inventions, the holding means is a plurality of locking needles for locking the cultured cell to the main body. This makes it possible to pierce and hold the cultured cells.

また、６番目の発明によれば５番目の発明において、保持手段が四角形を形成する線上に配置されている。   According to the sixth aspect, in the fifth aspect, the holding means is arranged on a line forming a quadrangle.

また、７番目の発明によれば１番目から６番目のいずれかの発明において、本体部がシリコンチャンバーを介して培養細胞の伸展・圧縮装置に取付可能な形状である。これによって、本発明による保持器具を公知のシリコンチャンバーに固定クリップを用いて取り付けることが可能となる。   According to a seventh invention, in any one of the first to sixth inventions, the main body has a shape that can be attached to a cell extension / compression device via a silicon chamber. As a result, the holding device according to the present invention can be attached to a known silicon chamber using a fixing clip.

培養細胞をシリコンチャンバーの据置面上にしっかりと保持できることによって、培養細胞が据置面からずれることなく伸展刺激・圧縮刺激を効率的に負荷することが可能となる。   Since the cultured cells can be firmly held on the stationary surface of the silicon chamber, the cultured cells can be efficiently loaded with extension and compression stimuli without being displaced from the stationary surface.

図１は本発明による培養細胞の保持器具１の斜視図を示し、図２はその上面図、図３はその側面図を示す。   FIG. 1 is a perspective view of a cultured cell holding device 1 according to the present invention, FIG. 2 is a top view thereof, and FIG. 3 is a side view thereof.

ここで培養細胞には、培養皮膚または人工皮膚のみならず、例えば、平滑筋細胞およびコラーゲンゲルを組み合わせた血管内膜モデル（または、この上に内皮細胞を組み合わせた血管モデル）、腱細胞およびコラーゲンを組み合わせた培養腱モデル、骨芽細胞およびコラーゲンゲルを組み合わせた骨形成モデル、あるいは、生体由来の皮膚や血管や腱そのものの器官培養等を含む。   Here, the cultured cells include not only cultured skin or artificial skin, but also, for example, an intima model in which smooth muscle cells and collagen gel are combined (or a blood vessel model in which endothelial cells are combined thereon), tendon cells and collagen. Cultured bone tendon model combining osteoblasts and collagen gel, or organ culture of living body-derived skin, blood vessels and tendons themselves.

保持器具１は本体部２と保持手段３とを有している。本体部２は、好ましくはステンレスである金属製のメッシュからできており、個々のメッシュは本体部２の長手方向と短手方向に平行に各対角線を有する四角形から成る。そのため、各方向に本体部２を引っ張ると各対角線の長さ比が変化することによって、それぞれの方向に伸縮可能となると共に、メッシュの間からは伸縮された培養細胞を観察等することが可能となる。本体部２の中央は平面となっており、両端面は後述のシリコンチャンバー５に嵌合するように屈曲部４を形成している。   The holding device 1 has a main body 2 and holding means 3. The main body 2 is made of a metal mesh, preferably stainless steel, and each mesh is made of a quadrangle having diagonal lines parallel to the longitudinal direction and the short direction of the main body 2. Therefore, when the main body 2 is pulled in each direction, the length ratio of each diagonal line changes, so that it is possible to expand and contract in each direction and observe the expanded and contracted cultured cells from between the meshes. It becomes. The center of the main body 2 is a flat surface, and both end surfaces are formed with bent portions 4 so as to be fitted into a silicon chamber 5 described later.

一方、保持手段３は、本体部２の中央の平面上に配置され、そこに置かれる培養細胞が本体部２を伸縮した際に滑らぬように係止するための手段である。好ましくは、複数の係止針であり、本体部１と同じ材質から成る。図に示す実施形態において、保持手段３が四角形を形成する線上に配置されているが、円形等、その他の形状であってもよい。   On the other hand, the holding means 3 is arranged on a central plane of the main body portion 2 and is a means for locking so that the cultured cells placed thereon do not slide when the main body portion 2 expands and contracts. Preferably, the plurality of locking needles are made of the same material as the main body 1. In the embodiment shown in the figure, the holding means 3 is arranged on a line forming a quadrangle, but may be other shapes such as a circle.

図４はシリコンでできた公知のシリコンチャンバー５の斜視図である。シリコンチャンバー５は、図の左右の側壁７と、図の左右方向に伸縮し易いように側壁７に比べて薄く形成された側壁８と、培養細胞を観察し易いように略透明のシリコン薄膜でできた底を塞ぐ据置面６とでチャンバーを形成している。さらに、左右の側壁７には、培養細胞の伸展・圧縮装置の固定ピン（図示略）への取り付け用に鉛直に貫通孔９が設けられている。   FIG. 4 is a perspective view of a known silicon chamber 5 made of silicon. The silicon chamber 5 is composed of a side wall 7 on the left and right side of the figure, a side wall 8 formed thinner than the side wall 7 so as to easily expand and contract in the left and right direction in the figure, and a substantially transparent silicon thin film so that cultured cells can be easily observed. A chamber is formed by the stationary surface 6 that closes the completed bottom. Further, the left and right side walls 7 are provided with through-holes 9 vertically for attachment to a fixing pin (not shown) of a cell expansion / compression device.

図５は、本発明による保持器具１をシリコンチャンバー５に固定クリップ１０を用いて取り付けた状態の上面図であり、線Ａ−Ａにおける断面図を図５に示す。   FIG. 5 is a top view of the state in which the holding device 1 according to the present invention is attached to the silicon chamber 5 using the fixing clip 10, and a cross-sectional view taken along line AA is shown in FIG. 5.

保持器具１の各屈曲部４をシリコンチャンバー５の対応する側壁７に嵌るように被せ、さらに、好ましくはステンレス製の固定クリップ１０によって、その弾性を利用して、保持器具１がシリコンチャンバー５から外れたりずれたりしないように挟んで止めている。この状態で例えばコラーゲンゲルのような培養細胞１１を保持手段３上に突き刺すようにセットし、場合によっては培養液等でチャンバーを満たす。   Each bent portion 4 of the holding device 1 is covered with a corresponding side wall 7 of the silicon chamber 5, and preferably, the holding device 1 is removed from the silicon chamber 5 by utilizing the elasticity of a fixing clip 10 made of stainless steel. It is clamped so that it does not come off or shift. In this state, a cultured cell 11 such as a collagen gel is set so as to pierce the holding means 3, and the chamber is filled with a culture solution or the like in some cases.

これらを培養細胞の伸展・圧縮装置の固定ピンを貫通孔９に通して取り付けると、伸展刺激・圧縮刺激を負荷するためのシリコンチャンバーの伸縮に応じて培養細胞も伸縮し、大きな変位を与えても培養細胞１１がずれてしまうことなく、観察等を行うことが可能となる。   When these are attached through the through-hole 9 with the fixing pins of the cell expansion / compression device, the cell expansion / contraction in response to the expansion / contraction of the silicon chamber for loading the expansion / compression stimulus causes a large displacement. In addition, observation or the like can be performed without the cultured cells 11 being displaced.

前述の実施形態において、刺激負荷は一軸方向であったが、メッシュの構造から明らかなように二軸方向の刺激負荷に適用することも可能である。   In the above-described embodiment, the stimulation load is uniaxial. However, as is apparent from the mesh structure, the stimulation load can be applied to the biaxial stimulation load.

本発明による培養細胞の保持器具１の斜視図である。It is a perspective view of the holding device 1 of the cultured cell by this invention. 図１に示された保持器具１の上面図である。FIG. 2 is a top view of the holding device 1 shown in FIG. 1. 図１に示された保持器具１の側面図である。It is a side view of the holding fixture 1 shown by FIG. 公知のシリコンチャンバー５の斜視図である。2 is a perspective view of a known silicon chamber 5. FIG. 本発明による保持器具１をシリコンチャンバー５に固定クリップ１０を用いて取り付けた状態での上面図である。It is a top view in the state where holding fixture 1 by the present invention was attached to silicon chamber 5 using fixed clip 10. FIG. 図５の線Ａ−Ａにおける断面図である。It is sectional drawing in line AA of FIG.

符号の説明Explanation of symbols

１ 保持器具
２ 本体部
３ 保持手段
４ 屈曲部
５ シリコンチャンバー
６ 据置面
７、８ 側壁
９ 貫通孔
１０ 固定クリップ
１１ 培養細胞 DESCRIPTION OF SYMBOLS 1 Holding tool 2 Main-body part 3 Holding means 4 Bending part 5 Silicon chamber 6 Stationary surface 7, 8 Side wall 9 Through-hole 10 Fixed clip 11 Cultured cell

Claims (7)

伸縮する本体部と該本体部に培養細胞を固定する保持手段とを有し、前記本体部の伸縮に応じて前記培養細胞を伸展・圧縮する培養細胞保持器具。   A cultured cell holding device that has a main body portion that expands and contracts and a holding means that fixes cultured cells to the main body portion, and that expands and compresses the cultured cells according to the expansion and contraction of the main body portion. 前記本体部が金属製メッシュから成る請求項１に記載の培養細胞保持器具。   The cultured cell holding device according to claim 1, wherein the main body portion is made of a metal mesh. 前記金属製メッシュが前記本体部の長手方向と短手方向に各対角線を有する四角形から成る請求項２に記載の培養細胞保持器具。   The cultured cell holding device according to claim 2, wherein the metal mesh is formed of a quadrangle having diagonal lines in the longitudinal direction and the short direction of the main body. 前記保持手段が前記本体部と同じ材質から成る請求項１から３のいずれかに記載の培養細胞保持器具。   The cultured cell holding device according to any one of claims 1 to 3, wherein the holding means is made of the same material as the main body. 前記保持手段が前記培養細胞を前記本体部に係止する複数の係止針である請求項１から４のいずれかに記載の培養細胞保持器具。   The cultured cell holding device according to any one of claims 1 to 4, wherein the holding means is a plurality of locking needles for locking the cultured cell to the main body. 前記保持手段が四角形を形成する線上に配置されている請求項５に記載の培養細胞保持器具。   The cultured cell holding device according to claim 5, wherein the holding means is arranged on a line forming a quadrangle. 前記本体部がシリコンチャンバーを介して培養細胞の伸展・圧縮装置に取付可能な形状である請求項１から６のいずれかに記載の培養細胞保持器具。   The cultured cell holding device according to any one of claims 1 to 6, wherein the main body has a shape that can be attached to an apparatus for extending and compressing cultured cells via a silicon chamber.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008123614A1 (en) * 2007-03-30 2008-10-16 Kyushu University, National University Corporation Method for production of three-dimensional structure of cells
JP2011162507A (en) * 2010-02-12 2011-08-25 Pias Arise Kk Inhibitor of extension/stimulation-mediated production of collagen decomposition enzyme
JP2015070824A (en) * 2013-10-04 2015-04-16 株式会社椿本チエイン Tissue peeling prevention plate and culture unit

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005087913A1 (en) * 2004-03-11 2005-09-22 Nagoya Industrial Science Research Institute Culture device

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005087913A1 (en) * 2004-03-11 2005-09-22 Nagoya Industrial Science Research Institute Culture device

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008123614A1 (en) * 2007-03-30 2008-10-16 Kyushu University, National University Corporation Method for production of three-dimensional structure of cells
US8198086B2 (en) 2007-03-30 2012-06-12 Kyushu University, National University Corporation Method for production of three-dimensional structure of cells
JP2011162507A (en) * 2010-02-12 2011-08-25 Pias Arise Kk Inhibitor of extension/stimulation-mediated production of collagen decomposition enzyme
JP2015070824A (en) * 2013-10-04 2015-04-16 株式会社椿本チエイン Tissue peeling prevention plate and culture unit

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