JPH04110500U - Cell culture container - Google Patents
Cell culture containerInfo
- Publication number
- JPH04110500U JPH04110500U JP2219291U JP2219291U JPH04110500U JP H04110500 U JPH04110500 U JP H04110500U JP 2219291 U JP2219291 U JP 2219291U JP 2219291 U JP2219291 U JP 2219291U JP H04110500 U JPH04110500 U JP H04110500U
- Authority
- JP
- Japan
- Prior art keywords
- culture
- cells
- tank support
- semipermeable membrane
- 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.)
- Withdrawn
Links
- 238000004113 cell culture Methods 0.000 title claims abstract description 18
- 239000012528 membrane Substances 0.000 claims abstract description 30
- 230000002093 peripheral effect Effects 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 description 41
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 25
- 229910052760 oxygen Inorganic materials 0.000 description 25
- 239000001301 oxygen Substances 0.000 description 25
- 239000001963 growth medium Substances 0.000 description 13
- 210000004408 hybridoma Anatomy 0.000 description 8
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- 210000004748 cultured cell Anatomy 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 206010035226 Plasma cell myeloma Diseases 0.000 description 3
- 239000004809 Teflon Substances 0.000 description 3
- 229920006362 Teflon® Polymers 0.000 description 3
- 210000004102 animal cell Anatomy 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 description 3
- 239000001569 carbon dioxide Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 201000000050 myeloid neoplasm Diseases 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
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- 230000001605 fetal effect Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 210000003292 kidney cell Anatomy 0.000 description 2
- 210000005265 lung cell Anatomy 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000010935 stainless steel Substances 0.000 description 2
- 229960000187 tissue plasminogen activator Drugs 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 2
- 239000004925 Acrylic resin Substances 0.000 description 1
- 229920000178 Acrylic resin Polymers 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
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- 241000282552 Chlorocebus aethiops Species 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- 208000007913 Pituitary Neoplasms Diseases 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 208000024447 adrenal gland neoplasm Diseases 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229920001973 fluoroelastomer Polymers 0.000 description 1
- 210000003953 foreskin Anatomy 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000003810 lymphokine-activated killer cell Anatomy 0.000 description 1
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- 229920002239 polyacrylonitrile Polymers 0.000 description 1
- 229920005668 polycarbonate resin Polymers 0.000 description 1
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- 229920000728 polyester Polymers 0.000 description 1
- 229920013716 polyethylene resin Polymers 0.000 description 1
- -1 polypropylene Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 229920005990 polystyrene resin Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 239000004627 regenerated cellulose Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 239000004945 silicone rubber Substances 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
Abstract
(57)【要約】
【目的】 培養を継続しながら連続的または間けつ的な
培養液の交換をコンパクトな構成で行うことができ、組
立が容易な細胞培養用容器を提供することにある。
【構成】 本考案の細胞培養用容器は、周側面に窓部が
形成されてなる筒状の槽支持体および当該槽支持体の窓
部を塞ぐよう設けられた筒状の半透膜部材よりなる培養
槽本体と、当該培養槽本体が格納された、開口部を有す
る外筒容器と、当該外筒容器の開口部を気密に塞ぐ蓋部
材とにより構成され、前記槽支持体は、蓋部材の内面に
形成された嵌合部と嵌合して支持され、前記半透膜部材
は、その一端が前記嵌合部および前記槽支持体の周側面
によって液密に挟持されていることを特徴とする。
(57) [Summary] [Purpose] The purpose is to provide a cell culture container that allows continuous or intermittent culture solution exchange while continuing culture with a compact configuration and is easy to assemble. [Structure] The cell culture container of the present invention consists of a cylindrical tank support with a window formed on the circumferential side and a cylindrical semipermeable membrane member provided to close the window of the tank support. The tank support is composed of a culture tank main body, an outer cylindrical container having an opening in which the culture tank main body is stored, and a lid member that airtightly closes the opening of the outer cylindrical container. The semipermeable membrane member is fitted and supported by a fitting part formed on an inner surface of the semipermeable membrane member, and one end of the semipermeable membrane member is fluid-tightly held between the fitting part and the circumferential side of the tank support. shall be.
Description
【０００１】0001
本考案は、微生物、植物細胞および動物細胞（以下これらを単に「細胞等」と いう）、特に動物細胞の培養に好適に用いられる細胞培養用容器に関する。 This invention applies to microorganisms, plant cells, and animal cells (hereinafter referred to simply as "cells, etc."). ), particularly relates to a cell culture container suitably used for culturing animal cells.
【０００２】0002
細胞等の培養を高い効率で実行するためには、培養される細胞等を常に新しい 培養液と接触させることが肝要であり、従って培養用容器内に常に新しい培養液 を供給することが必要である。また、培養される細胞に十分な酸素が供給される ことも重要である。 In order to culture cells etc. with high efficiency, it is necessary to always use fresh cells, etc. to be cultured. Contact with the culture medium is essential, so there is always fresh culture medium in the culture container. It is necessary to supply It also provides sufficient oxygen to the cells being cultured. It is also important that
【０００３】 しかして、従来においては、一部が半透膜で構成された容器本体と、この容器 本体が格納された外筒容器と、酸素供給機構とを有してなる細胞培養用容器を用 いることにより、半透膜を利用した培養液の供給および酸素供給機構による酸素 の供給が行われていた。0003 However, in the past, the container body was partially composed of a semipermeable membrane, and the container A cell culture container is used, which has an outer cylindrical container housing the main body and an oxygen supply mechanism. By using a semi-permeable membrane to supply culture medium and an oxygen supply mechanism, were being supplied.
【０００４】0004
しかしながら、従来の細胞培養用容器においては、半透膜の支持や酸素供給機 構の構成などにおいて多大の問題があり、コンパクトなものは提供されていない 。 However, in conventional cell culture containers, support for semipermeable membranes and oxygen supply devices are insufficient. There are many problems with the structure, and a compact version is not available. .
【０００５】 本考案は、以上のような事情に基づいてなされたものであって、その目的は、 培養を継続しながら連続的または間けつ的な培養液の交換をコンパクトな構成で 行うことができ、組立が容易な細胞培養用容器を提供することにある。[0005] This invention was made based on the above circumstances, and its purpose is to: A compact configuration allows for continuous or intermittent culture medium exchange while continuing culture. An object of the present invention is to provide a container for cell culture that can be easily assembled.
【０００６】[0006]
本考案の細胞培養用容器は、周側面に窓部が形成されてなる筒状の槽支持体お よび当該槽支持体の窓部を塞ぐよう設けられた筒状の半透膜部材よりなる培養槽 本体と、当該培養槽本体が格納された、開口部を有する外筒容器と、当該外筒容 器の開口部を気密に塞ぐ蓋部材とにより構成され、前記槽支持体は、蓋部材の内 面に形成された嵌合部と嵌合して支持され、前記半透膜部材は、その一端が前記 嵌合部および前記槽支持体の周側面によって液密に挟持されていることを特徴と する。 The cell culture container of the present invention is a cylindrical tank support with a window formed on the peripheral side. and a cylindrical semipermeable membrane member provided to close the window of the tank support. A main body, an outer cylindrical container having an opening in which the culture tank main body is stored, and the outer cylindrical container. and a lid member that airtightly closes the opening of the vessel, and the tank support is arranged inside the lid member. The semipermeable membrane member is supported by being fitted into a fitting part formed on the surface, and one end of the semipermeable membrane member is fitted with the fitting part formed on the surface. It is characterized by being fluid-tightly held between the fitting part and the circumferential side of the tank support. do.
【０００７】[0007]
以下本考案の実施例を図面に基づいて説明する。 図１は、本考案の細胞培養用容器の一実施例を示す説明用断面図である。 この実施例の細胞培養用容器は、培養槽本体１と、培養槽本体１が格納された 、開口部21を有する外筒容器２と、外筒容器２の開口部21を気密に塞ぐ蓋部材３ と、螺旋状またはうず巻状に巻かれた状態で培養槽本体１内に設けられた酸素供 給管４とにより構成されている。 Embodiments of the present invention will be described below based on the drawings. FIG. 1 is an explanatory cross-sectional view showing one embodiment of the cell culture container of the present invention. The cell culture container of this example includes a culture tank main body 1 and a culture tank main body 1 stored therein. , an outer cylindrical container 2 having an opening 21, and a lid member 3 that airtightly closes the opening 21 of the outer cylindrical container 2. and an oxygen supply provided inside the culture tank body 1 in a spiral or spirally wound state. It is composed of a supply pipe 4.
【０００８】 培養槽本体１は、周側面に窓部12が形成されてなる円筒状の槽支持体11、槽支 持体11の窓部12を塞ぐよう設けられた円筒状の半透膜部材13および円柱状の底板 部材15よりなる。[0008] The culture tank body 1 includes a cylindrical tank support 11 having a window 12 formed on the circumferential side, and a tank support. A cylindrical semipermeable membrane member 13 and a cylindrical bottom plate provided to close the window 12 of the support body 11 Consists of member 15.
【０００９】 培養槽本体は、その内容積が100 〜5000ミリリットル、特に 100〜3000ミリリ ットルであるものが好ましい。[0009] The culture tank itself has an internal volume of 100 to 5000 ml, especially 100 to 3000 ml. Preferably, the
【００１０】 培養槽本体の形状は円筒状が好ましいが、これに限定されるものではなく、立 方体、直方体などであってもよい。培養槽本体の形状が円筒状である場合におい て、培養内容積（Ｖml）に対する円筒内径（Ｌcm）の比（Ｌ／Ｖ）は、表１に示 す範囲のものが好ましい。この表１をグラフ化して図４に示す。 図４において、Ａは好ましい範囲を示し、Ｂは特に好ましい範囲を示す。0010 The shape of the culture tank body is preferably cylindrical, but is not limited to this. It may be a parallelepiped, a rectangular parallelepiped, or the like. If the culture tank body is cylindrical, odor may occur. The ratio (L/V) of the cylinder inner diameter (Lcm) to the culture internal volume (Vml) is shown in Table 1. Preferably, the range is within this range. Table 1 is graphed and shown in FIG. 4. In FIG. 4, A indicates a preferred range, and B indicates a particularly preferred range.
【００１１】[0011]
【表１】 [Table 1]
【００１２】 培養槽本体１を構成する槽支持体の材質としては、例えば硬質ガラス、石英ガ ラス、ステンレス（ＳＵＳ３１６）、ポリプロピレン樹脂、ポリエチレン樹脂、 ポリスチレン樹脂、ポリカーボネート樹脂、アクリル樹脂、テフロン樹脂などを 挙げることができる。0012 The material of the tank support that constitutes the culture tank body 1 may be, for example, hard glass or quartz glass. lath, stainless steel (SUS316), polypropylene resin, polyethylene resin, Polystyrene resin, polycarbonate resin, acrylic resin, Teflon resin, etc. can be mentioned.
【００１３】 培養槽本体１を構成する円筒状の半透膜部材13は、通常、10μｍ以下の孔径を 有する膜よりなる。これにより、培養液は半透膜部材13を透過できるが、細胞は 透過することができない。[0013] The cylindrical semipermeable membrane member 13 that constitutes the culture tank body 1 usually has a pore diameter of 10 μm or less. It consists of a membrane with This allows the culture solution to pass through the semipermeable membrane member 13, but the cells cannot be penetrated.
【００１４】 半透膜部材13の材質としては、例えば再生セルロース、セルロースアセテート などのセルロース系の半透膜、およびポリエステル、ポリアクリロニトリルなど の合成高分子化合物よりなる半透膜を挙げることができる。[0014] Examples of the material of the semipermeable membrane member 13 include regenerated cellulose and cellulose acetate. Cellulose-based semipermeable membranes such as polyester, polyacrylonitrile, etc. Examples include semipermeable membranes made of synthetic polymer compounds.
【００１５】 本実施例において、半透膜部材13は、槽支持体11に形成された窓部12を外側か ら塞ぐよう槽支持体11の外周側面に当接されている。[0015] In this embodiment, the semipermeable membrane member 13 is arranged so that the window 12 formed in the tank support 11 faces outward. It is brought into contact with the outer circumferential side surface of the tank support 11 so as to close it.
【００１６】 底板部材15の径15Ｄは、槽支持体11の内径11ｄより僅かに小さいがほぼ等しい 大きさであり、これにより底板部材15は槽支持体11に嵌合されている。そして半 透膜部材13の下端13Ｂは、槽支持体11の下端11Ｂにおいて折り返された状態で槽 支持体11の内周側面および底板部材15の外周面に液密に挟持されている。[0016] The diameter 15D of the bottom plate member 15 is slightly smaller than the inner diameter 11d of the tank support 11, but approximately equal to it. Due to this size, the bottom plate member 15 is fitted into the tank support 11. and half The lower end 13B of the membrane member 13 is folded back at the lower end 11B of the tank support 11 and It is fluid-tightly sandwiched between the inner circumferential side surface of the support body 11 and the outer circumferential surface of the bottom plate member 15.
【００１７】 外筒容器２の開口部21のフランジ外周面および蓋部材３の外周面には、それぞ れ同一ピッチの螺合部21Ａが形成されている。 ５はリング部材であって、リング部材５の内周面には、螺合部21Ａと同一ピッ チの螺合部５Ａが形成されている。これにより、リング部材５を介して外筒容器 ２と蓋部材３とが固定される。外筒容器２の開口部21には、Ｏリング溝23内にお いてシリコンゴムよりなるＯリング６が配置されており、これにより外筒容器２ の開口部21は蓋部材３により気密に塞がれる。[0017] The outer peripheral surface of the flange of the opening 21 of the outer cylindrical container 2 and the outer peripheral surface of the lid member 3 are respectively The threaded portions 21A having the same pitch are formed. 5 is a ring member, and the inner peripheral surface of the ring member 5 has the same pitch as the threaded portion 21A. A threaded portion 5A is formed. As a result, the outer cylindrical container is inserted through the ring member 5. 2 and the lid member 3 are fixed. The opening 21 of the outer cylindrical container 2 has an O-ring groove 23 inside. An O-ring 6 made of silicone rubber is disposed in the outer cylindrical container 2. The opening 21 is hermetically closed by the lid member 3.
【００１８】 蓋部材３の内面36には、円筒状突起よりなる嵌合部37が形成されている。この 嵌合部37の外径37Ｄは、槽支持体11の内径11ｄより僅かに小さいがほぼ等しい大 きさであり、これにより槽支持体11は嵌合部37と嵌合して支持されている。そし て半透膜部材13の上端13Ａは、槽支持体11の上端11Ａにおいて折り返された状態 で槽支持体11の内周側面および嵌合部37の外周面に液密に挟持されている。[0018] A fitting portion 37 made of a cylindrical projection is formed on the inner surface 36 of the lid member 3. this The outer diameter 37D of the fitting part 37 is slightly smaller than the inner diameter 11d of the tank support 11, but is approximately the same size. Due to this, the tank support 11 is fitted into the fitting portion 37 and supported. stop The upper end 13A of the semipermeable membrane member 13 is folded back at the upper end 11A of the tank support 11. It is fluid-tightly sandwiched between the inner circumferential side surface of the tank support 11 and the outer circumferential surface of the fitting part 37.
【００１９】 蓋部材３には、さらに細胞供給口31、酸素供給口32、酸素排出口33、培養液供 給口34、培養液排出口35が形成されている。細胞供給口31は、外部空間と培養槽 本体１の内部空間を連通するものであり、培養槽本体１内への培養液および細胞 の添加、これらの回収、さらに培養中におけるサンプリング操作は、この細胞供 給口31を介して行われる。38は細胞供給口31を塞ぐ止め栓である。[0019] The lid member 3 further includes a cell supply port 31, an oxygen supply port 32, an oxygen discharge port 33, and a culture solution supply port. A supply port 34 and a culture solution discharge port 35 are formed. The cell supply port 31 is connected to the external space and the culture tank. It communicates the internal space of the main body 1, and allows the culture solution and cells to enter the culture tank main body 1. The addition of cells, their collection, and sampling operations during culturing are This is done through the feeding port 31. 38 is a stopper that closes the cell supply port 31.
【００２０】 酸素供給口32および酸素排出口33の各々は、外部口金32ａおよび外部口金33ａ を介して例えば空気ポンプよりなる酸素供給機構（図示省略）に接続され、内部 口金32ｂおよび内部口金33ｂを介して酸素供給管４の端部41および端部42に接続 されている。これにより、培養槽本体１内の細胞に酸素供給機構からの滅菌され た酸素が供給される。[0020] The oxygen supply port 32 and the oxygen discharge port 33 are connected to an external cap 32a and an external cap 33a, respectively. For example, the internal Connected to end 41 and end 42 of oxygen supply pipe 4 via cap 32b and internal cap 33b has been done. This allows the cells in the culture tank body 1 to be sterilized from the oxygen supply mechanism. oxygen is supplied.
【００２１】 培養液供給口34および培養液排出口35の各々は、外部口金34ａおよび外部口金 35ａに接続されたシリコーン製またはテフロン製のチューブ（図示省略）を介し て、例えばポンプよりなる培養液供給機構（図示省略）に接続されている。これ により、新しい培養液は、培養槽本体１の外面、外筒容器２の内面および蓋部材 ３によって区画される培養液循環領域Ｘに連続的または間けつ的に供給され、半 透膜部材13を透過して培養槽本体１内の細胞に供給される。[0021] The culture solution supply port 34 and the culture solution discharge port 35 are connected to an external cap 34a and an external cap, respectively. 35a through a silicone or Teflon tube (not shown). For example, it is connected to a culture solution supply mechanism (not shown) consisting of a pump. this Therefore, the new culture solution is poured onto the outer surface of the culture tank body 1, the inner surface of the outer cylinder container 2, and the lid member. The culture solution is supplied continuously or intermittently to the culture solution circulation area X defined by 3. It passes through the membrane member 13 and is supplied to the cells in the culture tank body 1.
【００２２】 外筒容器２および蓋部材３の材質としては、槽支持体11と同様のガラス、ステ ンレス、樹脂などを挙げることができる。[0022] The material of the outer cylindrical container 2 and the lid member 3 may be the same glass or stainless steel as the tank support 11. Examples include adhesives, resins, etc.
【００２３】 酸素供給管４は、螺旋状またはうず巻状に巻かれた状態で培養槽本体１内の中 央部に設けられいるので、格納スペースを小さくすることができ、細胞への酸素 の供給をコンパクトな構成で行うことができる。[0023] The oxygen supply pipe 4 is wound in a spiral or spiral shape inside the culture tank main body 1. Since it is located in the center, the storage space can be reduced and oxygen to the cells can be reduced. can be supplied with a compact configuration.
【００２４】 酸素供給管４としては、内径0.5 〜2.5 mm、外径1.0 〜4.0 mm、培養容積１リ ットルあたりの長さが５〜30ｍのものを好ましく用いることができ、特に好まし いものとして、内径1.0 mm、外径1.5 mm、培養容積１リットルあたりの長さが10 ｍの酸素供給管を挙げることができる。酸素供給管４の材質としては、シリコン ゴム、テフロン、フッ素ゴムなどを挙げることができる。[0024] The oxygen supply pipe 4 has an inner diameter of 0.5 to 2.5 mm, an outer diameter of 1.0 to 4.0 mm, and a culture volume of 1 liter. Those with a length per liter of 5 to 30 m can be preferably used, and particularly preferred. As a microorganism, the inner diameter is 1.0 mm, the outer diameter is 1.5 mm, and the length is 10 mm per liter of culture volume. m oxygen supply pipes can be mentioned. The material of the oxygen supply pipe 4 is silicon. Examples include rubber, Teflon, and fluororubber.
【００２５】 本実施例によれば、培養槽本体１を構成する槽支持体11が、蓋部材３の嵌合部 37と嵌合して支持され、半透膜部材13の上端13Ａは、槽支持体11の上端11Ａにお いて折り返された状態で槽支持体11の内周側面および嵌合部37の外周面に挟持さ れているので、コンパクトな構成でありながら新しい培養液を培養槽本体１内の 細胞に連続的に供給することができる。 また、半透膜部材13の上端13Ａおよび下端13Ｂが液密に挟持されているので、 培養槽本体１内の細胞が培養液循環領域Ｘへ漏れることはない。 更に、槽支持体11は、嵌合部37に嵌合支持されているので、培養槽本体１を蓋 部材３に容易に着脱することができる。[0025] According to this embodiment, the tank support 11 constituting the culture tank main body 1 is connected to the fitting portion of the lid member 3. 37 and is supported, and the upper end 13A of the semipermeable membrane member 13 is connected to the upper end 11A of the tank support 11. It is held between the inner circumferential surface of the tank support 11 and the outer circumferential surface of the fitting part 37 in the folded state. Because it has a compact structure, new culture solution can be stored in the culture tank main body 1. It can be fed continuously to cells. In addition, since the upper end 13A and lower end 13B of the semipermeable membrane member 13 are sandwiched in a liquid-tight manner, Cells within the culture tank body 1 will not leak into the culture solution circulation area X. Furthermore, since the tank support 11 is fitted and supported by the fitting part 37, the culture tank body 1 can be closed with a lid. It can be easily attached to and detached from the member 3.
【００２６】 本実施例の細胞培養用容器は、コンパクトな構成でありながら、細胞への酸素 の供給および連続的または間けつ的な培養液の交換を行うことができ、高い密度 で細胞を培養することができ、その組立も容易である。[0026] The cell culture container of this example has a compact structure, yet provides oxygen to the cells. supply and continuous or intermittent culture medium exchange, high density Cells can be cultured in this device, and it is easy to assemble.
【００２７】 なお、本考案においては、上記の実施例に限定されるものではなく、種々の変 更が可能である。 例えば、図２に示すように、半透膜部材13が、槽支持体11に形成された窓部12 を内側から塞ぐよう槽支持体11の内周側面に当接されていてもよい。[0027] Note that the present invention is not limited to the above embodiments, and various modifications may be made. Changes are possible. For example, as shown in FIG. It may be brought into contact with the inner circumferential side surface of the tank support 11 so as to close it from the inside.
【００２８】 また、図３に示すように、槽支持体と底板部材とが一体になっていてもよく、 この場合においては、袋状の半透膜部材13' を用い、その上端13’Ａのみが、槽 支持体11’の上端11’Ａにおいて折り返された状態で槽支持体11’の内周側面お よび蓋部材３の嵌合部37の外周面に挟持されていてもよい。[0028] Further, as shown in FIG. 3, the tank support and the bottom plate member may be integrated, In this case, a bag-shaped semipermeable membrane member 13' is used, and only its upper end 13'A is connected to the tank. The inner peripheral side surface of the tank support 11' is folded back at the upper end 11'A of the support 11'. and may be held between the outer circumferential surface of the fitting portion 37 of the lid member 3.
【００２９】 本考案を適用して培養される細胞等としては、特に制限されるものではなく、 例えば動物細胞としてリンパ芽球、バーキットクンパ腫、急性リンパ芽球性白血 病細胞、骨髄腫などのミエローマ細胞、またはこれらによるハイブリドーマ細胞 、Ｂａｌｂ／Ｃマウスの脾臓細胞とＮＳ−１細胞とのハイブリドーマ、黒色種由 来のＢＯＷＥＳ細胞、ハイブリドーマの作製に用いる親株であるミエローマ、例 えばＳＰ２／Ｏ−Ａｇ−１４やＰ３／ＮＳ１−Ａｇ−４−１などの浮遊増殖性細 胞、ヒト子宮ガン細胞、ＨｅＬａ、チャイニーズ−ハムスター−肺細胞Ｖ−７９ 、ヒト胎児肺細胞ＭＲＣ−５、チンパンジー肝繊維芽細胞、ヒト包皮細胞、ニワ トリ胎児繊維芽細胞、初代サル腎細胞、マウス転移繊維芽細胞、脳下垂体腫瘍細 胞、アフリカミドリザル腎細胞Ｖｅｒｏ、副腎腫瘍細胞などの接着依存性細胞、 殺腫瘍細胞活性を示す一群のキラー細胞であるＬＡＫ細胞（Lymphokine Activat ed Killer Cell）、ＴＩＬ細胞（Tumor Infiltrating Lymphocyte ）、ＣＴＬ細 胞（Cytotoxic T Lymphocyte）などを挙げることができる。 なお、本考案の細胞培養用容器は、培養槽本体内を攪拌することがないので剪 断力に弱い細胞等であっても培養することができる。[0029] There are no particular restrictions on the cells etc. that can be cultured by applying the present invention. For example, animal cells such as lymphoblasts, Burkitt Kumpakoma, and acute lymphoblastic leukemia. Disease cells, myeloma cells such as myeloma, or hybridoma cells derived from these , a hybridoma of Balb/C mouse spleen cells and NS-1 cells, derived from melanoma BOWES cells, myeloma which is the parent line used for hybridoma production, e.g. For example, planktonic proliferative cells such as SP2/O-Ag-14 and P3/NS1-Ag-4-1. cells, human uterine cancer cells, HeLa, Chinese hamster lung cells V-79 , human fetal lung cells MRC-5, chimpanzee liver fibroblasts, human foreskin cells, chicken Fetal avian fibroblasts, primary monkey kidney cells, mouse metastatic fibroblasts, pituitary tumor cells adhesion-dependent cells such as cells, African green monkey kidney cells Vero, and adrenal gland tumor cells; LAK cells (Lymphokine Activat ed Killer Cell), TIL Cell (Tumor Infiltrating Lymphocyte), CTL Cell Examples include Cytotoxic T Lymphocytes. Note that the cell culture container of the present invention does not require any agitation within the culture tank, so it cannot be pruned. Even cells that are susceptible to shear force can be cultured.
【００３０】 ＜実験例＞実験例 培養細胞：抗組織プラスミノーゲンアクチベーター（抗ＴＰＡ）モノクローナ ル抗体産生ハイブリドーマ 培養液：酸素および炭酸ガスを溶存し、10容量％牛胎児血清を含むＲＰＭＩ１ ６４０ 粒子（攪拌子）：「バイオシロン」（ポリスチレン，粒径160 〜300 μｍ，日本 インターメッド社製） 400 mlの培養槽本体を有してなる上記実施例の細胞培養用容器に、乾燥重量で 0.6 ｇの粒子を入れて培養液を満たし、これに２×104 個／mlになるよう培養細 胞を播取し、空気が入らないよう密封した後、温度37℃の環境下、15日間に亘り 、回転数15r.p.m.で培養槽本体を図１に示すＸＹを軸として回転させて培養を行 った。 この間において牛胎児血清を含まない２リットルのＲＰＭＩ１６４０培養液を 循環させ、２日毎に新しい培養液ととり換えた。 酸素はシリコンチューブ（１×1.5 mm，４ｍ）よりなる酸素供給管に５容量％ 炭酸ガスと95容量％空気の混合ガスを通して供給した。 この結果、ハイブリドーマは順調に増殖し、７日目には２×107 個／mlに達し 、15日目までこの密度を維持した。<Experimental Example> Experimental Example Cultured cells: Anti-tissue plasminogen activator (anti-TPA) monoclonal antibody-producing hybridoma Culture medium: RPMI1 640 particles containing dissolved oxygen and carbon dioxide gas and 10% by volume fetal bovine serum ( Stirrer): "Biocilon" (polystyrene, particle size 160 to 300 μm, manufactured by Nihon Intermed Co., Ltd.) 0.6 g in dry weight was added to the cell culture container of the above example having a 400 ml culture tank body. Particles were added to fill the culture medium, and cultured cells were seeded in this at a concentration of 2 x 104 cells/ml. After sealing to prevent air from entering, the culture medium was incubated at a temperature of 37℃ for 15 days. Culture was carried out by rotating the culture tank main body around XY shown in FIG. 1 at a rotation speed of 15 rpm. During this period, 2 liters of RPMI 1640 culture solution containing no fetal bovine serum was circulated and replaced with fresh culture solution every 2 days. Oxygen was supplied through a mixed gas of 5% by volume carbon dioxide gas and 95% by volume air through an oxygen supply pipe made of a silicone tube (1×1.5 mm, 4 m). As a result, the hybridomas proliferated smoothly, reaching 2 x 10 7 cells/ml on the 7th day, and maintained this density until the 15th day.
【００３１】比較実験例 培養細胞：抗組織プラスミノーゲンアクチベーター（抗ＴＰＡ）モノクローナ ル抗体産生ハイブリドーマ 液体培養液：酸素および炭酸ガスを溶存し、10容量％牛胎児血清を含むＲＰＭ Ｉ１６４０ 市販のローラボトルに400 mlの培養液を入れ、これに２×104 個／mlになるよ う培養細胞を播取し、温度37℃の環境下、10日間に亘り、回転数５r.p.m.でロー ラボトルを回転させて培養を行った。５日目に半分の培養液の交換を行った。 この結果、ハイブリドーマは除々に増殖し、７日目には１×106 個／mlに達し たが、それ以上は増殖せず、ハイブリドーマは除々に死滅した。 Comparative Experimental Example Cultured cells: Anti-tissue plasminogen activator (anti-TPA) monoclonal antibody-producing hybridoma Liquid culture medium: RPM I1640 containing dissolved oxygen and carbon dioxide gas and 10% by volume fetal bovine serum Commercially available roller bottle Pour 400 ml of culture solution into the container, seed cultured cells at 2 x 104 cells/ml, and rotate the roller bottle at a rotation speed of 5 rpm for 10 days at a temperature of 37°C. The cells were then cultured. Half of the culture medium was replaced on the 5th day. As a result, the hybridomas gradually proliferated and reached 1×10 6 cells/ml on the 7th day, but did not proliferate any further and the hybridomas gradually died.
【００３２】[0032]
【考案の効果】 本考案によれば、培養槽本体を構成する槽支持体が、蓋部材の嵌合部と嵌合し て支持され、半透膜部材の一端が嵌合部および槽支持体の周側面によって挟持さ れているので、コンパクトな構成でありながら新しい培養液を培養を継続しなが ら培養槽本体内の細胞に連続的または間けつ的に供給することができ、高い密度 で細胞等を培養することができる。 また、半透膜部材の一端は液密に挟持されているので、培養槽本体内の細胞等 が培養槽本体外へ漏れることはない。 更に、槽支持体は、嵌合部に嵌合支持されているので、培養槽本体を蓋部材に 容易に着脱することができ、組立が容易である。[Effect of the idea] According to the present invention, the tank support that constitutes the culture tank body fits into the fitting portion of the lid member. and one end of the semipermeable membrane member is held between the fitting part and the circumferential side of the tank support. Because it has a compact structure, it is possible to continue culturing with new culture medium. can be supplied continuously or intermittently to the cells in the culture tank, resulting in high density Cells, etc. can be cultured. In addition, since one end of the semipermeable membrane member is sandwiched in a liquid-tight manner, cells inside the culture tank can be will not leak out of the culture tank body. Furthermore, since the tank support is fitted and supported by the fitting part, the culture tank body can be attached to the lid member. It can be easily attached and detached and is easy to assemble.
【図１】本考案の細胞培養用容器の一実施例を示す説明
用断面図である。FIG. 1 is an explanatory cross-sectional view showing one embodiment of the cell culture container of the present invention.
【図２】本考案の細胞培養用容器の他の実施例を示す説
明用断面図である。FIG. 2 is an explanatory sectional view showing another embodiment of the cell culture container of the present invention.
【図３】本考案の細胞培養用容器の他の実施例を示す説
明用断面図である。FIG. 3 is an explanatory sectional view showing another embodiment of the cell culture container of the present invention.
【図４】本考案の細胞培養用容器（円筒型）の好ましい
内径の比を示すグラフである。FIG. 4 is a graph showing a preferable inner diameter ratio of the cell culture container (cylindrical type) of the present invention.
１ 培養槽本体 ２ 外筒容器 ３ 蓋部材 ４ 酸素供給管 ５ リング部材 ５Ａ 螺合部 ６ Ｏリング 11 槽支持体 12 窓部 13 半透膜部材 15 底板部材 21 開口部 21Ａ 螺合部 23 Ｏリング溝 31 細胞供給口 32 酸素供給口 33 酸素排出口 32ａ 外部口金 32ｂ 内部口金 33ａ 外部口金 33ｂ 内部口金 34 培養液供給口 35 培養液排出口 34ａ 外部口金 35ａ 外部口金 36 内面 37 嵌合部 38 止め栓 41 端部 42 端部 1 Culture tank body 2 Outer cylinder container 3 Lid member 4 Oxygen supply pipe 5 Ring member 5A threaded part 6 O-ring 11 Tank support 12 Window section 13 Semipermeable membrane member 15 Bottom plate member 21 Opening 21A Threaded part 23 O-ring groove 31 Cell supply port 32 Oxygen supply port 33 Oxygen outlet 32a External cap 32b Internal cap 33a External cap 33b Internal cap 34 Culture medium supply port 35 Culture medium outlet 34a External cap 35a External cap 36 Inner side 37 Fitting part 38 Stopcock 41 End 42 End
Claims (1)
支持体および当該槽支持体の窓部を塞ぐよう設けられた
筒状の半透膜部材よりなる培養槽本体と、当該培養槽本
体が格納された、開口部を有する外筒容器と、当該外筒
容器の開口部を気密に塞ぐ蓋部材とにより構成され、前
記槽支持体は、蓋部材の内面に形成された嵌合部と嵌合
して支持され、前記半透膜部材は、その一端が前記嵌合
部および前記槽支持体の周側面によって液密に挟持され
ていることを特徴とする細胞培養用容器。Claim 1: A culture tank main body comprising a cylindrical tank support having a window formed on its peripheral side and a cylindrical semipermeable membrane member provided to close the window of the tank support; It is composed of an outer cylindrical container having an opening in which the culture tank body is stored, and a lid member that airtightly closes the opening of the outer cylindrical container, and the tank support is formed by a fitting formed on the inner surface of the lid member. A container for cell culture, characterized in that the semipermeable membrane member is supported by being fitted into the fitting part, and one end of the semipermeable membrane member is fluid-tightly sandwiched between the fitting part and the circumferential side of the tank support.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2219291U JPH04110500U (en) | 1991-03-14 | 1991-03-14 | Cell culture container |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2219291U JPH04110500U (en) | 1991-03-14 | 1991-03-14 | Cell culture container |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH04110500U true JPH04110500U (en) | 1992-09-25 |
Family
ID=31907736
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2219291U Withdrawn JPH04110500U (en) | 1991-03-14 | 1991-03-14 | Cell culture container |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH04110500U (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06233671A (en) * | 1993-02-08 | 1994-08-23 | Sumitomo Bakelite Co Ltd | Culture container |
| JP2004535826A (en) * | 2001-07-31 | 2004-12-02 | アーハードット，ウルスラ | Bioreactor with device whose wall is deformable |
| WO2019182093A1 (en) * | 2018-03-23 | 2019-09-26 | 住友ベークライト株式会社 | Culture vessel |
-
1991
- 1991-03-14 JP JP2219291U patent/JPH04110500U/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06233671A (en) * | 1993-02-08 | 1994-08-23 | Sumitomo Bakelite Co Ltd | Culture container |
| JP2004535826A (en) * | 2001-07-31 | 2004-12-02 | アーハードット，ウルスラ | Bioreactor with device whose wall is deformable |
| WO2019182093A1 (en) * | 2018-03-23 | 2019-09-26 | 住友ベークライト株式会社 | Culture vessel |
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