JPS63291580A - Culture of plant tissue and bioreactor using therefor - Google Patents
Culture of plant tissue and bioreactor using thereforInfo
- Publication number
- JPS63291580A JPS63291580A JP62123746A JP12374687A JPS63291580A JP S63291580 A JPS63291580 A JP S63291580A JP 62123746 A JP62123746 A JP 62123746A JP 12374687 A JP12374687 A JP 12374687A JP S63291580 A JPS63291580 A JP S63291580A
- Authority
- JP
- Japan
- Prior art keywords
- culture
- nonwoven cloth
- nonwoven fabric
- vessel
- cell
- 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.)
- Pending
Links
- 239000007788 liquid Substances 0.000 claims abstract description 22
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 13
- 238000009423 ventilation Methods 0.000 claims abstract description 4
- 239000004745 nonwoven fabric Substances 0.000 claims description 27
- 238000000034 method Methods 0.000 claims description 8
- 238000004161 plant tissue culture Methods 0.000 claims description 3
- 230000004069 differentiation Effects 0.000 abstract description 9
- 230000035755 proliferation Effects 0.000 abstract description 3
- 239000002689 soil Substances 0.000 abstract description 3
- 229920000297 Rayon Polymers 0.000 abstract description 2
- 239000011358 absorbing material Substances 0.000 abstract description 2
- 239000002964 rayon Substances 0.000 abstract description 2
- 239000004744 fabric Substances 0.000 abstract 8
- 238000007599 discharging Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 16
- 239000002609 medium Substances 0.000 description 15
- 241000196324 Embryophyta Species 0.000 description 13
- 230000012010 growth Effects 0.000 description 5
- 239000011148 porous material Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 4
- 229920000742 Cotton Polymers 0.000 description 3
- 239000004677 Nylon Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 229920001778 nylon Polymers 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 2
- 244000178937 Brassica oleracea var. capitata Species 0.000 description 2
- 244000000626 Daucus carota Species 0.000 description 2
- 235000002767 Daucus carota Nutrition 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 239000004743 Polypropylene Substances 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- -1 polypropylene Polymers 0.000 description 2
- 229920001155 polypropylene Polymers 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- OVSKIKFHRZPJSS-UHFFFAOYSA-N 2,4-D Chemical compound OC(=O)COC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-UHFFFAOYSA-N 0.000 description 1
- 229920002148 Gellan gum Polymers 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 1
- IDCBOTIENDVCBQ-UHFFFAOYSA-N TEPP Chemical compound CCOP(=O)(OCC)OP(=O)(OCC)OCC IDCBOTIENDVCBQ-UHFFFAOYSA-N 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000000679 carrageenan Substances 0.000 description 1
- 235000010418 carrageenan Nutrition 0.000 description 1
- 229920001525 carrageenan Polymers 0.000 description 1
- 229940113118 carrageenan Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000216 gellan gum Substances 0.000 description 1
- 235000010492 gellan gum Nutrition 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 229940064880 inositol 100 mg Drugs 0.000 description 1
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 1
- 229960001669 kinetin Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000002984 plastic foam Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000012780 transparent material Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は、一部が液体培地に浸漬した不織布を用いて植
物カルス細胞及び不定芽を増殖分化させる植物の組織培
養法及びこれに用いるバイオリアクターに関する。Detailed Description of the Invention [Field of Industrial Application] The present invention relates to a plant tissue culture method for growing and differentiating plant callus cells and adventitious buds using a nonwoven fabric partially immersed in a liquid medium, and a biotechnology method used therein. Regarding reactors.
従来、植物の組織培養法としては、培地に寒天等を用い
て固形化した固形培地による方法と液体培地による方法
とがある。BACKGROUND ART Conventionally, there are two methods for culturing plant tissue: a method using a solid medium made of agar or the like, and a method using a liquid medium.
固形培地では供給される栄養量が固定されるため植物ｍ
織の増殖分化にしたがって栄養の量、質共に変化させる
ことができない。液体培地は振とう或いは通気撹拌によ
り溶存酸素を供給しながら培養するが、この方法による
と固形培地による培養に比し、植物細胞が分散し培養細
胞中に産生物の蓄積が少なく、器官などの分化も抑制さ
れる傾向がある。また、連続的に培養できないという欠
点もある。In a solid medium, the amount of nutrients supplied is fixed, so the plant m
Both the quantity and quality of nutrients cannot be changed as the tissue proliferates and differentiates. The liquid medium is cultured while supplying dissolved oxygen by shaking or aerating the aeration, but with this method, compared to culture using a solid medium, the plant cells are dispersed, there is less accumulation of products in the cultured cells, and the growth of organs, etc. Differentiation also tends to be suppressed. Another drawback is that it cannot be cultured continuously.
そこで、液体培地の栄養の量及び質を変化させることが
できる長所と寒天培地の植物細胞が分散せず産生物が蓄
積され、分化が促進される長所とを併有し、連続運転で
きる植物組織の培養手段が求められていた。Therefore, a plant tissue that can be operated continuously has the advantage of being able to change the quantity and quality of nutrients in a liquid medium, and the advantage of an agar medium in that plant cells do not disperse, accumulating products and promoting differentiation. There was a need for a cultivation method.
本発明は上記問題を解決することを目的とし、その構成
は、液体培地に不織布を浸漬し、該不織布及び／又は該
不織布に付着して成長した植物細胞の一部を気体状空気
に接触させることによりカルス細胞又は不定芽を増殖分
化させることを特徴とし、更に、培養液供給口と培養液
排出口とを有する培養容器に、一部が液体培地に浸漬し
一部が空気中に露出するように不織布を敷き、培養液循
環装置と連結したことを特徴とする。The present invention aims to solve the above-mentioned problems, and has a structure in which a nonwoven fabric is immersed in a liquid medium, and the nonwoven fabric and/or a part of the plant cells that have grown attached to the nonwoven fabric are brought into contact with gaseous air. The method is characterized by causing callus cells or adventitious buds to proliferate and differentiate, and furthermore, in a culture container having a culture solution supply port and a culture solution outlet, a part is immersed in a liquid medium and a part is exposed to the air. It is characterized by being lined with non-woven fabric and connected to a culture solution circulation device.
本発明に係る不織布は、植物組織の増殖に悪影響を与え
ない吸水性の素材であればよく、レーヨン、ナイロンコ
ツトン、ポリプロピレン、ナイロン、ナイロン綿、レー
ヨン−テトロン、テトロン等が挙げられる。これら不織
布はそのまま使用してもよいが、縦横の格子状の切込み
を設けると不織布上に成長分化した植物組織を切込みに
従って分割し、そのまま土壌に移植することができる。The nonwoven fabric according to the present invention may be any water-absorbing material that does not adversely affect the growth of plant tissues, and examples thereof include rayon, nylon cotton, polypropylene, nylon, nylon cotton, rayon-tetron, and tetron. These nonwoven fabrics may be used as they are, but if vertical and horizontal lattice-like cuts are provided, plant tissue that has grown and differentiated on the nonwoven fabric can be divided according to the cuts and transplanted into the soil as is.
切込みは刃物による切断その他の物理的手段或いは薬品
等化学的手段により不織布に劣化部分を線状に設けるこ
とにより得られる。切込みは格子状に限らず一方向でも
よく、斜めに交叉する２方向以上の線を設けてもよい。The incision can be obtained by providing a linearly deteriorated portion in the nonwoven fabric by cutting with a knife or other physical means, or by chemical means such as chemicals. The cuts are not limited to a lattice shape, and may be made in one direction, or lines in two or more directions that intersect diagonally may be provided.
また、固定化担体を用いて不織布上に植物細胞を固定す
ると連続運転が可能になる。固定化担体としては、ｋ−
カラギナン、アルギン酸、ジェランガム等が使用される
。Furthermore, continuous operation is possible when plant cells are immobilized on a nonwoven fabric using an immobilization carrier. As the immobilization carrier, k-
Carrageenan, alginic acid, gellan gum, etc. are used.
培養液は公知の培養液を用いるが、植物組織の増殖に伴
い特定の成分が吸収され、その成分を補給する培養液や
分化を促進する培養液を植物組織の発育状態に合わせで
供給することが好ましい。A known culture solution is used as the culture solution, but specific components are absorbed as the plant tissue grows, and a culture solution that replenishes those components or a culture solution that promotes differentiation should be supplied according to the growth state of the plant tissue. is preferred.
このようにするには、バイオリアクターに培養液の供給
口と排出口を設はポンプを用いて循環させればよい。ま
た、ポンプは連続運転するがバイオリアクターが小型の
場合は間歇運転であってもよい。To do this, the bioreactor may be provided with an inlet and an outlet for the culture solution, and a pump may be used to circulate the culture solution. Further, the pump is operated continuously, but if the bioreactor is small, it may be operated intermittently.
培養容器はシャーレのような扁平形状のものが好ましい
。気体状の空気を供給するには容器上部に培養液に浸漬
しない充分な空間を保ち、充分な溶存酸素を有する培養
液を供給すればよい。或いは通気孔を設け、好ましくは
この通気孔に細菌遮断用のフィルターを設ける。フィル
ターはポリプロピレン、テフロン等のプラスチック発泡
体被膜、綿栓等細菌の混入を防止できるものであればよ
く孔径が多少大きくとも厚みがあれば遮断可能であり、
また素材が疎水性であれば孔径に比して細菌遮断効果が
大きい。一般には最大孔径０．２２μｍ程度が好ましい
。The culture container preferably has a flat shape like a petri dish. To supply gaseous air, it is sufficient to maintain a sufficient space above the container so that the container is not immersed in the culture solution, and to supply a culture solution with sufficient dissolved oxygen. Alternatively, a vent hole is provided, and preferably a filter for blocking bacteria is provided in the vent hole. The filter can be anything that can prevent bacteria from entering, such as a plastic foam coating such as polypropylene or Teflon, or a cotton plug, and even if the pore size is somewhat large, it can be blocked as long as it is thick.
Furthermore, if the material is hydrophobic, the bacteria-blocking effect is large compared to the pore size. Generally, the maximum pore diameter is preferably about 0.22 μm.
本発明は、液体培地に不織布を浸漬して植物細胞を不織
布内に定着させることにより、産生物が蓄積し難い液体
培地であっても産生物はカルス細胞や不定芽に効率よく
蓄積され、高密度培養が可能になる。不織布の上部が培
養液から露出していても、その吸水性により全体に培養
液が浸透しすると共に気体状の空気が常に供給されるた
め増殖が促進される。また、不織布上にカルス細胞を増
殖させたまま、培地組成を変更することが自由であるた
め、分化を促進させる培地組成を用いてそのまま分化さ
せることができる。充分に分化した後は不織布に切込み
を設けた場合は分離が容易であり、そのまま土壌に移植
できる長所がある。In the present invention, by soaking a nonwoven fabric in a liquid medium and fixing plant cells within the nonwoven fabric, even in a liquid medium in which it is difficult for products to accumulate, the products can be efficiently accumulated in callus cells and adventitious buds, resulting in a high growth rate. Density culture becomes possible. Even if the upper part of the non-woven fabric is exposed from the culture solution, its water absorbency allows the culture solution to permeate the entire surface, and gaseous air is constantly supplied, promoting growth. In addition, since the culture medium composition can be freely changed while callus cells are grown on the nonwoven fabric, differentiation can be directly carried out using a culture medium composition that promotes differentiation. After sufficient differentiation, separation is easy if cuts are made in the nonwoven fabric, which has the advantage of being able to be transplanted into soil as is.
本発明により液体培地を用いて植物細胞内に産生物を効
率よく蓄積させ、増殖分化を促進させることができる。According to the present invention, products can be efficiently accumulated in plant cells using a liquid medium, and proliferation and differentiation can be promoted.
〔実施例１〕及び〔比較例１〕
第１図は培養容器の断面図である。■は容器本体であり
、底部に培養液供給口２及び培養液排出口３を設けた。[Example 1] and [Comparative Example 1] FIG. 1 is a sectional view of a culture container. (2) is the container body, and a culture solution supply port 2 and a culture solution discharge port 3 were provided at the bottom.
４は透明素材からなる蓋であり、パツキン５を介して容
器本体１と気密に嵌合する。Reference numeral 4 denotes a lid made of a transparent material, which is airtightly fitted to the container body 1 via a gasket 5.
Ｍ４には通気孔６を穿設し最大孔径０．２２μｍのフイ
ルターフを設け、フィルター７を介して無菌空気を供給
した。８は不織布であり、培養液９に一部が浸漬してい
る。１０はカルス細胞であり、１）は不織布８に設けた
切込みである。M4 was provided with a ventilation hole 6 and a filter turf having a maximum pore diameter of 0.22 μm, and sterile air was supplied through the filter 7. 8 is a nonwoven fabric, and a part thereof is immersed in the culture solution 9. 10 is a callus cell, and 1) is a cut provided in the nonwoven fabric 8.
本実施例においては通気孔を蓋に設けたが、液面より上
であれば容器本体に設けてもよい。In this embodiment, the vent hole is provided in the lid, but it may be provided in the container body as long as it is above the liquid level.
第２図はフローシートである。１２は培養液密封容器で
あり、孔径０．２２μｍのフィルター１３を介して空気
供給管１４が開口している。更にポンプ１５を介して送
液管１６及びポンプ１７を介して受液管１８を設け、容
器本体１に設けた培養液供給口２及び培養液排出口３と
それぞれ接続した。Figure 2 is a flow sheet. Reference numeral 12 denotes a culture solution sealed container, into which an air supply pipe 14 is opened through a filter 13 having a pore size of 0.22 μm. Further, a liquid sending pipe 16 was provided via the pump 15, and a liquid receiving pipe 18 was provided via the pump 17, and these were connected to the culture solution supply port 2 and the culture solution discharge port 3 provided in the container body 1, respectively.
第１図に示す培養容器を用いて、ムラシゲ−スクーグ培
地（ＭｕｒａｓｈＬｇｅ ＆ Ｓｋｏｏｇ ）にイノシ
トール１００ｍｇ／ｌ、５ｕｃｒｏｓｅ ３０ ｇ／Ｉ
ｌｚ塩酸チアミン０ 、４ｍｇ ／ １．２．４ジクロ
ロフ工ノキシ酢酸１ｍｇ／ｌ、カイネチンｌＢ／Ｊを添
加した培地に、従来法により誘導したニンジンカルス細
胞を増殖させた。成長度を第１表に示す。Using the culture container shown in Figure 1, inositol 100 mg/l and 5ucrose 30 g/l were added to Murashige-Skoog medium (MurashLge & Skoog).
Carrot callus cells induced by a conventional method were grown in a medium supplemented with lzthiamine hydrochloride 0.4 mg/1.2.4 dichlorophenoxyacetic acid 1 mg/l and kinetin IB/J. The growth rate is shown in Table 1.
比較例１として、不織布を用いない以外は同一の条件で
同一のニンジンカルス細胞を培養した結果を第１表に併
記する。As Comparative Example 1, the same carrot callus cells were cultured under the same conditions except that a nonwoven fabric was not used. The results are also shown in Table 1.
第 １ 表
〔実施例２〕及び〔比較例２〕
常法により誘導したイネ及びレッドキャベツのカルス細
胞を用い実施例１と同様の条件で培養し、その結果を第
２表に記載した。また、比較例１と同様にしてイネ及び
レッドキャベツのカルス細胞を培養し、その結果を第２
表に併記した。Table 1 [Example 2] and [Comparative Example 2] Rice and red cabbage callus cells induced by conventional methods were cultured under the same conditions as in Example 1, and the results are shown in Table 2. In addition, callus cells of rice and red cabbage were cultured in the same manner as in Comparative Example 1, and the results were used in a second experiment.
Also listed in the table.
第 ２ 表Table 2
図面は本発明の実施例を示し、第１図は培養容器の断面
図、第２図はフローシートである。
図面中、符号
１は容器本体、２は培養液供給口、３は培養液排出口、
４は蓋、６は通気孔、７．１３はフィルター、８は不織
布、９は培養液、１０はカルス細胞、１）は切込み、１
２は培養液密封容器、１６は送液管、１８は受液管、１
５．１７はポンプである。
特許出願人 エヌオーケー株式会社
代理人弁理士 吉 １）俊 夫（外１
氾１図The drawings show examples of the present invention, with FIG. 1 being a cross-sectional view of a culture container and FIG. 2 being a flow sheet. In the drawing, 1 is the container body, 2 is the culture solution supply port, 3 is the culture solution outlet,
4 is a lid, 6 is a ventilation hole, 7.13 is a filter, 8 is a nonwoven fabric, 9 is a culture medium, 10 is a callus cell, 1) is a cut, 1
2 is a culture solution sealed container, 16 is a liquid sending tube, 18 is a liquid receiving tube, 1
5.17 is a pump. Patent applicant Yoshi, patent attorney, N.OK. 1) Toshio (external 1, flood 1)
Claims (6)
該不織布に付着して成長した植物細胞の一部を気体状空
気に接触させることによりカルス細胞又は不定芽を増殖
分化させる植物組織培養法。(1) Plant tissue culture in which callus cells or adventitious buds are proliferated and differentiated by immersing a nonwoven fabric in a liquid medium and exposing the nonwoven fabric and/or some of the plant cells that have grown attached to the nonwoven fabric to gaseous air. Law.
設けられている特許請求の範囲第１項記載の植物組織培
養法。(2) The plant tissue culture method according to claim 1, wherein the nonwoven fabric is provided with incisions that allow the nonwoven fabric to be easily divided.
に、一部が液体培地に浸漬し一部が空気中に露出するよ
うに不織布を敷き、培養液循環装置と連結したバイオリ
アクター。(3) A bioreactor in which a culture container with a culture solution supply port and a culture solution outlet is lined with a nonwoven fabric so that a portion is immersed in the liquid medium and a portion is exposed to the air, and connected to a culture solution circulation device. .
記載のバイオリアクター。(4) The bioreactor according to claim 3, wherein the culture container is provided with a ventilation hole.
求の範囲第４項記載のバイオリアクター。(5) The bioreactor according to claim 4, wherein the vent hole is provided with a cell-blocking filter.
設けた特許請求の範囲第３項ないし第５項のいずれかに
記載するバイオリアクター。(6) The bioreactor according to any one of claims 3 to 5, wherein the nonwoven fabric is provided with incisions that allow the nonwoven fabric to be easily divided.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62123746A JPS63291580A (en) | 1987-05-22 | 1987-05-22 | Culture of plant tissue and bioreactor using therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62123746A JPS63291580A (en) | 1987-05-22 | 1987-05-22 | Culture of plant tissue and bioreactor using therefor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63291580A true JPS63291580A (en) | 1988-11-29 |
Family
ID=14868299
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62123746A Pending JPS63291580A (en) | 1987-05-22 | 1987-05-22 | Culture of plant tissue and bioreactor using therefor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63291580A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0463584A (en) * | 1990-06-29 | 1992-02-28 | Fuji Photo Film Co Ltd | Bioreactor equipment |
| WO2003054138A1 (en) * | 2001-12-20 | 2003-07-03 | Applied Cell Biotechnologies, Inc. | Cell culture instrument and method of separating and subculturing cells |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57115181A (en) * | 1980-11-06 | 1982-07-17 | Bush Boake Allen Ltd | Preparation of plant metabolite |
| JPS61242522A (en) * | 1985-04-19 | 1986-10-28 | 株式会社ジャパンエナジー | Method for converting plant tissue to seedling |
| JPS62175170A (en) * | 1985-07-19 | 1987-07-31 | Nippon Steel Chem Co Ltd | Medium for tissue culture of plant |
-
1987
- 1987-05-22 JP JP62123746A patent/JPS63291580A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57115181A (en) * | 1980-11-06 | 1982-07-17 | Bush Boake Allen Ltd | Preparation of plant metabolite |
| JPS61242522A (en) * | 1985-04-19 | 1986-10-28 | 株式会社ジャパンエナジー | Method for converting plant tissue to seedling |
| JPS62175170A (en) * | 1985-07-19 | 1987-07-31 | Nippon Steel Chem Co Ltd | Medium for tissue culture of plant |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0463584A (en) * | 1990-06-29 | 1992-02-28 | Fuji Photo Film Co Ltd | Bioreactor equipment |
| WO2003054138A1 (en) * | 2001-12-20 | 2003-07-03 | Applied Cell Biotechnologies, Inc. | Cell culture instrument and method of separating and subculturing cells |
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