JP4529410B2 - Culture carrier, culture using the culture carrier, and method for producing the culture - Google Patents
Culture carrier, culture using the culture carrier, and method for producing the culture Download PDFInfo
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本発明は細胞層が多層の形で積層された積層細胞層、特に多種類の細胞層が多層の形で積層された積層細胞層を作製するために使用される培養担体、増殖用細胞が播種された前記の培養担体、増殖用細胞が播種された前記の培養担体の培養物とその培養方法、および前記培養物を使用した生体組織の再生方法に関する。 The present invention relates to a layered cell layer in which cell layers are laminated in a multilayered form, in particular, a culture carrier used for producing a laminated cell layer in which many types of cell layers are laminated in a multilayered form, and cells for proliferation are seeded. The present invention relates to the culture carrier described above, a culture of the culture carrier seeded with cells for proliferation, a culture method thereof, and a method of regenerating a living tissue using the culture.
従来、組織欠損部位への移植用材料として、代表的なものに人工皮膚が挙げられる。これら細胞膜の多くはコラーゲンに代表される生体由来の吸収性材料を細胞増殖の基材として細胞を三次元で培養することにより作製されている。しかしながら、このようにして作製した細胞膜は単層構造であり、増殖された細胞層が積層された形で存在する積層細胞層、特に多種類の細胞が増殖され、かつこれら細胞層が接着された積層細胞層を作製することは困難であった。 Conventionally, artificial skin is mentioned as a representative material for transplantation into a tissue defect site. Many of these cell membranes are produced by culturing cells in three dimensions using a living body-derived absorbent material typified by collagen as a base material for cell growth. However, the cell membrane produced in this way has a single-layer structure, and a laminated cell layer, in particular, a variety of cells grown in the form of a laminated cell layer, and these cell layers adhered. It was difficult to produce a laminated cell layer.
多種類の細胞が多層構造の積層状態で存在する積層細胞層を作製する方法としては、例えば単層の細胞層を積層する方法が用いられているが（非特許文献１）、この方法では単層の細胞層を連続的に積層する必要があり、多くの手間がかかり効率的でない。 As a method for producing a laminated cell layer in which many types of cells exist in a laminated state of a multilayer structure, for example, a method of laminating a single cell layer is used (Non-patent Document 1). It is necessary to continuously laminate cell layers, which is laborious and inefficient.
解決しようとする課題は、積層細胞層、特に多種類の細胞層が多層構造の積層状態で存在する積層細胞層を簡単に、かつ効率的に作製することができる培養担体、該培養担体を使用した培養物と培養方法、ならびに前記培養物を使用した生体組織の再生方法の提供することにある。なお、本発明でいう多種類の細胞層が多層構造の積層状態で存在する積層細胞層とは、複数の細胞層が積層状態で存在するものは全て含まれる。 The problem to be solved is a culture carrier capable of easily and efficiently producing a laminated cell layer, particularly a laminated cell layer in which many types of cell layers exist in a multilayered state, and uses the culture carrier. Another object of the present invention is to provide a culture and a culture method, and a method for regenerating a living tissue using the culture. In addition, the laminated cell layer in which a plurality of types of cell layers in the present invention exist in a laminated state includes all those in which a plurality of cell layers exist in a laminated state.
本発明の第１は、非多孔質層と多孔質層を有し、かつ生分解性材料で構成されたものであることを特徴とする細胞培養担体にある。 A first aspect of the present invention is a cell culture carrier characterized by having a non-porous layer and a porous layer and comprising a biodegradable material.
本発明の第２は、前記第１の細胞培養担体の複数個が多孔質層を上側として積み重ねて構成され、かつ最下層の非多孔質層の生分解速度が、他の非多孔質層や多孔質層の生分解速度に比較して小さいことを特徴とする細胞培養担体にある。 In the second aspect of the present invention, a plurality of the first cell culture carriers are stacked with the porous layer as the upper side, and the biodegradation rate of the lowermost non-porous layer is different from that of the other non-porous layer, The cell culture carrier is characterized by being smaller than the biodegradation rate of the porous layer.
本発明の第３は、前記第１または第２の培養担体の多孔質層中に増殖用細胞培養が播種されていることを特徴とする細胞培養担体にある。 According to a third aspect of the present invention, there is provided a cell culture carrier characterized in that a growth cell culture is seeded in the porous layer of the first or second culture carrier.
本発明の第４は、前記第３の細胞培養担体を培養した培養物にある。 A fourth aspect of the present invention resides in a culture obtained by culturing the third cell culture carrier.
本発明の第５は、前記第１あるいは第２の細胞培養担体の多孔質層中に増殖用細胞を播種後、該細胞培養担体を積み重ねた状態で増殖用細胞の培養を行うとともに最下位に存在する非多孔質層を除く非多孔質層と多孔質層の分解を行い、培養された増殖用細胞層を未分解の非多孔質層上に重力落下させることを特徴とする増殖用細胞の培養物の作製方法にある。 According to the fifth aspect of the present invention, after seeding the cells for proliferation in the porous layer of the first or second cell culture carrier, the cells for proliferation are cultured in a state where the cell culture carriers are stacked, and at the lowest level. A non-porous layer excluding an existing non-porous layer and a porous layer are decomposed, and the cultured cell layer for proliferation is dropped on the undegraded non-porous layer by gravity. It is in the production method of the culture.
本発明の第６は、前記第４の培養物を生体に移植して細胞増殖させることを特徴とする生体組織の再生方法にある。 According to a sixth aspect of the present invention, there is provided a method for regenerating a living tissue, wherein the fourth culture is transplanted into a living body to proliferate cells.
以下、本発明の実施態様を図面に基づいて具体的に説明する。
１．細胞培養担体の構成
本実施態様の図２で示される細胞培養担体は、図１に示される多孔質層１と非多孔質層２を有し、生分解性材料で構成された培養担体およびこの培養担体と同様に多孔質層３と非多孔質層４を有し、生分解性材料で構成されている細胞培養担体の２個をそれぞれ多孔質層を上側として積み重ねたものである。本実施態様の細胞培養担体は上述のように２個の細胞培養担体を積み重ねたものであるが、本発明の培養担体の積み重ね数は目的とする積層細胞層の使用分野あるいは用途に応じて適宜決定され、例えば再生皮膚用の増殖細胞層を目的とする場合には、表皮細胞を播種した第１の培養担体、該第１の培養担体の上に真皮細胞を播種した第２の培養担体が積み重ねられ、さらに該第２の培養担体の上に血管内皮前駆細胞皮細胞を播種した第３の培養担体が積み重ねられた３個の細胞培養担体で構成される。
Embodiments of the present invention will be specifically described below with reference to the drawings.
1. Structure of Cell Culture Carrier The cell culture carrier shown in FIG. 2 of the present embodiment has a porous layer 1 and a non-porous layer 2 shown in FIG. Like the culture carrier, it has a porous layer 3 and a non-porous layer 4, and two cell culture carriers made of a biodegradable material are stacked with the porous layer as the upper side. The cell culture carrier of this embodiment is obtained by stacking two cell culture carriers as described above. The number of the culture carrier of the present invention is appropriately determined depending on the intended use field or application of the laminated cell layer. For example, when a proliferation cell layer for regenerated skin is intended, a first culture carrier seeded with epidermal cells, and a second culture carrier seeded with dermal cells on the first culture carrier are: It is composed of three cell culture carriers that are stacked and further stacked with a third culture carrier in which vascular endothelial precursor cell skin cells are seeded on the second culture carrier .
前記図２に示す細胞培養担体の多孔質層１と多孔質層３および非多孔質層２と非多孔質層４の生分解速度は、使用する生分解性材料の選択および／または多孔質層の孔形状あるいは多孔度によってコントロールすることができる。すなわち、多孔質層は非多孔質層に比較して生分解速度が速く、また多孔度が大きい程、生分解速度が速く、また多孔質層の多孔形状として、例えば連続多孔層の方が独立多孔気泡層に比較して生分解速度が速く、かつ栄養が供給され易い。
図２に示す本実施態様の細胞培養担体は、上述のように使用する生分解性材料の選択および／または多孔質層の孔形状あるいは多孔度をコントロールすることによって、最下層の非多孔質層４の生分解速度は他の非多孔質層２や多孔質層１，３の生分解速度に比較してもっとも小さく構成され所定の培養が完了した未分解であるのに対して、多孔質層１，３は生分解速度がもっとも大きく構成され所定の培養が完了する迄に分解する分解速度のものであり、さらに非多孔質層２は生分解速度が前記最下層の非多孔質層４と多孔質層１，３の間の大きさで、該層の上下層として存在する増殖用細胞が播種されている多孔質層１，３が所定の培養が完了する迄は前記多孔質層１，３の増殖用細胞が混合しないように分離層としての機能を奏するが、所定の培養が完了した時点で分解するものである。
The biodegradation rates of the porous layer 1 and the porous layer 3 and the non-porous layer 2 and the non-porous layer 4 of the cell culture carrier shown in FIG. 2 depend on the selection of the biodegradable material to be used and / or the porous layer. It can be controlled by the pore shape or porosity. That is, the biodegradation rate of the porous layer is faster than that of the non-porous layer, and the greater the porosity, the faster the biodegradation rate. Also, as the porous shape of the porous layer, for example, the continuous porous layer is independent. Compared with the porous cell layer, the biodegradation rate is high and nutrition is easily supplied.
The cell culture carrier of the present embodiment shown in FIG. 2 has a lowermost non-porous layer by selecting the biodegradable material to be used and / or controlling the pore shape or porosity of the porous layer as described above. The biodegradation rate of 4 is the smallest compared to the biodegradation rates of the other non-porous layer 2 and porous layers 1 and 3 and is undegraded after completion of the predetermined culture, whereas the porous layer Nos. 1 and 3 have the highest biodegradation rate and are decomposed until the predetermined culture is completed. Further, the non-porous layer 2 has a biodegradation rate that is the same as that of the lowermost non-porous layer 4. The porous layers 1 and 3 having a size between the porous layers 1 and 3 and in which the cells for proliferation existing as the upper and lower layers of the layer are seeded are maintained until the predetermined culture is completed. 3 functions as a separation layer so that the cells for proliferation are not mixed. It is to decompose when given culture is completed.
前記図２に示す細胞培養担体は、前記のような構成を採用することにより、以下のような優れた効果を奏することができる。
すなわち、所定の培養が完了した時点においては非多孔質層２や多孔質層１，３は生分解により分解させ、多孔質層１，３において培養された培養物は最下位の非多孔質層４に載持させ、かつ該非多孔質層４を所定の培養が完了した時点においても該非多孔質層４上に載持された培養物を保持するに十分な強度を維持しているので、該培養物の取扱い、例えば生体へ移植する等に際しての取扱い、あるいは移植の作業が簡単となるだけでなく、通常、培養物を生体組織に移植して組織の再生誘導を行う場合、血液細胞を除いて欠損部位組織の再生誘導のために足場を必要とするが、前記最下位の非多孔質層４は、生体組織の再生誘導における足場の機能を奏することができるので、別途生体組織の再生誘導に際して前記のような足場の機能を奏する材料を使用する必要がなく、生体組織の再生誘導を簡単に行うことができるという効果を奏することができる。
なお、本実施態様の細胞培養担体においては、細胞培養担体の円柱形状のものを採用しているが、円柱形状のものに限らず、非多孔質層および多孔質層を形成できるものであれば任意の形状のもの、例えば４角形や膜形状のものが挙げられる。
The cell culture carrier shown in FIG. 2 can achieve the following excellent effects by adopting the configuration as described above.
That is, when the predetermined culture is completed, the non-porous layer 2 and the porous layers 1 and 3 are decomposed by biodegradation, and the culture cultured in the porous layers 1 and 3 is the lowest non-porous layer. 4 and the non-porous layer 4 maintains a sufficient strength to hold the culture supported on the non-porous layer 4 even when the predetermined culture is completed. In addition to simplifying the handling of the culture, for example, when transplanting into a living body, or the operation of transplanting, in general, when transplanting the culture into a living tissue and inducing tissue regeneration, blood cells are excluded. Although the scaffold is required for the regeneration induction of the defect site tissue, the lowermost non-porous layer 4 can function as a scaffold for the regeneration induction of the biological tissue. At the same time, the function of the scaffold as described above It is not necessary to use a material that can achieve the effect that it is possible to easily perform playback induction of biological tissue.
In addition, in the cell culture carrier of this embodiment, the cylindrical shape of the cell culture carrier is adopted. However, the cell culture carrier is not limited to the cylindrical shape, and may be any one that can form a non-porous layer and a porous layer. An arbitrary shape, for example, a quadrangular shape or a film shape can be mentioned.
本実施態様を含めて本発明で採用可能な生分解性材料としては、大きく分けて合成高分子材料、天然高分子材料および無機材料がある。
高分子材料としては、ポリグリコール酸、ポリ乳酸、ポリカプロラクトン、乳酸−グリコール酸共重合体、乳酪−カプロラクトン共重合体等が挙げられ、特に前記共重合体はその共重合成分の共重合比を変えることにより、その特性、例えば前記生分解性の速度を変更することができる。
天然高分子材料としては、コラーゲン、フィブロネクチン、ゼラチン、キチン、キトサン、ヒアルロン酸、アルギン酸等が挙げられる。また、これら天然高分子材料、コラーゲン、ゼラチン等は架橋処理することにより、その生分解性を任意に変更することができる。その他、生分解性無機材料としては、リン酸三カルシウム、炭酸カルシウムが挙げられる。
Biodegradable materials that can be employed in the present invention including this embodiment are broadly classified into synthetic polymer materials, natural polymer materials, and inorganic materials.
Examples of the polymer material include polyglycolic acid, polylactic acid, polycaprolactone, lactic acid-glycolic acid copolymer, and dairy dairy-caprolactone copolymer. In particular, the copolymer has a copolymerization ratio of its copolymer components. By changing the characteristics, for example the biodegradability rate can be changed.
Examples of natural polymer materials include collagen, fibronectin, gelatin, chitin, chitosan, hyaluronic acid, and alginic acid. Further, these natural polymer materials, collagen, gelatin and the like can be arbitrarily changed in their biodegradability by crosslinking treatment. In addition, examples of the biodegradable inorganic material include tricalcium phosphate and calcium carbonate.
２．細胞培養担体の作製法
本発明の培養担体の作製方法としては、例えば以下のようなものが挙げられる。
（１）多孔質体の一部を溶剤処理または熱処理して非多孔質化する方法
（２）非多孔質体の一部を凍結乾燥処理して多孔質化する方法
（３）多孔質層層と非多孔質層を接合する方法
前記（１）の方法で使用する多孔質体としては、例えば生分解性材料を発泡剤を使用して発泡させた発泡体が挙げられ、この発泡体の発泡条件あるいは使用する発泡剤の種類を選択することにより、適宜その多孔度および／または孔形状をコントロールすることができる。また、多孔質層における孔の大きさは、５０〜５００μｍが好ましい。多孔質層における孔の大きさが５０未満では、細胞・培養液が入りにくくなり、十分に細胞が育たない。５００μｍを超えると強度が弱くなる。
2. Production method of cell culture carrier Examples of the production method of the culture carrier of the present invention include the following.
(1) Method of making part of porous body non-porous by solvent treatment or heat treatment (2) Method of making part of non-porous body lyophilized to make porous (3) Porous layer layer The porous body used in the method (1) includes, for example, a foam obtained by foaming a biodegradable material using a foaming agent. Foaming of this foam By selecting the conditions or the type of blowing agent used, the porosity and / or pore shape can be appropriately controlled. The pore size in the porous layer is preferably 50 to 500 μm. When the size of the pores in the porous layer is less than 50, it becomes difficult for cells / culture medium to enter, and the cells do not grow sufficiently. If it exceeds 500 μm, the strength becomes weak.
３．前記細胞培養担体を利用した増殖用細胞の培養方法
目的の増殖細胞層の層数に応じた数の前記細胞培養担体を準備し、各細胞培養担体の多孔質層の孔中に目的の増殖細胞を得るために必要な増殖用細胞を播種した。この増殖用細胞の播種は、例えば前記増殖用細胞を滅菌、例えばγ線滅菌した前記細胞培養担体の多孔質層面上に供給した後、該供給された増殖用細胞を多孔質層中に埋入させることにより行うことができる。前記埋入手段としては、例えば前記増殖用細胞を播種した細胞培養担体を遠心させる遠心回転手段、あるいは滅菌された圧力ガスによる圧入手段が挙げられる。
また、前記増殖用細胞は前記埋入手段によっては多孔質層の表面部分にしか埋入できない場合があるとしても、生分解性材料の分解に伴い多孔質層内部にも埋入、あるいは増殖が可能となる。
3. Method of culturing cells for proliferation using the cell culture carrier The number of the cell culture carriers according to the number of layers of the target cell layer is prepared, and the target cells are proliferated in the pores of the porous layer of each cell culture carrier Cells necessary for growth were seeded. The seeding of the cells for proliferation is performed by, for example, supplying the cells for proliferation to the porous layer of the cell culture carrier sterilized, for example, sterilized by γ-ray, and then embedding the supplied cells for proliferation in the porous layer. Can be performed. Examples of the embedding means include a centrifugal rotating means for centrifuging the cell culture carrier seeded with the proliferation cells, or a press-fitting means using a sterilized pressure gas.
Further, even though the proliferation cells may be embedded only in the surface portion of the porous layer depending on the embedding means, they are also embedded in the porous layer or proliferated as the biodegradable material is decomposed. It becomes possible.
上述のようにして増殖用細胞を播種した細胞培養担体を必要な個数だけ積み重ね、該積み重ね状態で培養液（例えば血清を含有した培養液）を有する容器中に浸漬して増殖用細胞層を培養し、培養養開始後、最下層の非多孔質層を除いた非多孔質層非および多孔質層が分解消失させ、前記多孔質層中で増殖された培養細胞層を最下層の非多孔質層上に重力落下させ積層細胞層を形成させる。前記のようにして作成した培養物は、それを生体に移植して細胞増殖させることにより生体組織の再生を行うことができる。 A necessary number of cell culture carriers seeded with proliferation cells as described above are stacked, and the proliferation cell layer is cultured by immersing it in a container having a culture solution (for example, a culture solution containing serum). After the culture culture is started, the non-porous layer and the porous layer excluding the lowermost non-porous layer are decomposed and disappeared, and the cultured cell layer grown in the porous layer is removed from the lowermost non-porous layer. Gravity falls on the layer to form a laminated cell layer. The culture prepared as described above can regenerate a living tissue by transplanting it into a living body and growing cells.
細胞培養担体および該培養担体を使用した細胞膜の作製方法 Cell culture carrier and method for producing cell membrane using the culture carrier
細胞培養担体１および該細胞培養担体を使用した積層細胞層の作製方法
図１および２に基づいて説明する。
分子量１０万の乳酸−カプロラクトン共重合体を１,４−ジオキサンに溶解し５％の溶液とし、この溶液を膜状で−５０℃にて凍結した。この膜状凍結物の片面のみを２５℃に２分置き溶解させた後に、凍結乾燥法を用いて片面のみが多孔質化した３個の乳酸−カプロラクトン共重合体膜（厚さ５００μｍ）を細胞培養担体として得た。得られた前記乳酸−カプロラクトン共重合体膜の少なくとも多孔質面は放電により親水化処理を行って細胞接着性を向上させた。さらにこの細胞培養担体の少なくとも多孔質層面、好ましくは全体をγ線滅菌した。上述のように親水化処理およびγ線滅菌を行った３個の細胞培養担体の多孔質面上に、それぞれ表皮細胞、真皮細胞、血管内皮前駆細胞をそれぞれ１．０×１０４個ずつ播種し、播種後、３００ｒｐｍで遠心することで多孔質内部細胞を埋入した。前記増殖用細胞を播種した膜状の細胞培養担体を積層して、該積層状態で１０％血清含有培養液中に浸漬して培養を行った。１５日後には多孔質部位はほぼ分解し、消失部分に細胞が増殖されていることが確認された。培養開始から２０日後、非多孔質層が分解され始め、各細胞膜の接着が見られた。
Cell culture carrier 1 and method for producing laminated cell layer using the cell culture carrier A description will be given based on FIGS.
A lactic acid-caprolactone copolymer having a molecular weight of 100,000 was dissolved in 1,4-dioxane to obtain a 5% solution, and this solution was frozen in a film at -50 ° C. After dissolving only one side of this membrane-like frozen material at 25 ° C. for 2 minutes, three lactic acid-caprolactone copolymer membranes (thickness 500 μm) having only one side made porous using freeze-drying were treated with cells. Obtained as a culture carrier. At least the porous surface of the obtained lactic acid-caprolactone copolymer film was subjected to a hydrophilic treatment by discharge to improve cell adhesion. Furthermore, at least the porous layer surface of this cell culture carrier, preferably the whole was sterilized by γ-rays. On the porous surfaces of the three cell culture carriers that have been subjected to hydrophilization treatment and γ-ray sterilization as described above, 1.0 × 10 4 each of epidermis cells, dermal cells, and vascular endothelial progenitor cells are seeded. After seeding, the porous inner cells were embedded by centrifugation at 300 rpm. The membrane-shaped cell culture carrier seeded with the proliferation cells was laminated and cultured in the laminated state by being immersed in a 10% serum-containing culture solution. After 15 days, it was confirmed that the porous part was almost decomposed and cells were grown in the disappeared part. After 20 days from the start of the culture, the non-porous layer began to be decomposed, and adhesion of each cell membrane was observed.
細胞培養担体２および該細胞培養担体を使用した積層細胞層の作製方法
前記実施例１において、前記表皮細胞および真皮細胞を播種する細胞培養担体は、分子量１０万の乳酸−カプロラクトン共重合体をポリグリコール酸に替えた以外は同様にして作製し、また、血管内皮前駆細胞を播種する細胞培養担体は実施例１と同様にして作製した。これら各細胞培養担体を実施例１と同様に積み重ねて培養した。本実施例の細胞培養担体は、ポリグリコール酸で構成された多孔質部位がほぼ分解し、消失部分に細胞が増殖されていることが確認された時点、さらにはポリグリコール酸で構成された非多孔質層が分解され始め、各細胞膜の接着が見られた時点においても、前記乳酸−カプロラクトン共重合体で構成された最下位の非多孔質は未分解であった。
なお、前記表皮細胞、真皮細胞および血管内皮前駆細胞の培養に適した培養液としては、それぞれ従来用いられているものを使用することができる。
Cell culture carrier 2 and method for producing a laminated cell layer using the cell culture carrier In Example 1, the cell culture carrier for seeding the epidermal cells and dermal cells is a polylactic acid-caprolactone copolymer having a molecular weight of 100,000. A cell culture carrier for seeding vascular endothelial progenitor cells was prepared in the same manner as in Example 1 except that glycolic acid was used. These cell culture carriers were stacked and cultured in the same manner as in Example 1. In the cell culture carrier of this example, when it was confirmed that the porous site composed of polyglycolic acid was almost decomposed and cells were grown in the disappeared part, and further non-constituted composed of polyglycolic acid. Even when the porous layer began to be decomposed and adhesion of each cell membrane was observed, the lowest non-porous material composed of the lactic acid-caprolactone copolymer was undecomposed.
In addition, as a culture solution suitable for culturing the epidermal cells, dermal cells, and vascular endothelial precursor cells, those conventionally used can be used.
本発明の培養担体を利用して、例えば皮膚再生用の積層細胞層層を製造することができる。すなわち、皮膚は角化細胞、色素細胞やメルケル細胞等で構成される表皮層、線維芽細胞と呼ばれる細胞でつくられた線維と基質で構成される真皮層および真皮の下にあり、脂肪細胞を含み、血管や神経幹も走っている下皮を基本構成とするものであるが、皮膚細胞は他の生体組織、またこれに代わる物質に付着して増殖し，平面上に層を作る性質があり、従来、細胞培養床としてポリスチレンや合成高分子物質に天然高分子物質をコーティングしたもの等を利用して細胞増殖が行われている。また、本発明の培養担体を利用すると、皮膚再生用の積層細胞層の他、血管再生用の積層細胞層、内臓臓器再生用の積層細胞層等を容易に作成することができる。 For example, a laminated cell layer layer for skin regeneration can be produced using the culture carrier of the present invention. That is, the skin is in the epidermis layer composed of keratinocytes, pigment cells, Merkel cells, etc., the dermis layer composed of fibers and substrates made of cells called fibroblasts and the dermis, and adipocytes Including the lower skin where blood vessels and nerve trunks are running, the skin cells adhere to other biological tissues and alternative substances and proliferate, creating a layer on a flat surface. Conventionally, cell growth is performed using a cell culture bed made of polystyrene or a synthetic polymer material coated with a natural polymer material. In addition, by using the culture carrier of the present invention, a laminated cell layer for regenerating blood vessels, a laminated cell layer for regenerating blood vessels, a laminated cell layer for regenerating visceral organs, and the like can be easily prepared.
１ 多孔質層
２ 非多孔質層
３ 多孔質層
４ 非多孔質層
DESCRIPTION OF SYMBOLS 1 Porous layer 2 Non-porous layer 3 Porous layer 4 Non-porous layer
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| US4787900A (en) * | 1982-04-19 | 1988-11-29 | Massachusetts Institute Of Technology | Process for forming multilayer bioreplaceable blood vessel prosthesis |
| JPH07236688A (en) * | 1994-02-28 | 1995-09-12 | Terumo Corp | Composite material of bioabsorptive plastic and collagen |
| JP2000507847A (en) * | 1996-03-04 | 2000-06-27 | キルシユ，アクセル | Coating film, molded article produced therefrom and method for producing the same |
| JP2000237298A (en) * | 1999-02-10 | 2000-09-05 | Isotis Bv | Cartilaginous tissue engineering |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4787900A (en) * | 1982-04-19 | 1988-11-29 | Massachusetts Institute Of Technology | Process for forming multilayer bioreplaceable blood vessel prosthesis |
| JPH07236688A (en) * | 1994-02-28 | 1995-09-12 | Terumo Corp | Composite material of bioabsorptive plastic and collagen |
| JP2000507847A (en) * | 1996-03-04 | 2000-06-27 | キルシユ，アクセル | Coating film, molded article produced therefrom and method for producing the same |
| JP2000237298A (en) * | 1999-02-10 | 2000-09-05 | Isotis Bv | Cartilaginous tissue engineering |
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