JP2018019654A - Culture product, and culture vessel - Google Patents
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Abstract
Description
本発明は細胞培養に係り、特に接着細胞などを簡単に培養する技術に関する。 The present invention relates to cell culture, and more particularly to a technique for easily culturing adherent cells and the like.
細胞を生体外で培養する際には、その細胞種特異的な機能や活性の維持に適した培養環境が重要であり、その環境因子として、培養器材、器材表面のコート剤、培地、液性因子などがある。生体内において細胞が分化・増殖する場合には、細胞外マトリックスが足場として機能して組織の構築が行われるため、足場材となる培養用器材や、器材表面の細胞外マトリックス由来基質などを含むコート剤は、生体外での細胞培養において重要な役割を果たす。 When culturing cells in vitro, a culture environment suitable for maintaining the function and activity specific to the cell type is important. The environmental factors include culture equipment, coating agents on the equipment surface, medium, liquidity There are factors. When cells are differentiated and proliferated in vivo, the extracellular matrix functions as a scaffold to construct a tissue, and therefore includes a culture device as a scaffold, an extracellular matrix-derived substrate on the surface of the device, etc. The coating agent plays an important role in cell culture in vitro.
人工多能性幹細胞（以下、iPS細胞）は様々な細胞に分化できる多能性と無限に増殖できる自己複製能を有する細胞（非特許文献１）であるため、再生医療や創薬の分野においてiPS細胞の利用が期待されている。このiPS細胞を未分化能を維持したまま増殖させるためには、培養器材表面のコート剤や培地の組み合わせにより、適切な培養環境を維持することが特に重要となる（非特許文献２）。培養環境が適切でない場合には、iPS細胞の未分化の状態を維持できなかったり、細胞が死滅してしまったりすることがある。 Artificial pluripotent stem cells (hereinafter iPS cells) are pluripotent cells that can differentiate into various cells and self-replicating cells that can proliferate indefinitely (Non-patent Document 1). Use of iPS cells is expected. In order to proliferate the iPS cells while maintaining the undifferentiated ability, it is particularly important to maintain an appropriate culture environment by combining a coating agent on the surface of the culture equipment and a medium (Non-patent Document 2). If the culture environment is not appropriate, the undifferentiated state of iPS cells may not be maintained, or the cells may die.
未分化状態を維持する培養方法には、支持細胞であるフィーダ細胞と共培養するオンフィーダ培養法と、フィーダ細胞に替わる添加剤のみで未分化状態を維持するフィーダフリー培養法があり、支持細胞や細胞外マトリックス由来基質が必要である。更に、再生医療にiPS細胞を用いる際には、支持細胞由来の汚染や支持細胞自体の混入を防ぐ必要があるため、細胞外マトリックス由来基質の利用が不可欠である。このような技術に関連する特許文献としては、特許文献１がある。 Culture methods that maintain an undifferentiated state include an on-feeder culture method that co-cultures with feeder cells that are feeder cells, and a feeder-free culture method that maintains an undifferentiated state only with an additive that replaces feeder cells. And an extracellular matrix-derived substrate is required. Furthermore, when iPS cells are used in regenerative medicine, it is necessary to prevent contamination from feeder cells and contamination of the feeder cells themselves, and therefore it is essential to use an extracellular matrix-derived substrate. Patent Literature 1 is related to such a technique.
上述した細胞外マトリックス由来基質であるラミニンは、iPS細胞を未分化な状態で安定的に増殖培養するための基質として広く利用されているが、コート時に表面で安定化させるために時間を要する。また表面が乾燥するとその培養機能が低下する。また、コートした表面の乾燥保存が難しいため、使用直前のコーティングが必要であるという課題がある。 Laminin, which is an extracellular matrix-derived substrate described above, is widely used as a substrate for stably growing and culturing iPS cells in an undifferentiated state. However, it takes time to stabilize the surface at the time of coating. In addition, when the surface is dried, the culture function decreases. In addition, since it is difficult to dry and store the coated surface, there is a problem that a coating immediately before use is necessary.
本発明の目的は、上記の課題を解決し、作業工程を簡略化し、作業に費やす時間を低減することが可能な培養用生成物、及び培養容器を提供することにある。 The objective of this invention is providing the product for culture | cultivation which can solve said subject, simplifies a work process, and can reduce the time spent on work, and a culture container.
本発明においては、上記の課題を解決するため、細胞の培養用表面を有する培養用生成物であって、培養用表面は、細胞外マトリックス由来基質と、マルトース或いはラクトースの少なくとも1つ含む糖類を含む安定化剤とを有する培養用生成物を提供する。 In the present invention, in order to solve the above problems, a culture product having a cell culture surface, the culture surface comprising an extracellular matrix-derived substrate and a saccharide containing at least one of maltose or lactose. A culture product having a stabilizing agent is provided.
また、本発明においては、上記の課題を解決するため、培養容器であって、容器本体と、容器本体の表面に塗られた、細胞の培養用生成物とを備え、培養用生成物は、細胞外マトリックス由来基質と、マルトース或いはラクトースの少なくとも1つ含む糖類を含む安定化剤とを有する培養容器を提供する。 Further, in the present invention, in order to solve the above-mentioned problems, the culture vessel comprises a vessel body and a cell culture product applied to the surface of the vessel body, and the culture product comprises: A culture vessel having an extracellular matrix-derived substrate and a stabilizer containing a saccharide containing at least one of maltose or lactose is provided.
本発明により、細胞外マトリックス由来基質保持表面の乾燥保存が可能であり、培養使用直前の細胞外マトリックス由来基質のコーティング作業を省略可能であり、細胞培養開始時の時間と作業工程を短縮可能となる。 According to the present invention, the extracellular matrix-derived substrate holding surface can be dried and stored, and the coating operation of the extracellular matrix-derived substrate immediately before the use of culture can be omitted, and the time and work process at the start of cell culture can be shortened. Become.
以下、本発明の培養生成物、及び培養容器の実施形態について詳述する。本発明者が検討を重ねた結果、細胞外マトリックス由来基質を乾燥状態で保存した表面でも細胞培養が可能であり、簡便に使用できることを見出し、本発明の培養生成物を完成させるに至った。本発明は、iPS細胞で代表される接着細胞の機能を維持したまま培養できる表面を提供するものであり、細胞外マトリックス由来基質と安定化剤を表面に有する培養用生成物、及び培養容器を構成する。すなわち、細胞の培養用表面を有する培養用生成物であって、培養用表面は、細胞外マトリックス由来基質と、マルトース或いはラクトースの少なくとも1つ含む糖類を含む安定化剤を有する培養用生成物、及びそれを利用した培養容器を構成する。 Hereinafter, embodiments of the culture product and the culture container of the present invention will be described in detail. As a result of repeated studies by the present inventors, it has been found that cell culture is possible even on a surface where an extracellular matrix-derived substrate is stored in a dry state and can be used easily, and the culture product of the present invention has been completed. The present invention provides a surface that can be cultured while maintaining the function of adherent cells typified by iPS cells, and comprises a culture product having an extracellular matrix-derived substrate and a stabilizer on the surface, and a culture vessel. Configure. That is, a culture product having a cell culture surface, the culture surface having an extracellular matrix-derived substrate and a stabilizer containing a saccharide containing at least one of maltose or lactose, And a culture vessel using the same.
本発明は主として接着細胞を対象とし、接着細胞にはiPS細胞・ES細胞を含む幹細胞や体性幹細胞、各種初代培養細胞、株化細胞が含まれるが、この限りではない。また本発明は、細胞外マトリックス由来基質と安定化剤を含む培養用生成物であれば良く、細胞外マトリックス由来基質としては、ラミニン、コラーゲン、プロテオグリカン、エンタクチン/ニドゲン、フィブロネクチンなどの糖タンパク質や成長因子またはいずれかの混合物が挙げられるがこれに限定されない。細胞外マトリックス由来基質がラミニンの場合、ラミニンの全長を有しているもの、更には一部機能部位の断片などが含まれる。また本発明では特に、糖類を安定化剤として使用する。安定化剤として用いる糖類は、単糖、二糖、多糖類を、単独でも混合されて使用しても良く、特にマルトース、ラクトースを少なくとも1つ含むことが望ましい。 The present invention is mainly directed to adherent cells. Adherent cells include stem cells and somatic stem cells including iPS cells and ES cells, various primary cultured cells, and established cells, but are not limited thereto. The present invention may be any culture product containing an extracellular matrix-derived substrate and a stabilizer, and examples of the extracellular matrix-derived substrate include laminin, collagen, proteoglycan, entactin / nidogen, fibronectin and other glycoproteins and growths. Factors or mixtures of any include, but are not limited to. In the case where the extracellular matrix-derived substrate is laminin, those having the full length of laminin, and further partially fragmented functional sites are included. In the present invention, saccharides are particularly used as a stabilizer. The saccharides used as the stabilizer may be monosaccharides, disaccharides or polysaccharides, which may be used alone or in combination, and particularly preferably contain at least one of maltose and lactose.
本発明によって提供される培養用生成物が使用される培養容器の容器本体は、培養皿、マルチウェルプレート、培養フラスコ、および同様の培養用の容器が全て含まれる。この培養容器の容器本体は、培養用生成物に含まれる細胞外マトリックス由来基質の吸着や結合を可能にできる物質でできたものであり、ポリスチレン、ポリカーボネート、ポリプロピレン、ポリエチレン、ガラス、セルロース、フィブロインなど高分子材料が含まれ、これらの組み合わせを含むことが出来る。 The container body of the culture container in which the product for culture provided by the present invention is used includes a culture dish, a multiwell plate, a culture flask, and similar culture containers. The container body of this culture container is made of a substance that enables adsorption and binding of the extracellular matrix-derived substrate contained in the culture product, such as polystyrene, polycarbonate, polypropylene, polyethylene, glass, cellulose, fibroin, etc. Polymeric materials are included and can include combinations thereof.
本発明により提供される培養用生成物は、細胞外マトリックス由来基質をコーティング後に安定化剤を塗布する場合や、細胞外マトリックス由来基質と安定化剤を同時に塗布することによって作製される生成物を含む。 The culture product provided by the present invention is a product produced by applying a stabilizer after coating an extracellular matrix-derived substrate, or a product produced by simultaneously applying an extracellular matrix-derived substrate and a stabilizer. Including.
また、本発明の培養用生成物を生成するため、細胞外マトリックス由来基質や安定化剤を調製する溶液は、リン酸緩衝生理食塩水（PBS）やトリス緩衝生理食塩水など細胞培養に使用する緩衝液であれば、特にその種類は問わない。この安定化剤を含む溶液は器材表面にコーティング前に、好適には0.22μｍのフィルターでろ過滅菌を行う。 Moreover, in order to produce the culture product of the present invention, a solution for preparing an extracellular matrix-derived substrate or a stabilizer is used for cell culture such as phosphate buffered saline (PBS) or Tris buffered saline. If it is a buffer solution, the kind in particular will not ask | require. The solution containing the stabilizer is preferably sterilized by filtration with a 0.22 μm filter before coating on the surface of the equipment.
本発明は、安定化剤いずれかを0.008〜0.3 mg / cm2有する生成物であり、より好ましくは0.01〜0.2 mg / cm2有する生成物であり、さらに好ましくは0.03〜0.1 mg / cm2有する生成物である。これら0.03 mg、0.1 mg、0.2 mg / cm2は本発明によって提供される培養用生成物の安定化剤保持量の計測値であり、0.03 mg / cm2を示す生成物の安定化剤濃度は解析に用いた装置の定量限界である。ここで言う安定化剤保持量とは安定化剤を培養表面に添加し、一定時間静置後、上清を除去後に培養表面に残っている安定化剤の量を所定の方法で計測した量である。ここで言う所定の方法とは作製した培養用生成物上に超純水を添加し、24時間静置することで安定化剤を抽出し、その安定化剤抽出溶液を回収し、HPLC溶離液で希釈し、0.45μｍメンブレンフィルターでろ過し、測定試料として用いて測定する方法である。表面保持量はそれぞれの安定化剤を用いて作成した検量線によって導かれ、各表面から抽出された糖量を生成物の表面積単位で示した。0.01 mg、0.008 mgは検量線を用いて本発明作製時の安定化剤の濃度から概算される値である。 The present invention is a one stabilizer products with 0.008 to 0.3 mg / cm 2, more preferably products with 0.01 to 0.2 mg / cm 2, even more preferably 0.03 to 0.1 mg / cm 2 Product. These 0.03 mg, 0.1 mg, and 0.2 mg / cm 2 are measured values of the stabilizer retention amount of the culture product provided by the present invention, and the stabilizer concentration of the product showing 0.03 mg / cm 2 is This is the limit of quantification of the device used for the analysis. The amount of stabilizer retained here refers to the amount of stabilizer added to the culture surface, measured for the amount of stabilizer remaining on the culture surface after removing the supernatant after standing for a certain period of time. It is. The specified method here refers to adding the ultrapure water to the prepared culture product, and allowing it to stand for 24 hours to extract the stabilizer, collect the stabilizer extract solution, and recover the HPLC eluent. In this method, the sample is diluted with 1 and filtered through a 0.45 μm membrane filter and used as a measurement sample. The amount of surface retention was derived from a calibration curve prepared using each stabilizer, and the amount of sugar extracted from each surface was shown in terms of the surface area of the product. 0.01 mg and 0.008 mg are values estimated from the concentration of the stabilizer at the time of preparation of the present invention using a calibration curve.
＜細胞外マトリックス由来基質保持量の算出方法＞
安定化剤と同様に細胞外マトリックス由来基質の保持量は次のように測定した。作製した培養用生成物上に超純水を添加し、２４時間静置することで細胞外マトリックス由来基質を抽出した。細胞外マトリックス由来基質抽出溶液を回収し、HPLC溶離液で希釈し、0.45μｍメンブレンフィルターでろ過し、測定試料とした。
<Calculation method of extracellular matrix-derived substrate retention amount>
Similar to the stabilizer, the amount of extracellular matrix-derived substrate was measured as follows. Extrapure water was added onto the prepared culture product and left for 24 hours to extract an extracellular matrix-derived substrate. The extracellular matrix-derived substrate extraction solution was recovered, diluted with HPLC eluent, and filtered through a 0.45 μm membrane filter to obtain a measurement sample.
本発明によって提供される培養用生成物は、37℃以下で保存可能であり、より好ましくは25℃以下で保存可能であり、さらに好ましくは4℃以下で保存可能である。特に単糖の一種であるマンノースを安定化剤として用いた本発明によって提供される生成物は、4℃で3カ月間保存後もラミニンの機能を維持しており、使用直前にラミニンコーティングした表面と同等の培養が可能であった。また、ラクトースを安定化剤として用いた本発明によって提供される生成物は、4℃で1カ月間保存後もラミニンの機能を維持しており、さらには37℃で高湿度環境下で1カ月間保存後もラミニンの機能を維持しており、使用直前にラミニンコーティングした表面と同等の培養が可能であった。 The culture product provided by the present invention can be stored at 37 ° C. or lower, more preferably 25 ° C. or lower, and even more preferably 4 ° C. or lower. In particular, the product provided by the present invention using mannose, a monosaccharide as a stabilizer, maintains the function of laminin even after storage at 4 ° C. for 3 months, and the surface coated with laminin immediately before use. Cultivation equivalent to was possible. The product provided by the present invention using lactose as a stabilizer maintains the function of laminin even after storage at 4 ° C for 1 month, and further at 37 ° C for 1 month in a high humidity environment. The function of laminin was maintained even after storage for a long time, and it was possible to culture the same as the laminin-coated surface just before use.
なお、本発明において培養に使用する培養液は、使用する細胞について良好な成育が得られる一般的な細胞培養液であれば、特にその種類を問わない。 In addition, the culture solution used for culture in the present invention is not particularly limited as long as it is a general cell culture solution that can achieve good growth of the cells used.
以下、実施例により、本発明をより詳細に説明するが、本発明の技術範囲は以下の実施例に限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, the technical scope of this invention is not limited to a following example.
実施例１の培養用生成物の生成について順次説明する。安定化剤溶液は各種糖を15 mM〜300 mMの濃度になるようにPBS溶液で溶解した。完全に糖が溶けたことを確認後、0.22 μmのフィルターに通しろ過滅菌を行った。 The production of the culture product of Example 1 will be described sequentially. As the stabilizer solution, various sugars were dissolved in a PBS solution so as to have a concentration of 15 mM to 300 mM. After confirming that the sugar was completely dissolved, the solution was sterilized by filtration through a 0.22 μm filter.
細胞外マトリックス由来基質としてiMatrix-511（カタログ番号892011；ニッピ）を使用し、PBSを400 μL添加した培養器材表面（24 well plate）にiMatrix-511を2 μL添加し、4℃で終夜コーティングした。iMatrix-511のコーティング液を除去し、調製した安定化剤を含むPBS溶液を400 μL添加し、コーティングした後、除去し乾燥保存した器材を本実施例によって提供される生成物とした。また、調整した安定化剤を含むPBS溶液を400 μL培養器材表面に添加し、iMatrix-511を2 μL添加し4℃で終夜コーティングした。その安定化剤とiMatrix-511の混合液を除去し乾燥保存した器材を本実施例によって提供される生成物とした。比較例１として、PBSを400 μL添加した培養器材表面にiMatrix-511を2 μL添加し、4℃で終夜コーティングし、コーティング液を除去し乾燥保存した器材を作製した。比較例２として、PBSを400 μL添加した培養器材表面にiMatrix-511を2 μL添加し、4℃で終夜コーティングし、コーティング液を除去した器材を作製した。 Using iMatrix-511 (catalog number 892011; Nippi) as the extracellular matrix-derived substrate, add 2 μL of iMatrix-511 to the surface of the culture equipment (24 well plate) to which 400 μL of PBS was added, and coat at 4 ° C overnight. . The coating solution of iMatrix-511 was removed, and 400 μL of the PBS solution containing the prepared stabilizer was added, and after coating, the device removed and stored dry was used as the product provided by this example. Further, 400 μL of the PBS solution containing the prepared stabilizer was added to the surface of the culture equipment, and 2 μL of iMatrix-511 was added, followed by coating at 4 ° C. overnight. The product provided by this example was the equipment that had been removed from the mixture of the stabilizer and iMatrix-511 and stored dry. As Comparative Example 1, 2 μL of iMatrix-511 was added to the surface of the culture equipment to which 400 μL of PBS was added, and the coating was removed at 4 ° C. overnight. As Comparative Example 2, 2 μL of iMatrix-511 was added to the surface of the culture equipment to which 400 μL of PBS was added, and the equipment was coated overnight at 4 ° C. to remove the coating solution.
作製した器材上に超純水を添加し、２４時間静置することで安定化剤を抽出した。安定化剤抽出溶液を回収し、HPLC溶離液（アセトニトリル：超純水＝70：30 v/v）で希釈し測定試料とし、安定化剤の保持量を測定した。安定化剤の各濃度に応じた糖の保持量が検出された。これらの結果である器材表面の糖保持量を以下の表１に示す。 Ultrapure water was added onto the prepared equipment and left for 24 hours to extract the stabilizer. The stabilizer extract solution was collected, diluted with an HPLC eluent (acetonitrile: ultra pure water = 70: 30 v / v) as a measurement sample, and the amount of stabilizer retained was measured. The amount of sugar retained according to each concentration of stabilizer was detected. Table 1 below shows the amount of sugar retained on the surface of the equipment as a result of these.
続いて、作製した培養用生成物上に超純水を添加し、２４時間静置することで表面吸着iMatrix-511を抽出した。iMatrix-511抽出溶液を回収し、HPLC溶離液（アセトニトリル：超純水＝70：30 v/v）で希釈し測定試料とし、iMatrix-511の保持量を測定した。安定化剤の有無や濃度はiMatrix-511の保持量に影響を与えないことが示された。これらの結果である器材表面のiMatrix-511の保持量を以下の表２に示す。 Subsequently, ultrapure water was added onto the prepared culture product, and the surface adsorbed iMatrix-511 was extracted by allowing to stand for 24 hours. The iMatrix-511 extract solution was collected, diluted with an HPLC eluent (acetonitrile: ultra pure water = 70: 30 v / v) to obtain a measurement sample, and the retained amount of iMatrix-511 was measured. It was shown that the presence and concentration of stabilizers did not affect the amount of iMatrix-511 retained. Table 2 below shows the retention amount of iMatrix-511 on the surface of the equipment as a result of these.
以上順次説明した実施例１で作製した培養用生成物である器材と、比較例１、２で作製した器材それぞれを用いてiPS細胞を培養した。iPS細胞の培養では細胞を1,400細胞/ cm2播種後、毎日培地交換を行った。1週間培養後の生細胞数を比較した。 IPS cells were cultured using the equipment that is the product for culture prepared in Example 1 described above and the equipment prepared in Comparative Examples 1 and 2. In iPS cell culture, the cells were seeded at 1,400 cells / cm 2 and the medium was changed every day. The number of viable cells after one week of culture was compared.
その結果のグラフを図１に示す。図１は、培養使用直前にラミニンをコートした乾燥無表表面と、本実施例の細胞培養用生成物の表面とで、iPS細胞を培養後の細胞数比率データの比較を示すグラフである。同図で明らかなように、安定化剤無のラミニンのみをコーティングし乾燥させた器材（上述の比較例１に対応）よりも、本実施例の生成物の方が、生細胞数は多いことがわかった。また、使用直前にラミニンをコーティングした乾燥無の器材（上述の比較例２に対応）と同等の生細胞数が培養可能であった。 The resulting graph is shown in FIG. FIG. 1 is a graph showing a comparison of cell number ratio data after culturing iPS cells on a dry surfaceless surface coated with laminin just before use in culture and the surface of the cell culture product of this example. As is apparent from the figure, the product of this example has a larger number of viable cells than the device coated with laminin alone without the stabilizer and dried (corresponding to Comparative Example 1 above). I understood. In addition, the number of viable cells equivalent to laminin-coated equipment (corresponding to Comparative Example 2 described above) coated with laminin immediately before use could be cultured.
以上説明したように、本実施例の培養用生成物により、細胞外マトリックス由来基質保持表面の乾燥保存が可能であり、培養使用直前の細胞外マトリックス由来基質のコーティング作業を省略可能であり、細胞培養開始時の時間と作業工程を短縮可能となる。 As described above, the culture product of this example can be used for dry storage of the extracellular matrix-derived substrate holding surface, and the coating operation of the extracellular matrix-derived substrate immediately before the culture use can be omitted. The time and work process at the start of culture can be shortened.
なお、本発明は上記した実施例に限定されるものではなく、様々な変形例が含まれる。例えば、上記した実施例は本発明のより良い理解のために詳細に説明したのであり、必ずしも説明の全ての構成を備えるものに限定されものではない。また、ある実施例の構成の一部を他の実施例の構成に置き換えることが可能であり、また、ある実施例の構成に他の実施例の構成を加えることが可能である。また、各実施例の構成の一部について、他の構成の追加・削除・置換をすることが可能である。 In addition, this invention is not limited to an above-described Example, Various modifications are included. For example, the above-described embodiments have been described in detail for better understanding of the present invention, and are not necessarily limited to those having all the configurations described. Further, a part of the configuration of one embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of one embodiment. Further, it is possible to add, delete, and replace other configurations for a part of the configuration of each embodiment.
本発明により、簡便に利用できる接着細胞培養表面が提供でき、iPS細胞などの接着細胞を簡単に培養することができるので、再生医療や創薬の分野で極めて有用である。 According to the present invention, an adherent cell culture surface that can be easily used can be provided, and adherent cells such as iPS cells can be cultured easily, which is extremely useful in the fields of regenerative medicine and drug discovery.
Claims (15)
前記培養用表面は、細胞外マトリックス由来基質と、マルトース或いはラクトースの少なくとも1つ含む糖類を含む安定化剤を有する、
ことを特徴とする培養用生成物。 A culture product having a cell culture surface,
The culture surface has an extracellular matrix-derived substrate and a stabilizer containing a saccharide containing at least one of maltose or lactose.
A culture product characterized by the above.
前記細胞外マトリックス由来基質はラミニン、コラーゲン、あるいは接着タンパク質を少なくとも１つ含む、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The extracellular matrix-derived matrix comprises at least one laminin, collagen, or adhesion protein;
A culture product characterized by the above.
前記細胞外マトリックス由来基質はラミニンを含む、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The extracellular matrix-derived matrix comprises laminin;
A culture product characterized by the above.
前記糖類は、単糖、多糖類、あるいは前記マルトース或いはラクトース以外の二糖類を更に含む、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The saccharide further comprises a monosaccharide, a polysaccharide, or a disaccharide other than the maltose or lactose,
A culture product characterized by the above.
前記安定化剤の濃度は0.008〜0.3 mg / cm2 である、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The concentration of the stabilizing agent is 0.008~0.3 mg / cm 2,
A culture product characterized by the above.
前記安定化剤の濃度は0.01〜0.2 mg / cm2 である、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The concentration of the stabilizing agent is 0.01~0.2 mg / cm 2,
A culture product characterized by the above.
前記安定化剤の濃度は0.03〜0.1 mg / cm2 である、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The concentration of the stabilizing agent is 0.03~0.1 mg / cm 2,
A culture product characterized by the above.
前記細胞は接着細胞である、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The cell is an adherent cell;
A culture product characterized by the above.
前記細胞は人工多能性幹細胞（iPS細胞）を含む、
ことを特徴とする培養用生成物。 A culture product according to claim 1, comprising:
The cells include induced pluripotent stem cells (iPS cells),
A culture product characterized by the above.
前記細胞は人工多能性幹細胞（iPS細胞）であり、
前記安定化剤の濃度は0.03〜0.1 mg / cm2 である、
ことを特徴とする培養用生成物。 A culture product according to claim 3,
The cell is an induced pluripotent stem cell (iPS cell),
The concentration of the stabilizing agent is 0.03~0.1 mg / cm 2,
A culture product characterized by the above.
容器本体と、
前記容器本体の表面に塗られた、細胞の培養用生成物と、を備え、
前記培養用生成物は、細胞外マトリックス由来基質と、マルトース或いはラクトースの少なくとも1つ含む糖類を含む安定化剤を有する、
ことを特徴とする培養容器。 A culture vessel,
A container body;
A cell culture product applied to the surface of the container body,
The culture product has an extracellular matrix-derived substrate and a stabilizer containing a saccharide containing at least one of maltose or lactose.
A culture vessel characterized by that.
前記細胞外マトリックス由来基質はラミニンを含む、
ことを特徴とする培養容器。 The culture container according to claim 11,
The extracellular matrix-derived matrix comprises laminin;
A culture vessel characterized by that.
前記糖類は、単糖、多糖類、あるいは前記マルトース或いはラクトース以外の二糖類を更に含む、
ことを特徴とする培養容器。 The culture container according to claim 11,
The saccharide further comprises a monosaccharide, a polysaccharide, or a disaccharide other than the maltose or lactose,
A culture vessel characterized by that.
前記細胞は人工多能性幹細胞（iPS細胞）である、
ことを特徴とする培養容器。 The culture container according to claim 11,
The cell is an induced pluripotent stem cell (iPS cell),
A culture vessel characterized by that.
前記安定化剤の濃度は0.03〜0.1 mg / cm2 である、
ことを特徴とする培養容器。 The culture container according to claim 11,
The concentration of the stabilizing agent is 0.03~0.1 mg / cm 2,
A culture vessel characterized by that.
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