JP5846550B2 - 短繊維足場材料、短繊維−細胞複合凝集塊作製方法及び短繊維−細胞複合凝集塊 - Google Patents
短繊維足場材料、短繊維−細胞複合凝集塊作製方法及び短繊維−細胞複合凝集塊 Download PDFInfo
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Description
また、前記ポリマーブラシを親水性の材料で形成してよい。
また、短繊維を中空状としてよい。
また、短繊維は電界紡糸繊維を切断したものであってよい。
本発明の他の側面によれば、上述の短繊維足場材料及び細胞を分散させた培養液中で培養を行うことにより、前記短繊維足場材料及び細胞を含む短繊維−細胞複合凝集塊を形成する、短繊維−細胞複合凝集塊作製方法が与えられる。
ここで、前記培養液は血清と成長因子の少なくとも何れかを含んでよい。
また、培養中に外部場を印加することにより前記短繊維に配向性を持たせてよい。
本発明の更に他の側面によれば、上述の短繊維足場材料と細胞とが凝集した、多孔質の短繊維−細胞複合凝集塊が与えられる。
ここで、表面と内部との間の循環系を提供する、径が8nm以上の経路が形成されていてよい。
また、前記短繊維が配向性を有してよい。
まず、図1(a)に示す反応により、原始移動ラジカル重合(ATRP)の開始基を有する4-ビニルベンゼン−2-ブロモプロピオネート(4-vinylbenzyl-2-bromopropionate)(VBP)とスチレン(ST)のランダム共重合を行った。得られた共重合体であるポリ(ST−r−VBP)(Mn=105200、Mw/Mn=2.82)を用いて電界紡糸を行い、直径593nm±74nmの電界紡糸ファイバー(不織布)を得た。このようにして得られた電界紡糸ファイバーのSEM像を図2(a)に示す。
上述のようにして作製した短繊維足場材料を使用し、以下のようにしてこの足場材料とヒヨコ角膜実質細胞とから構成された細胞組織を得た。
得られた短繊維を消毒用のアルコールを用いて簡易的に滅菌を行った後に、遠心操作によりアルコールを除去し、その後24時間リン酸化緩衝液で十分に洗浄し残存アルコールを除去した。遠心沈降操作の後、クリーベン内において滅菌状態を維持した状態でリン酸緩衝溶液を除去し、その後、10%血清を含むEagle MEM培地を加え、さらに2時間ほど分散撹拌することで、短繊維懸濁培地を得た。短繊維が約5mg/mlとなるように調整した短繊維懸濁培地3mlに100,000細胞/mlの濃度の細胞分散液3mlを加え、十分にピペッティングすることにより細胞と短繊維の分散を行った。その後、この混合溶液を細胞培養用シャーレに播種し、炭酸ガスインキュベーター内で37度、48時間培養を行ない短繊維−細胞複合凝集塊を得た。
Claims (10)
- 平均長が10μm〜500μmであり、かつ、平均直径が200nm〜4μmである紡糸ファイバーからなり、前記紡糸ファイバーの表面にポリマーブラシが形成されている、培養液中に添加することにより細胞とともに凝集する短繊維足場材料。
- 前記ポリマーブラシは親水性の材料で形成された、請求項1に記載の短繊維足場材料。
- 中空状である、請求項1または2に記載の短繊維足場材料。
- 電界紡糸繊維を切断した、請求項1から3の何れかに記載の短繊維足場材料。
- 請求項1から4の何れかに記載の短繊維足場材料及び細胞を分散させた培養液中で培養を行うことにより、前記短繊維足場材料及び細胞を含む短繊維−細胞複合凝集塊を形成する、短繊維−細胞複合凝集塊作製方法。
- 前記培養液は血清と成長因子の少なくとも何れかを含む、請求項5に記載の短繊維−細胞複合凝集塊作製方法。
- 培養中に外部場を印加することにより前記短繊維に配向性を持たせる、請求項6に記載の短繊維−細胞複合凝集塊作製方法。
- 請求項1から4の何れかに記載の短繊維足場材料と細胞とが凝集した、多孔質の短繊維−細胞複合凝集塊。
- 表面と内部との間の循環系を提供する、径が8nm以上の経路が形成されている、請求項8に記載の短繊維−細胞複合凝集塊。
- 前記短繊維が配向性を有する、請求項8または9に記載の短繊維−細胞複合凝集塊。
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JP2015159778A (ja) * | 2014-02-28 | 2015-09-07 | 日本バイリーン株式会社 | 光学的測定用細胞培養担体、光学的測定用ウェルプレート、及び細胞の光学的測定方法 |
JP6115842B1 (ja) * | 2016-02-17 | 2017-04-19 | 公立大学法人横浜市立大学 | 足場材料への細胞播種方法及びその装置 |
JP6799316B2 (ja) * | 2016-09-12 | 2020-12-16 | 国立研究開発法人物質・材料研究機構 | 配向性を有する短繊維−細胞複合凝集塊を製造するための方法 |
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SE454518B (sv) * | 1984-05-21 | 1988-05-09 | Statens Bakteriologiska Lab | Forfarande for odling av diploida celler i nervaro av cellulosafibrer |
ATE464408T1 (de) * | 2003-02-13 | 2010-04-15 | Teijin Ltd | Poröse faser, poröses fasergebilde und herstellungsverfahren |
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