JP6854471B2 - 神経細胞の密集体 - Google Patents
神経細胞の密集体 Download PDFInfo
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Description
(1)control PBS (Phosphate Buffered Saline) washing>>>culture medium
(2)H2O2 3hour treatment>>>PBS washing>>>culture medium
(3)H2O2 3hour treatment>>>PBS washing>>>culture medium containing Edaravone
処理を行ってから2日後に、マーカーであるTau 1 で免疫染色した後、評価を行った。なお、評価項目は、Directionality:軸索の方向性である。具体的には、図22内のオブジェクトの方向性を計測した。この場合、0度が流路(チャネル13)方向である。軸索は、束になっていて同じ方向に延びているので、チャネル13内で、例えば、図22(b)に示されるように、酸化ストレスを付与したときや、図22(c)に示されるように、酸化ストレスと、神経疾患の薬剤として用いられる抗酸化剤(Edaravone )とを同時に付与したときの形態変化を比較することができる。そして、図23に示されるような評価実験の結果は、酸化ストレスを付与すると、軸索方向に伸展する軸索が変性するので、流路方向(0度)への方向性が減少し複雑な形状に変化することを示している。形態変化の評価として、図23に示されるオブジェクトの配向性(Directionality)を調べると、負荷をかけていないnegative controlの軸索束の配向性が高く、次に、薬剤及びストレスを付与したとき、その次に、酸化ストレスだけを付与したとき、という順で、配向性の差を観ることができた。これにより、神経細胞が受けたストレスを、軸索の形態変化として、画像処理に基づいて評価することができることが分かる。
11 モジュール
12a 第1チャンバー
12b 第2チャンバー
13 チャネル
15 基板
16 上板
17 シール部材
18 培養液
21 カッター
22 ピペット
Claims (5)
- ヒトiPS細胞由来の細胞体と該細胞体から延びた軸索とを有する複数の神経細胞から成る神経細胞の密集体であって、
該神経細胞の密集体の各々は、1つのスフェロイドと、該スフェロイドから1方向に延びる長さ1〔mm〕以上の1つの軸索束とを有し、他の神経細胞の密集体及び軸索培養のための支持体から分離された状態で存在し、
前記スフェロイドは複数の細胞体の密集体であり、
前記軸索束は複数の軸索の束であって、前記軸索束内では、ウェスタンブロッティングによる細胞体の存在を示すマーカーがネガティブであることに相当する程度に細胞体が不存在な状態を示す、
神経細胞の密集体。 - 前記軸索束が4.5〔mm〕以上の長さを有する請求項1に記載の神経細胞の密集体。
- 前記スフェロイドがグリア細胞を含む請求項1又は2に記載の神経細胞の密集体。
- 前記神経細胞の密集体は運動神経細胞である請求項1〜3のいずれか1項に記載の神経細胞の密集体。
- 前記軸索束は軸索末端において骨格筋細胞と接合している状態である請求項4に記載の神経細胞の密集体。
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Application Number | Priority Date | Filing Date | Title |
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US201662328905P | 2016-04-28 | 2016-04-28 | |
US62/328,905 | 2016-04-28 |
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US20200386742A1 (en) * | 2017-12-04 | 2020-12-10 | The Administrators Of The Tulane Educational Fund | Cell systems using spheroids and methods of making and using the same |
EP3995509A4 (en) | 2019-07-05 | 2023-07-19 | Jiksak Bioengineering, Inc. | METHOD OF INDUCING NEURAL SYNAPSIS FORMATION AND MICROBEADS USED IN THIS METHOD |
EP4006134A4 (en) * | 2019-07-22 | 2023-10-04 | Jiksak Bioengineering, Inc. | CULTURE METHOD AND CULTURE CONTAINER |
CN111996114B (zh) * | 2019-10-12 | 2023-05-12 | 南方科技大学 | 一种用于神经细胞轴突分离培养的多室型微流控装置 |
JP2022051245A (ja) * | 2020-09-18 | 2022-03-31 | 国立大学法人 筑波大学 | 神経束および神経束の製造方法 |
WO2022059793A1 (ja) * | 2020-09-18 | 2022-03-24 | 国立大学法人 筑波大学 | 神経束および神経束の製造方法 |
US20230390459A1 (en) * | 2020-10-29 | 2023-12-07 | Keio University | Artificial axon bundle |
WO2022154080A1 (ja) * | 2021-01-15 | 2022-07-21 | 国立大学法人東京大学 | 複雑な活動と可塑性を示すインビトロ神経回路組織、その神経活動の観察装置およびこれを用いた物質のスクリーニング方法 |
WO2022261775A1 (en) * | 2021-06-18 | 2022-12-22 | 9493662 Canada Inc. | Device and method for preparing compartmentalized in vitro models with an elongated component of a biological material |
JP2023005361A (ja) | 2021-06-29 | 2023-01-18 | 学校法人東北工業大学 | 薬剤の影響予測システム |
WO2023173215A1 (en) * | 2022-03-16 | 2023-09-21 | Ananda Devices Inc. | Devices and methods for preparing in vitro models of neuromuscular junctions |
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JP7151966B2 (ja) | 2017-06-14 | 2022-10-12 | 一般財団法人生産技術研究奨励会 | 神経細胞を培養する装置、神経細胞を培養する方法、軸索束の形態変性を配向性解析によって数値化する方法、神経組織、軸索束内のプロテインを解析及び同定する方法並びに神経細胞の使用方法 |
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