JP2020507068A - 生体組織サイズを調整するための組成物、及び前記組成物を使用して生体組織のサイズを調整する方法 - Google Patents
生体組織サイズを調整するための組成物、及び前記組成物を使用して生体組織のサイズを調整する方法 Download PDFInfo
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Abstract
Description
1.発明の分野
本発明は、生体組織サイズを調整するための組成物、及び前記組成物を使用して生体組織のサイズを調整する方法に関する。
X線を使用した医療診断技法は、CT又はMRI等、二次元スキャニングから得られた画像の三次元観察から正確に疾患を診断することが可能な技術として開発された。光源の代わりに超音波が使用される場合も同様に、三次元画像を実現するための技法が診断に広く使用されてきた。しかし、現在までに開発された技法の大部分は、ミリメートル単位で画像を具体化するミリメートルレベルの分解能を有する。三次元において細胞レベルで解析され得るマイクロレベルを測定する技術は、相対的に不十分である。細胞レベル解析の大部分は、伝統的な二次元技法によって行われる。即ち、生検組織又は剖検組織等、生体組織試料が固定液で固定され、パラフィン又はポリマーに包埋され、次いで光又は電磁波が通過できるように、マイクロメートル又はナノメートルの厚さの切片が包埋組織から調製される。次に、光学又は電子顕微鏡を使用して透過画像を取得する技法を用いて、微細構造が解析される。
R1及びR2は独立して、C1〜10直鎖状又は分枝状アルキルであり、
p、q及びrは独立して、0〜10の整数である。
R1及びR2は独立して、C1〜10直鎖状又は分枝状アルキルであり、
p、q及びrは独立して、0〜10の整数である。
R1及びR2は独立して、C1〜5直鎖状又は分枝状アルキルであり、
p、q及びrは独立して、0〜5の整数である。
R1及びR2は、メチルであり、
p、q及びrは、1の整数である。
実験例1:ガラス化された生体組織サイズの変化
次の実験を行って、本発明に係る組成物が、ガラス化された組織のサイズを変化させることができるか調査した。本明細書における動物試験法は全て、韓国毒性学研究所施設内動物管理使用委員会(Institutional Animal Care and Use Committee, Korea Institute of Toxicology)のガイドライン(承認番号RS17003)に従って行った。
先ず、成体マウス(8週目)を吸入麻酔薬であるイソフルラン(1cc/分)で麻酔し、続いて段階的に50mLの冷1×PBS及び冷4%PFA(パラホルムアルデヒド)で灌流した。次いで、臓器を摘出し、4%PFA溶液に浸し、続いて4℃で24時間インキュベーションを行った。その際、上述の温度は、特異的な範囲に限定されなかったが、好ましくは0〜10℃であった。
ステップ1において調製されたガラス化された試料を、40w/v%のCHAPS、40w/v%の尿素及び50mM塩化ナトリウムを含む混合溶液において35℃で220rpmにて24時間インキュベートした。全体的脳サイズは、嗅球の部分を除いて0.8cm×1cm(幅×長さ)であった。結果を図6(a)及び図6(b)に示す。
実験例2:サイズが低下したガラス化された組織の顕微鏡画像を得ること
試料が、顕微鏡下での観察に適したサイズに変化されるように、ガラス化された組織のサイズは、本発明の組成物を使用することにより低下された。サイズが低下したガラス化された脳組織を、1×対物レンズを使用した顕微鏡下で観察し、そこから、マウス脳の緑色蛍光タンパク質(GFP)が確認された(図6(c))。
Claims (9)
- 生体組織サイズを調整するための組成物であって、下記の式1によって表される化合物、その光学異性体、その水和物又はその塩、及びアルカリ金属ハロゲン化物を含む組成物
R1及びR2は独立して、C1〜10直鎖状又は分枝状アルキルであり、
p、q及びrは独立して、0〜10の整数である)。 - R1及びR2が独立して、C1〜5直鎖状又は分枝状アルキルであり、p、q及びrが独立して、0〜5の整数である、請求項1に記載の生体組織サイズを調整するための組成物。
- R1及びR2が独立して、メチルであり、p、q及びrが独立して、1の整数である、請求項1に記載の生体組織サイズを調整するための組成物。
- 式1によって表される化合物が、下記の式2によって表される化合物又はその水和物である、請求項1に記載の生体組織サイズを調整するための組成物。
- 尿素、CHAPSO(3-([3-コラミドプロピル]ジメチルアンモニオ)-2-ヒドロキシ-1-プロパンスルホナート)、スクロース、フルクトース、グリセロール、ジアトリゾ酸、トリトンX-100、ツイーン-20、2,2'-チオジエタノール、イオヘキソール及び抱水クロラールからなる群から選択される1種以上の化合物をさらに含む、請求項1に記載の生体組織サイズを調整するための組成物。
- 模擬体液をさらに含み、式1によって表される化合物、その光学異性体、その水和物又はその塩の濃度が、30〜60w/v%であり、アルカリ金属ハロゲン化物の濃度が、1〜5w/v%である、請求項1に記載の生体組織サイズを調整するための組成物。
- 生体組織が、脳、血管、肝臓、肺、腎臓、膵臓又は腸である、請求項1に記載の生体組織サイズを調整するための組成物。
- 生体組織のサイズを調整する方法であって、生体組織を請求項1に記載の組成物と接触させることにより、組織のサイズを調整するステップを含む、方法。
- 10℃〜50℃の温度範囲で行われる、請求項8に記載の生体組織のサイズを調整する方法。
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KR101849698B1 (ko) * | 2017-08-24 | 2018-04-18 | 한국화학연구원 | 생체 조직 크기 조절용 조성물 및 상기 조성물을 이용한 생체 조직의 크기 조절 방법 |
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JP6964674B2 (ja) | 2021-11-10 |
US20200140479A1 (en) | 2020-05-07 |
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