ヒトES細胞、ヒトiPS細胞剥離液 (CTK溶液)

　Dissociation Solution
　　for human ES/iPS Cells

【特徴】

剥離液を加えピペッティング操作するだけで継代に適した大きさのコロニーに分割できるため、カッター等でコロニーを裁断して傷つけることはありません。
高い生存率で継代できます。
オンフィーダー、フィーダーレスの両方に使用できます。
全ロットの品質試験を行っておりますので、安心してお使いいただけます。

※保存温度：-20℃以下
※特許 4317337号

概要

リプロセルの剥離液は、京都大学中辻憲夫教授 (iCeMS)・末盛博文准教授 (再生医科学研究所) により開発されたものであり一般的にCTK溶液と呼ばれています。このCTK溶液の特許 (権利化済み) はリプロセルが保有しており、「Dissociation Solution for human ES/iPS Cells」として販売しております。また、CTK溶液はヒトiPS細胞の剥離液のゴールドスタンダードとしても広く使われています。

ヒトES細胞やヒトiPS細胞を継代するときには、single cell suspensionにせずコロニーを維持したままで行うことが重要です。例えば、一般の細胞株を剥離する際に使用されるトリプシン-EDTAなどを使用してヒトES細胞およびサルES細胞を継代した場合、生存率は数パーセントと著しく低下します。

これに対して、当社の剥離液は、添加するだけで図に示すように継代に適した大きさのコロニーを調製することが可能です。このことにより、継代時の細胞の生存率がTrypsin-EDTA溶液に比べ約10倍と高いのが特徴です。また、カニクイザルES細胞やヒトES細胞で、3年以上の染色体異常のない培養実績があります。

【動画】細胞剥離液 (RCHETP002) での細胞の剥離

製品リスト

品番	製品名
RCHETP002	Dissociation solution for human ES/iPS cells（30 mL）

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プロトコル

Dissociation Solution for human ES/iPS Cells (CTK溶液) プロトコル

操作動画

1.ヒトiPS細胞の形態

iPS細胞の形態について解説します。細胞の状態の判断にご活用ください。良い状態のiPS細胞は、コロニーの輪郭が明瞭で内部の細胞密度が高くなっています。

2.フィーダー細胞の播種方法

iPS細胞の培養に使用するフィーダー細胞は、その播種密度が重要になります。3×10⁴ 細胞/cm²が目安になります。

3.ヒトiPS細胞の継代方法

Dissociation Solution for human ES/iPS Cells (CTK溶液) を用いることで、高生存率かつ簡便な継代が可能になります。ピペッティング操作だけで、適度な大きさのコロニーにほぐれ、そのまま継代できます。通常3－4日に1回、継代を行います。

4.細胞の凍結保存方法と融解方法

Freezing Medium for human ES/iPS Cellsを使用し、ガラス化法で凍結することで、高い生存率での凍結保存が可能になります。ガラス化法とは、細胞を液体窒素で急速凍結する方法で、細胞へのダメージを最小限に抑えます。また、融解時も、温めておいた培養液を凍結細胞に直接加えることで急速解凍を行います。凍結・融解の作業に時間がかかると、生存率が低下するのでご注意ください。

文献・発表

本製品を実験にご使用いただいた論文のリストはこちらです。

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