JP6749961B2 - 高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法 - Google Patents
高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法 Download PDFInfo
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Description
Claims (9)
- 40%を超えるコロニー形成効率及び20時間の平均細胞倍増時間である、高い増殖能力のある多能性間葉系幹細胞を識別する方法であって、
収集された間葉系幹細胞におけるNG2の発現を測定するステップと、
NG2を高発現する間葉系幹細胞を選択するステップであって、前記NG2の高発現は間葉系幹細胞の表面上のNG2量であり、且つ、前記NG2量は、ABC値として細胞1個あたり少なくとも100,000個の抗体分子である、ステップと、
を含む、方法。 - 以下の追加のステップ:
老化細胞のためのマーカーを前記間葉系幹細胞に付加することによって、請求項1において選択される前記間葉系幹細胞中の老化細胞を最小限に抑えるステップと、
老化細胞を示す前記マーカーを測定するステップと、
NG2の高い発現と老化細胞のための前記マーカーの低い表示とを有する細胞を選択するステップと、
を含む、請求項1に記載の高い増殖能力のある多能性間葉系幹細胞を識別する方法。 - 以下の追加のステップ:
系統拘束された細胞のためのマーカーを前記間葉系幹細胞に付加することによって、請求項1において選択される前記間葉系幹細胞中の系統拘束された細胞を最小限に抑えるステップと、
系統拘束された細胞を示す前記マーカーを測定するステップと、
NG2の高い発現と系統拘束された細胞のための前記マーカーの低い表示とを有する前記細胞を選択するステップと、
を含む、請求項1に記載の高い増殖能力のある多能性間葉系幹細胞を識別する方法。 - 40%を超えるコロニー形成効率及び20時間の平均細胞倍増時間である、高い増殖能力のある多能性間葉系幹細胞を識別する方法であって、
NG2と結合する基質で膜をコーティングするステップと、
前記膜の片側に収集された間葉系幹細胞を配置するステップと、
前記膜の反対側に、細胞移動を促進する走化剤を配置するステップと、
前記間葉系幹細胞を、前記膜を通って移動することを可能にするステップと、
前記膜を通ってより迅速に移動する前記間葉系幹細胞を収集するステップと、
を含む方法。 - 40%を超えるコロニー形成効率及び20時間の平均細胞倍増時間である、高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法であって、
NG2の高い発現について、収集された間葉系幹細胞を免疫表現型検査するステップと、
NG2を高発現する細胞を選択するステップであって、前記NG2の高発現は間葉系幹細胞の表面上のNG2量であり、且つ、前記NG2量は、ABC値として細胞1個あたり少なくとも100,000個の抗体分子である、ステップと、
を含む、方法。 - 以下の追加のステップ:
請求項5において選択される前記細胞を培養するステップ、
を含む、請求項5に記載の高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法。 - 以下の追加のステップ:
培養した細胞を凍結保存するステップ、
を含む、請求項6に記載の高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法。 - 以下の追加のステップ:
請求項5において選択される前記細胞を培養するステップと、
NG2の存在を監視するステップと、
を含む、請求項5に記載の高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法。 - 以下の追加のステップ:
請求項5において選択される前記細胞を培養するステップと、
NG2の存在を監視するステップと、
NG2を高発現する前記細胞を選択するステップと、
選択された前記細胞をさらに培養するステップと、
を含む、請求項5に記載の高い増殖能を有する多能性間葉系幹細胞の識別および培養のための方法。
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