JP2022017508A - 凍結保存組成物およびその使用方法 - Google Patents
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- JP2022017508A JP2022017508A JP2021181056A JP2021181056A JP2022017508A JP 2022017508 A JP2022017508 A JP 2022017508A JP 2021181056 A JP2021181056 A JP 2021181056A JP 2021181056 A JP2021181056 A JP 2021181056A JP 2022017508 A JP2022017508 A JP 2022017508A
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Abstract
Description
-80℃の温度への低速凍結保存法に適する凍結保存組成物は、ダルベッコ修飾イーグル培地(DMEM)中で混合した7.5%v/vのカルボキシル化ポリリシンおよび5%w/vのデキストラン-40を含む。コハク酸無水物との反応によりポリリシンのアミノ基のおよそ60%をカルボキシル化した。その溶媒としての水中のカルボキシル化ポリリシンを、DMEM中1:3の比でデキストラン-40と混合する。
-80℃の温度への低速凍結保存法に適する凍結保存組成物は、DMEM中で混合した7.5%v/vのカルボキシル化ポリリシンおよび5%w/vのエクトインを含む。コハク酸無水物との反応によりポリリシンのアミノ基のおよそ60%をカルボキシル化した。その溶媒としての水中のカルボキシル化ポリリシを、DMEM中1:3の比でエクトインと混合する。
-80℃の温度への低速凍結保存法に適する凍結保存組成物は、DMEM中で混合した7.5%v/vのカルボキシル化ポリリシン、5%w/vのエクトインおよび5%w/vのデキストラン-40を含む。コハク酸無水物との反応によりポリリシンのアミノ基のおよそ60%をカルボキシル化した。その溶媒としての水中のカルボキシル化ポリリシを、DMEM中1:3の比でデキストラン-40およびエクトインと混合する。
凍結保存法に適する凍結保存組成物は、DMEM中で混合した7.5%v/vのカルボキシル化ポリリシン、20%w/vのエクトインおよび20%w/vのデキストラン-40を含む。その溶媒としての水中のカルボキシル化ポリリシを、DMEM中1:3の比でデキストラン-40およびエクトインと混合する。
-100℃未満の温度へのガラス化および/または超急速凍結を含む高速凍結保存に適する凍結保存組成物は、DMEM中で混合した52.5%v/vのカルボキシル化ポリリシン、35%w/vのエクトインおよび35%w/vのデキストラン-40を含む。コハク酸無水物との反応によりポリリシンのアミノ基のおよそ60%をカルボキシル化する。その溶媒としての水中のカルボキシル化ポリリシを、DMEMを加えることによりそれらの最終濃度にデキストラン-40およびエクトインと混合する。
-100℃未満の温度へのガラス化および/または超急速凍結を含む高速凍結保存に適する凍結保存組成物は、DMEM中で混合した37.5%v/vのカルボキシル化ポリリシン、25%w/vのエクトインおよび25%w/vのデキストラン-40を含む。コハク酸無水物との反応によりポリリシンのアミノ基のおよそ60%をカルボキシル化する。その溶媒としての水中のカルボキシル化ポリリシを、DMEMを加えることによりそれらの最終濃度にデキストラン-40およびエクトインと混合する。
Claims (42)
- カルボキシル化ポリアミノ酸、有機両性剤、および場合により多糖を含む、凍結保存組成物。
- カルボキシル化ポリアミノ酸がカルボキシル化ポリリシンである、請求項1に記載の組成物。
- カルボキシル化ポリリシン上の50%超のアミノ基がブロックされる、請求項1または2のいずれか一項に記載の組成物。
- カルボキシル化ポリリシン上の約50%~約65%のアミノ基がブロックされる、請求項1~3のいずれか一項に記載の組成物。
- カルボキシル化ポリアミノ酸が凍結保存組成物の0.1%v/v~約20%v/vの範囲の量で存在する、請求項1~4のいずれか一項に記載の組成物。
- カルボキシル化ポリアミノ酸が凍結保存組成物の1.0%v/v~約10%v/vの範囲の量で存在する、請求項1~5のいずれか一項に記載の組成物。
- 有機両性剤がエクトイン、ヒドロキシエクトイン、およびそれらの組合せより選択される、請求項1~6のいずれか一項に記載の組成物。
- 有機両性剤が凍結保存組成物の約0.1%w/v~約20%w/vの範囲の量で存在する、請求項1~7のいずれか一項に記載の組成物。
- 有機両性剤が凍結保存組成物の約1.0%w/v~約10%w/vの範囲の量で存在する、請求項1~8のいずれか一項に記載の組成物。
- 多糖がデキストランである、請求項1~9のいずれか一項に記載の組成物。
- 多糖が凍結保存組成物の約0.1%w/v~約20%w/vの範囲の量で存在する、請求項1~10のいずれか一項に記載の組成物。
- 多糖が凍結保存組成物の約1.0%w/v~約10%w/vの範囲の量で存在する、請求項1~11のいずれか一項に記載の組成物。
- 多糖および有機両性剤が凍結保存組成物の約3.0%w/v~約8%w/vの範囲の同じ量で存在する、請求項1~12のいずれか一項に記載の組成物。
- 凍結保存組成物が約7.5%v/v以下のカルボキシル化ポリアミノ酸、約5%w/v以下の有機両性剤、および約5%w/v以下の多糖を含む、請求項1~13のいずれか一項に記載の組成物。
- カルボキシル化ポリアミノ酸が約7.5%v/v以下のカルボキシル化ポリリシンを含み、有機両性剤が約5%w/v以下のエクトインおよび/またはヒドロキシエクトインを含み、多糖が約5%w/v以下のデキストランを含む、請求項1~14のいずれか一項に記載の組成物。
- カルボキシル化ポリアミノ酸が約1.1:1~約3:1の範囲の有機両性剤に対する比で存在する、請求項1~15のいずれか一項に記載の組成物。
- カルボキシル化ポリアミノ酸が約1.3:1~約1.7:1の有機両性剤に対する比で存在する、請求項1~16のいずれか一項に記載の組成物。
- 有機両性剤がエクトインであり、多糖がデキストランである、請求項2に記載の組成物。
- DMEM、EMEM、X-VIVO、生理食塩水、水、およびそれらの組合せからなる群より選択される溶媒をさらに含む、請求項1~18のいずれか一項に記載の組成物。
- 生体試料を得ること;および生体試料を凍結保存組成物と接触させることを含む、生体試料を凍結保存する方法であって、該凍結保存組成物が約1%v/v~約10%v/vのカルボキシル化ポリアミノ酸、約1%w/v~約10%w/vの有機両性剤、および約0%w/v~約10%w/vの多糖を含む、方法。
- 凍結保存組成物が、約1%v/v~約10%v/vのカルボキシル化ポリリシン、ここで該カルボキシル化ポリリシンは50%超のアミノ基がブロックされており、約1%w/v~約10%w/vのエクトインおよび約1%w/v~約10%w/vのデキストランを含む、請求項20に記載の方法。
- カルボキシル化ポリリシンが約1.1:1:1~約3:1:1の範囲の有機両性剤と多糖に対する比で存在する、請求項20~21のいずれか一項に記載の方法。
- カルボキシル化ポリリシンが約1.5:1:1の有機両性剤と多糖に対する比で存在する、請求項20~22のいずれか一項に記載の方法。
- 凍結保存組成物を生体試料に加えること、および生体試料を低速凍結保存することを含む、生きた状態で生体試料を貯蔵する方法であって、該凍結保存組成物が、約1%v/v~約10%v/vのカルボキシル化ポリリシン、ここで該カルボキシル化ポリリシンは50%超のアミノ基がカルボキシル基でブロックブロックされており、約1%w/v~約10%w/vのエクトイン、および約0%w/v~約10%w/vのデキストランを含む、方法。
- カルボキシル化ポリリシンが約1.5:1:1のエクトインとデキストランに対する比で存在する、請求項24に記載の方法。
- 生体試料を解凍することをさらに含み、該生体試料が50%超の解凍後生存率を示す、請求項24~25のいずれか一項に記載の方法。
- 約20%v/v~約55%v/vのカルボキシル化ポリリシン、約20%~約55%のエクトインまたはヒドロキシエクトイン、および場合により約20%~約55%の多糖を含む、高速凍結保存のための凍結保存組成物。
- カルボキシル化ポリリシン上の50%超のアミノ基がブロックされる、請求項27に記載の組成物。
- カルボキシル化ポリリシン上の約60%のアミノ基がブロックされる、請求項27~28のいずれか一項に記載の組成物。
- カルボキシル化ポリリシンが凍結保存組成物の約22.5%v/v~約52.5%v/vの範囲の量で存在する、請求項27~29のいずれか一項に記載の組成物。
- エクトインまたはヒドロキシエクトインが凍結保存組成物の約25%w/v~約35%w/vの範囲の量で存在する、請求項27~30のいずれか一項に記載の組成物。
- 多糖がデキストランである、請求項27~31のいずれか一項に記載の組成物。
- デキストランが凍結保存組成物の約25%w/v~約35%w/vの範囲の量で存在する、請求項27~32のいずれか一項に記載の組成物。
- 多糖および有機両性剤が凍結保存組成物の約25.0%w/v~約35%w/vの範囲の同じ量で存在する、請求項27~33のいずれか一項に記載の組成物。
- カルボキシル化ポリリシンが約1.1:1~約3:1の範囲のエクトインおよび/またはヒドロキシエクトインに対する比で存在する、請求項27~34のいずれか一項に記載の組成物。
- カルボキシル化ポリリシンが約1.3:1~約1.7:1のエクトインおよび/またはヒドロキシエクトインに対する比で存在する、請求項27~35のいずれか一項に記載の組成物。
- 生体試料を得ること;および生体試料を凍結保存組成物と接触させることを含む、生体試料を凍結保存する方法であって、該凍結保存組成物が約20%v/v~約55%v/vのカルボキシル化ポリリシン、約20%w/v~約55%w/vの有機両性剤、および場合により約20%w/v~約55%w/vの多糖を含む、方法。
- 凍結保存組成物が、約20%v/v~約55%v/vのカルボキシル化ポリリシン、ここで該カルボキシル化ポリリシンは50%超のアミノ基がブロックされており、約25%w/v~約35%w/vのエクトインおよび約25%w/v~約35%w/vのデキストランを含む、請求項37に記載の方法。
- カルボキシル化ポリリシンが約1.1:1:1~約3:1:1の範囲のエクトインとデキストランに対する比で存在する、請求項37~38のいずれか一項に記載の方法。
- カルボキシル化ポリリシンが約1.5:1:1のエクトインとデキストランに対する比で存在する、請求項37~39のいずれか一項に記載の方法。
- 凍結保存組成物を生体試料に加えること、生体試料をガラス化または超急速凍結させることを含む、生きた状態で生体試料を貯蔵する方法であって、該凍結保存組成物が、約20%v/v~約55%v/vのカルボキシル化ポリリシン、ここで該カルボキシル化ポリリシンは50%超のアミノ基がブロックされており、約20%w/v~約55%w/vのエクトインおよび約20%w/v~約55%w/vのデキストランを含む、方法。
- カルボキシル化ポリリシンが約1.5:1:1からのエクトインとデキストランに対する比で存在する、請求項41に記載の方法。
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JP2012217342A (ja) * | 2011-04-04 | 2012-11-12 | Bio Verde:Kk | 多能性幹細胞その他の分散浮遊可能な細胞用の凍結保存液および凍結保存法 |
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WO2017087401A1 (en) | 2017-05-26 |
CN108882699A (zh) | 2018-11-23 |
JP7323290B2 (ja) | 2023-08-08 |
JP2018533377A (ja) | 2018-11-15 |
US20180325100A1 (en) | 2018-11-15 |
EP3376862A1 (en) | 2018-09-26 |
EP3376862A4 (en) | 2019-06-19 |
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