JP2023054120A - オルガノイドの生体移植用組成物 - Google Patents
オルガノイドの生体移植用組成物 Download PDFInfo
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Abstract
Description
ゼラチン、コラーゲン、フィブリン糊、またはこれらの組み合わせを含む生体移植用組成物を提供し、
好ましくは、前記オルガノイドは、腸オルガノイド、網膜オルガノイド、腎臓オルガノイド、肝臓オルガノイド、胃オルガノイド、前立腺オルガノイド、乳房オルガノイド、内耳オルガノイド、心臓筋線維オルガノイド、肝臓内皮オルガノイド、膵臓オルガノイド、卵管オルガノイド及び大脳オルガノイドからなる群から選択され、および/または、
前記ゼラチンは、組成物の総重量に対して2.5~10%(w/v)含まれ、および/または、
前記コラーゲンは、組成物の総重量に対して10~20%(v/v)含まれ、および/または、
前記フィブリン糊は、組成物の総重量に対して10~15%(v/v)含まれる。
その混合物を個体に投与する工程、を含むオルガノイドの移植方法を提供する。
EGFPマウスから大腸組織を分離して、酵素を使用して大腸クリプト(crypt)を分離し、マトリゲルと大腸オルガノイド用培地を1:1で混ぜて、コーティングされていない48ウェルプレートに接種した。これをインキュベーターに入れて、20分後にマトリゲルが固まったことを確認し、大腸オルガノイド用培地を添加して5日間培養し、下記に使用される生体移植用大腸オルガノイドを製作した。
1.1. 大腸組織損傷モデルの製作
オルガノイドを製作する大腸組織損傷モデルを下記のように製造した。大腸オルガノイドを移植する野生型マウスを0.5M EDTAに5分間露出させ、電動歯ブラシで2分間大腸内壁のクリプトが除去されるように物理的な損傷を加えた。
前記で製作した大腸オルガノイドがGFPを発現するように、大腸オルガノイド(colon organoid)を準備した。すべての大腸オルガノイドは、移植1日前に10μMのY-27632をオルガノイド培地に処理して生存率を極大化した。また、セルリカバリーソリューション(Cell recovery solution)を処理して培養時に使用したマトリックスを完全に除去した。
前記実施例1で移植した大腸オルガノイドのスキャフォールドに係る移植率を評価するために、下記のように行った。
前記実施例1で移植した大腸オルガノイドのスキャフォールドに係る生存率及び移植効率を評価するために、下記のように行った。
前記実施例1で移植した大腸オルガノイドの安定性を評価するために、移植した後7日目に解剖検査して、解剖検査組織の形態及び病変発生の有無を確認した。
前記実施例1で移植した大腸オルガノイドから正常なオルガノイドが形成されるか否かを評価するために、移植された大腸組織から2回目の大腸オルガノイド(2次オルガノイド)を形成させた。
Claims (3)
- 腸オルガノイドとフィブリン糊とを含む生体移植用組成物。
- 前記フィブリン糊は、組成物の総重量に対して10~15%(v/v)の量で含まれる請求項1に記載の生体移植用組成物。
- 請求項1に記載の組成物を、ヒトを除く個体に投与する工程、を含む腸オルガノイドの移植方法。
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020180152804A KR20200065892A (ko) | 2018-11-30 | 2018-11-30 | 오가노이드의 생체 이식용 조성물 |
KR10-2018-0152804 | 2018-11-30 | ||
PCT/KR2019/013527 WO2020111507A1 (ko) | 2018-11-30 | 2019-10-15 | 오가노이드의 생체 이식용 조성물 |
JP2021531269A JP2022510982A (ja) | 2018-11-30 | 2019-10-15 | オルガノイドの生体移植用組成物 |
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KR20220068172A (ko) | 2020-11-17 | 2022-05-25 | 연세대학교 산학협력단 | 식도 오가노이드 배양 및 이식을 위한 탈세포 식도 조직 유래 지지체 및 이의 제조방법 |
KR20220068173A (ko) | 2020-11-17 | 2022-05-25 | 연세대학교 산학협력단 | 췌장 오가노이드 배양 및 이식을 위한 탈세포 췌장 조직 유래 지지체 및 이의 제조방법 |
CN114317413A (zh) * | 2022-01-17 | 2022-04-12 | 中山大学 | 肠道类器官的培养方法及其在肠道毒性测试中的应用 |
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AU2002367580A1 (en) * | 2001-05-16 | 2003-09-22 | Tracy C. Grikscheit | Tissue-engineered organs |
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GB201111244D0 (en) * | 2011-06-30 | 2011-08-17 | Konink Nl Akademie Van Wetenschappen Knaw | Culture media for stem cells |
CN102198022B (zh) * | 2011-05-23 | 2013-04-17 | 西安交通大学 | 一种活性细胞-水凝胶类器官结构体的立体成形方法 |
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WO2014066649A1 (en) * | 2012-10-26 | 2014-05-01 | The Regents Of The University Of California | A strategy for engineering various 3d tissues, organoids and vasculature |
HRP20240170T1 (hr) * | 2013-05-08 | 2024-04-12 | Prokidney | Organoidi koji sadrže izolirane renalne stanice i njihove uporabe |
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WO2017036533A1 (en) * | 2015-09-03 | 2017-03-09 | Ecole Polytechnique Federale De Lausanne (Epfl) | Three-dimensional hydrogels for culturing adult epithelial stem cells and organoids |
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GB201610748D0 (en) | 2016-06-20 | 2016-08-03 | Koninklijke Nederlandse Akademie Van Wetenschappen | Improved diffrentation method |
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US20220226538A1 (en) | 2022-07-21 |
JP2022510982A (ja) | 2022-01-28 |
WO2020111507A1 (ko) | 2020-06-04 |
CN113660962A (zh) | 2021-11-16 |
KR102597594B1 (ko) | 2023-11-02 |
EP3888715A4 (en) | 2022-08-17 |
KR20210069619A (ko) | 2021-06-11 |
JP7542880B2 (ja) | 2024-09-02 |
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