JP2013543725A - 培養膵島 - Google Patents
培養膵島 Download PDFInfo
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- JP2013543725A JP2013543725A JP2013535111A JP2013535111A JP2013543725A JP 2013543725 A JP2013543725 A JP 2013543725A JP 2013535111 A JP2013535111 A JP 2013535111A JP 2013535111 A JP2013535111 A JP 2013535111A JP 2013543725 A JP2013543725 A JP 2013543725A
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Abstract
Description
従来より、さまざまな臓器保存溶液が開発されてきた。
Unisol(登録商標)「メンテナンス」溶液が開発され、連続灌流によりO2の十分な供給が維持される場合、ATP予備を再合成することができる温度範囲と一致する、7〜10℃の範囲内の温度で試験された。例えば、多数の研究が、酸素供給は肝臓の低体温保存中必須であることを示した。
例えば約50〜約200μU/ml、例えば約100μU/mlのペニシリンおよび約50〜約200μg/ml、例えば約100μg/mlのストレプトマイシンを含有し、望ましくない微生物の増殖を阻害する。実施形態では、培地に、細胞組織の培養基質への付着を促進するタンパク質を含有する少量の哺乳類血清を添加することもできる。例えば、血清は、混合物の体積に対して、約5〜20体積%、例えば約15体積%の量で存在することができ、培養を行う島を含む細胞生成物に対して異種である。
Claims (15)
- 膵島を培地において外分泌物質の存在下でエクスビボ培養するステップを含む、膵島の調製方法。
- 膵組織を消化酵素で処理することにより、前記島を膵組織から単離するステップをさらに含む、請求項1に記載の方法。
- 膵島を含むドナー組織を消化酵素で処理し、島を含む細胞生成物を得るステップ;および
該島を含む細胞生成物を培地に外分泌物質の存在下で播種するステップ
を含む、パンクレアタイトの調製方法。 - 前記島が未熟な島を含む、請求項1〜3のいずれか一項に記載の方法。
- 前記培地がガス透過膜のような膜、好適にはポリカーボネートガス透過性シリコーン膜培養フラスコ上に存在する、請求項1〜4のいずれか一項に記載の方法。
- 前記島が前記培地中に存在する外分泌組織および内分泌組織の総面積の約20%より大きい量で存在し、前記島の量が前記島に含まれる面積の前記培地中に存在する外分泌組織および内分泌組織の総面積に対する比により計算され、例えば、前記島が前記培地中に存在する外分泌組織および内分泌組織の総面積の約30%より大きい量で存在するか、または前記島が前記培地中に存在する外分泌組織および内分泌組織の総面積の約40%より大きい量で存在する、請求項1〜5のいずれか一項に記載の方法。
- 前記島が前記培地中に存在する外分泌組織および内分泌組織の総面積の約70%未満の量で存在し、例えば、前記島が前記培地中に存在する外分泌組織および内分泌組織の総面積の約60%未満の量で存在する、または前記島が前記培地中に存在する外分泌組織および内分泌組織の総面積の約50%未満の量で存在する、請求項1〜6のいずれか一項に記載の方法。
- 前記外分泌物質が前記培地中に存在する外分泌組織および内分泌組織の総面積の約60%より大きい量で存在し、前記外分泌物質の量が前記外分泌物質に含まれる面積の前記培地中に存在する外分泌組織および内分泌組織の総面積に対する比により計算され、例えば、前記外分泌物質が前記培地中に存在する外分泌組織および内分泌組織の総面積の約70%より大きい量で存在する、または前記外分泌物質が前記培地中に存在する外分泌組織および内分泌組織の総面積の約80%より大きい量で存在する、請求項1〜7のいずれか一項に記載の方法。
- 前記外分泌物質が前記培地中に存在する外分泌組織および内分泌組織の総面積の約50%未満の量で存在し、前記外分泌物質の量が前記外分泌物質に含まれる面積の前記培地中に存在する外分泌組織および内分泌組織の総面積に対する比により計算され、例えば、前記外分泌物質が前記培地中に存在する外分泌組織および内分泌組織の総面積の約40%未満の量で存在するか、または前記外分泌物質が前記培地中に存在する外分泌組織および内分泌組織の総面積の約30%未満の量で存在する、請求項1〜7のいずれか一項に記載の方法。
- 前記島が未成熟ドナー、若年ドナー、哺乳類ドナー、ヒトドナー、ブタドナー、ウシドナー、および霊長類ドナーからなる群から選択されるドナーから得られる、請求項1〜9のいずれか一項に記載の方法。
- 前記外分泌物質がドナー膵臓から得られる、請求項1〜10のいずれか一項に記載の方法。
- 前記島が前記膵組織から単離され、前記島が前記外分泌物質と組み合わされる、請求項2に記載の方法。
- 前記外分泌物質および前記島が単一のドナー膵臓から得られる、請求項1〜12のいずれか一項に記載の方法。
- 前記外分泌物質が合成である、または前記島の供給源ではないドナー膵臓から得られる、請求項1〜12のいずれか一項に記載の方法。
- 請求項1〜14のいずれか一項に記載の方法により調製される、または単離された膵島を単離された外分泌物質とエクスビボで組み合わせることにより得られる、パンクレアタイトを含む組成物。
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US20150132849A1 (en) * | 2011-09-28 | 2015-05-14 | Islet Sciences, Inc. | Ex vivo maturation of islet cells |
US20150361400A1 (en) * | 2013-01-25 | 2015-12-17 | Wake Forest University Health Sciences | Compositions and methods for maintaining and improving pancreatic islet cell function and stability |
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WO2023215876A1 (en) * | 2022-05-06 | 2023-11-09 | Tissue Genesis International Llc | Method for isolation of pancreatic islets |
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JP2010512148A (ja) * | 2006-12-07 | 2010-04-22 | ウィルソン ウォルフ マニュファクチャリング コーポレイション | 細胞培養に効果的な装置および方法 |
Family Cites Families (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5723282A (en) | 1991-07-08 | 1998-03-03 | The American National Red Cross | Method of preparing organs for vitrification |
US5599659A (en) | 1993-03-11 | 1997-02-04 | Breonics, Inc. | Preservation solution for ex vivo, warm preservation of tissues, explants,organs and vascular endothelial cells comprising retinal-derived fibroblast growth factor, cyclodextrin and chondroitin sulfate |
WO1994021116A1 (en) | 1993-03-16 | 1994-09-29 | Alliance Pharmaceutical Corp. | Preservation solution and method for warm organ preservation |
JP2813467B2 (ja) * | 1993-04-08 | 1998-10-22 | ヒューマン・セル・カルチャーズ・インコーポレーテッド | 細胞培養法および培地 |
US5405742A (en) | 1993-07-16 | 1995-04-11 | Cyromedical Sciences, Inc. | Solutions for tissue preservation and bloodless surgery and methods using same |
AU2595195A (en) | 1994-05-20 | 1995-12-18 | Vec Tec, Inc. | Method and apparatus monitoring viability of transplantable organs |
US5643712A (en) | 1994-05-20 | 1997-07-01 | Brasile; Lauren | Method for treating and rendering grafts nonthrombogenic and substantially nonimmunogenic using an extracellular matrix coating |
US5843024A (en) | 1996-05-17 | 1998-12-01 | Breonics, Inc. | Solution and process for resuscitation and preparation of ischemically damaged tissue |
US6815203B1 (en) * | 1999-06-23 | 2004-11-09 | Joslin Diabetes Center, Inc. | Methods of making pancreatic islet cells |
US6492103B1 (en) | 2000-01-31 | 2002-12-10 | Organ Recovery Systems, Inc. | System for organ and tissue preservation and hypothermic blood substitution |
AU2002365965A1 (en) * | 2001-11-19 | 2003-06-10 | University Of Miami | Improvement of viability and function of pancreatic islets |
BR0308753A (pt) * | 2002-03-22 | 2005-01-11 | Bayer Pharmaceuticals Corp | Método para produzir pseudo-ilhotas |
US7504201B2 (en) | 2004-04-05 | 2009-03-17 | Organ Recovery Systems | Method for perfusing an organ and for isolating cells from the organ |
US20060182722A1 (en) | 2005-02-11 | 2006-08-17 | Hering Bernhard J | Methods and materials for isolating isogenic islet cells |
US20080103606A1 (en) | 2006-10-30 | 2008-05-01 | Cory Berkland | Templated islet cells and small islet cell clusters for diabetes treatment |
WO2008063465A2 (en) * | 2006-11-13 | 2008-05-29 | Massachusetts Institute Of Technology | Encapsulated pancreatic islet cell products and methods of use thereof |
WO2010068728A2 (en) * | 2008-12-11 | 2010-06-17 | The Brigham And Women's Hospital, Inc. | Engineering functional tissue from cultured cells |
EP2546334A4 (en) * | 2010-03-12 | 2014-10-29 | Fundación Progreso Y Salud | PROCESS FOR IN VITRO PROLIFERATION OF CELLS FROM ENDODERMIC TISSUE |
US9834756B2 (en) * | 2010-07-16 | 2017-12-05 | Lifeline Scientific, Inc. | Methods for increasing isolation yields of cellular products |
-
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Non-Patent Citations (1)
Title |
---|
JPN6015039036; Cell Transplant. vol.19, no.5, 201001, pp.613-628 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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JPWO2016208459A1 (ja) * | 2015-06-22 | 2017-10-19 | 株式会社村田製作所 | 濾過装置、濾過方法、及び濾過フィルタ |
JP2019093387A (ja) * | 2015-06-22 | 2019-06-20 | 株式会社村田製作所 | 濾過装置、濾過方法、及び濾過フィルタ |
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