JP2013524787A - 糖尿病治療のためのテンプレート島細胞および小型島細胞クラスター - Google Patents
糖尿病治療のためのテンプレート島細胞および小型島細胞クラスター Download PDFInfo
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Abstract
【選択図】図1
Description
本出願は、米国特許出願第11/589,063号(2006年10月30日出願)の一部継続である。上記出願は、参照により本明細書中で援用される。
(連邦政府による資金提供を受けた研究または開発に関する陳述)
この明細書中で引用される特許出願、特許および出版物はすべて、これらの記載内容が参照により本明細書中で援用される。矛盾する場合は、本発明の開示(定義を含む)が優先される。
B. 多層として付着される島細胞
C. ディボット化微小鋳型上の島の産生
実施例1:島のサイズは生存度および移植成功に影響する
ラット島単離
収率
生存度
移植試験
実施例2:個々の島細胞または小型島細胞クラスターへの大型島の変換
a. 酵素消化
b. 金属ベースの断片化
c. 酵素消化および金属分散液の組合せ
実施例3:パッチ生体物質足場上での個々の島細胞および小型島クラスターの調製
足場物質のスクリーニング
島細胞パッチの調製
実施例4:生体物質足場上の島細胞の予測試験
実施例5:微小鋳型を用いた最適サイズ化細胞の調製
微小鋳型の開発
微小鋳型内の細胞再凝集
以下の参考文献は、それらが本明細書中に記述されるものを補足する例示的手法的またはその他の詳細を提供する範囲で、参照により本明細書中で援用される。
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, Dufrane D. In Vivo Selection of Biocompatible Alginates for Islet Encapsulation and Subcutaneous Transplantation, Tissue Eng Part A. Feb 11, [Epub ahead of print], (2010).
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Claims (25)
- 移植可能デバイスであって、
大表面を有する生体物質で構成される実質的に平坦な足場と、
前記生体物質の前記表面に付着されて、多層の島細胞を形成する個々の島細胞または小型島細胞クラスターであって、前記個々の島細胞または小型島細胞クラスターは、成体無傷島に由来し、前記小型島細胞クラスターが125μm未満の直径を有する、個々の島細胞または小型島細胞クラスターと、
を含む移植可能デバイス。 - 植込可能デバイスの生成方法であって、
膵臓から無傷島を得ることと、
前記無傷島から個々の島細胞または小型島細胞クラスターを取り出すことと、
多層の前記個々の島細胞および小型島細胞クラスターを実質的に平坦な植込可能生体物質足場の大表面に付着することと、
を包含する方法。 - 細胞を培養するための表面であって、
実質的に平坦な上面を含む支持体
を含み、
前記表面は、複数のディボットを含み、
各ディボットは、内部底面および内部側壁面を含み、
底面は、丸みを帯ており、
各ディボットは、直径100〜200μm(±20%)、深さ50〜150μm(±20%)であり、
細胞は個々のディボット中に分布される、表面。 - 請求項3記載の表面を含む細胞を培養するためのデバイスであって、
前記平坦表面を保持するための系をさらに含み、
前記系が底面および内部側壁面および内部容積を形成し、前記底および側壁が実質的に液密性であり、細胞および培地が内部容積中に分布される、デバイス。 - 前記細胞が島細胞である請求項3記載のデバイス。
- 前記細胞が非島細胞である請求項3記載のデバイス。
- 前記ディボットが直径100μm(±20%)および深さ60μm(±20%)である請求項3記載のデバイス。
- 前記平坦面がガラス製である請求項3記載の方法。
- 前記平坦面が生体適合性物質であり、ヒト患者における植込みに適している請求項3記載のデバイス。
- 滅菌性である請求項3記載のデバイス。
- 細胞の培養方法であって、
複数のディボットを含む表面に細胞を導入すること、
を含み、
各ディボットは、直径100〜200μm(±20%)および深さ50〜150μm(±20%)であり、前記デバイスを細胞培養条件に曝露する、方法。 - 前記細胞が島細胞であり、そしてデバイスおよび支持培養条件に曝露後に、前記細胞が小型島に再凝集する請求項11記載の方法。
- 前記細胞が約20〜150細胞/1ディボットの速度でディボット中に沈下する請求項11記載の方法。
- 前記細胞が島培養から分散され、化学物質または生物学的分子が結果的に生じる小型島中に組入れられる請求項11記載の方法。
- 前記細胞が非島細胞である請求項11記載の方法。
- 前記ディボットが直径100μm(±20%)および深さ60μm(±20%)である請求項11記載の方法。
- 導入化学物質に対する応答に関して複数の細胞試料を試験するための方法であって、
(a)複数のディボットを含むデバイス中で細胞を培養するステップであって、各ディボットは、直径100〜200μm(±20%)および深さ50〜150μm(±20%)である、細胞を培養するステップと、
(b)各ディボットに試験化学物質を導入するステップと、
(c)試験化学物質に対する細胞の応答を分析するステップと、
を包含する方法。 - 島の単離集団であって、
すべての島の少なくとも80%は、直径90μmより小さく、前記島がネイティブの大型島に比して低い拡散バリアを有し、
前記島は、ネイティブの小型および大型島に比して高い細胞生存度を有し、
島インスリン産生は、ネイティブ単離島より10倍以上大きいレベルにより特性化される、集団。 - 島の大多数が球形である請求項18記載の集団。
- 前記島が膵臓アルファ細胞、膵臓ベータ細胞および膵臓デルタ細胞である請求項18記載の集団。
- 前記島の少なくとも85%が生存可能である請求項18記載の集団。
- 全島の少なくとも80%が直径50μm未満である請求項18記載の集団。
- 前記島の少なくとも50%が直径37μm未満である請求項18記載の集団。
- 前記島ベータ細胞が、ネイティブ小型島またはネイティブ大型島から単離されるベータ細胞と比較してより多量にインスリンを産生する請求項18記載の集団。
- 前記島がネイティブ大型島に比して低拡散バリアを有する請求項18記載の集団。
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PCT/US2011/030775 WO2011126921A2 (en) | 2010-04-06 | 2011-03-31 | Templated islet cells and small islet cell clusters for diabetes treatment |
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US8895048B2 (en) | 2010-04-06 | 2014-11-25 | The University Of Kansas | Templated islet cells and small islet cell clusters for diabetes treatment |
JP2014521335A (ja) * | 2011-07-27 | 2014-08-28 | ユニバーシティ オブ カンザス | 糖尿病治療用膵島細胞および小膵島細胞クラスターテンプレート |
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WO2011126921A2 (en) | 2011-10-13 |
DK2555807T5 (da) | 2022-01-03 |
CA2795243C (en) | 2019-06-11 |
MX2012011644A (es) | 2013-02-11 |
US20140219997A1 (en) | 2014-08-07 |
BR112012025172B1 (pt) | 2018-08-28 |
EP2555807B9 (en) | 2021-12-08 |
DK2555807T3 (da) | 2019-11-11 |
CN102958543A (zh) | 2013-03-06 |
US8735154B2 (en) | 2014-05-27 |
US20100233239A1 (en) | 2010-09-16 |
JP5956422B2 (ja) | 2016-07-27 |
EP2555807A2 (en) | 2013-02-13 |
AU2011238540A1 (en) | 2012-11-29 |
AU2011238540B2 (en) | 2014-12-04 |
WO2011126921A3 (en) | 2012-01-19 |
BR112012025172A2 (pt) | 2017-06-27 |
CA2795243A1 (en) | 2011-10-13 |
EP2555807B1 (en) | 2019-07-31 |
CN102958543B (zh) | 2016-01-13 |
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