JP5921430B2 - 哺乳類細胞の心臓発生能を決定する方法 - Google Patents
哺乳類細胞の心臓発生能を決定する方法 Download PDFInfo
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Description
本出願は、2009年5月20日出願の国際出願番号PCT/US2009/044751について優先権の利益を主張する。先の出願の開示は、本出願の開示の一部とみなされる(また、本出願の開示に参考として援用される)。
骨髄の試料を、虚血性心疾患のために冠動脈バイパス術を受けている患者から収集した。管轄の倫理審査委員会により承認された方法により、患者からインフォームドコンセントを得た。
組換え型のTGFβ-1(2.5 ng/ml)、BMP-4 (5 ng/ml)、アクチビン−A(5 ng/ml)、FGF-2 (10 ng/ml)、α-トロンビン(1 U/ml)、IGF-1 (50 ng/ml)、カルジオトロフィン(1 ng/ml)、及びカルジオゲノールC(100 nM)を組み合わせて用い、これらを含有するカクテルで処理することによって、同様の結果が得られた。
組換え型のTGFβ-1(2.5 ng/ml)、BMP-4 (5 ng/ml)、アクチビン−A(5 ng/ml)、FGF-2 (10 ng/ml)、α-トロンビン (1 U/ml)、IGF-1 (50 ng/ml)、インターロイキン−6(100ng/ml)及びレチノイン酸(1μM)を組み合わせて用い、これらを含有するカクテルで処理することによって、同様の結果が得られた。
本発明は、その詳細な説明について記載されるが、上記説明は例示を目的としたものであり、添付の請求項の範囲により定められる本発明の範囲を限定するものではない。他の態様、効果、及び変更も、下記の請求項の範囲に含まれる。
Claims (13)
- 哺乳類細胞の心臓発生能を決定する方法であって、
a)前記細胞における、遺伝子の定量された発現量を収集する、ここで前記遺伝子は、Nkx2.5、Tbx5、MEF2C、GATA4、GATA6、Mesp1、及びFOG1(各遺伝子はその相同体であってもよい)である、工程と、
b)心臓発生能指数(CARPI)を決定する、ここで、心臓発生能指数(CARPI)は、前記細胞の前記遺伝子の定量された発現量の平均として計算される、工程と、
を含み、
前記遺伝子の発現量が、メッセンジャーRNA(mRNAs)、機能性RNA、又はこれらの組み合わせのRNA発現量で定量される
、方法。 - 前記機能性RNAが、マイクロRNAsである、請求項1に記載の方法。
- 前記細胞は、体細胞、生殖細胞、臍帯血細胞、心臓前駆細胞、胚性細胞、及びこれらのあらゆる組み合わせからなる群から選択される、請求項1又は2に記載の方法。
- 前記細胞は、遺伝子組み換えされている、請求項1〜3のいずれか一項に記載の方法。
- 前記細胞は、検出可能なサルコメアタンパク質を有しない、請求項1〜4のいずれか一項に記載の方法。
- 前記CARPIは、心臓発生性処置の前後の前記細胞について評価される、請求項1〜5のいずれか一項に記載の方法。
- 前記心臓発生性処置は、前記細胞を、細胞の心臓発生能を向上させることができる少なくとも2つの心臓発生性物質を含む組成物に接触させる工程を含む、請求項6に記載の方法。
- 前記細胞は、一種類の哺乳類に由来する、請求項1〜7のいずれか一項に記載の方法。
- 前記細胞は、哺乳類又は哺乳類群に由来し、該CARPIは、定量され、別の哺乳類又は哺乳類群由来の細胞のCARPIと比較される、請求項1〜7のいずれか一項に記載の方法。
- 前記CARPIは、下記式:
(式中、iは選択した遺伝子を表し、nは選択した遺伝子の全ての番号を表す)
を用いて、前記2つ以上の遺伝子の発現量の線形平均として計算される、請求項1〜9のいずれか一項に記載の方法。 - 前記CARPIは、処置の心臓発生能を定量的に評価するために測定される、請求項1〜10のいずれか一項に記載の方法。
- 前記CARPIは、心機能のパラメータと相関を有し、前記心機能のパラメータが心臓の駆出率である、請求項1〜11のいずれか一項に記載の方法。
- 処理装置と、該処理装置と組み合されて1つ又は複数の非神経性のネットワークプログラムをコード化するメモリーとを備えるコンピュータ装置であって、
該プログラムは、
a)哺乳類細胞における、遺伝子の定量された発現量を収集する、ここで前記遺伝子は、Nkx2.5、Tbx5、MEF2C、GATA4、GATA6、Mesp1、及びFOG1(各遺伝子はその相同体であってもよい)である、工程と、
b)心臓発生能指数(CARPI)を決定する、ここで、心臓発生能指数(CARPI)は、前記細胞の前記遺伝子の定量された発現量の平均として計算される、工程と、
c)前記CARPIを表示する工程と、
を含み、
前記遺伝子の発現量が、メッセンジャーRNA(mRNAs)、機能性RNA、又はこれらの組み合わせのRNA発現量で定量される、
方法を前記処理装置に実行させる、コンピュータ装置。
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USPCT/US2009/044751 | 2009-05-20 | ||
USPCT/US2009/044751 | 2009-05-20 | ||
PCT/US2010/035616 WO2010135555A1 (en) | 2009-05-20 | 2010-05-20 | Method for determining the cardio-generative potential of mammalian cells |
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US9765298B2 (en) | 2006-07-24 | 2017-09-19 | Mayo Foundation For Medical Education And Research | Methods and materials for providing cardiac cells |
WO2009145761A1 (en) | 2008-05-27 | 2009-12-03 | Mayo Foundation For Medical Education And Research | Methods and materials for using cells to treat heart tissue |
US9453205B2 (en) * | 2009-10-31 | 2016-09-27 | Genesis Technologies Limited | Methods for reprogramming cells and uses thereof |
KR101874463B1 (ko) * | 2009-10-31 | 2018-08-02 | 뉴 월드 레보러토리즈 인코포레이티드. | 세포의 재프로그램화 방법 및 이의 용도 |
US20140038291A1 (en) | 2009-10-31 | 2014-02-06 | New World Laboratories Inc. | Methods for reprogramming cells and uses thereof |
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US6324479B1 (en) * | 1998-05-08 | 2001-11-27 | Rosetta Impharmatics, Inc. | Methods of determining protein activity levels using gene expression profiles |
US6218122B1 (en) * | 1998-06-19 | 2001-04-17 | Rosetta Inpharmatics, Inc. | Methods of monitoring disease states and therapies using gene expression profiles |
US7547674B2 (en) * | 2001-06-06 | 2009-06-16 | New York Medical College | Methods and compositions for the repair and/or regeneration of damaged myocardium |
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