JP5558097B2 - 効率的な核初期化方法 - Google Patents
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Description
本発明の好ましい態様によれば、該miRNAが、(a)胚性幹細胞において体細胞よりも高度に発現しており、かつ(b)該miRNAの存在下において該miRNAの非存在下の場合よりも高い核初期化効率を与える性質を有するmiRNAである上記の方法が提供される。
さらに本発明により、上記の方法により得られた人工多能性幹細胞を分化誘導して得られる各種細胞を用いて、化合物、薬剤、毒物などの生理作用や毒性を評価する方法が提供される。
組換えベクターを体細胞に導入する方法も特に限定されず、当業者に公知の方法で行うことができる。一時的トランスフェクション、微細注射、リン酸カルシウム沈澱法、リポソーム媒介トランスフェクション、DEAEデキストラン媒介トランスフェクション、エレクトロポレーション、遺伝子銃による方法などを用いることができる。
(a)Octファミリー遺伝子及びSoxファミリー遺伝子からなる2種の遺伝子の組み合わせ
(b)Octファミリー遺伝子、Klfファミリー遺伝子、及びSoxファミリー遺伝子からなる3種の遺伝子の組み合わせ;
(c)Octファミリー遺伝子、Soxファミリー遺伝子、Linファミリー遺伝子、及びNanog遺伝子からなる4種の遺伝子の組み合わせ;
などを挙げることができるが、これらに限定されることはない。
好ましい組み合わせとして、例えば、
(e)Octファミリー遺伝子、Klfファミリー遺伝子、Soxファミリー遺伝子、及びTERT遺伝子からなる4種の遺伝子の組み合わせ;
(f)Octファミリー遺伝子、Klfファミリー遺伝子、Soxファミリー遺伝子、及びSV40 Large T antigen遺伝子からなる4種の遺伝子の組み合わせ;
(g)Octファミリー遺伝子、Klfファミリー遺伝子、Soxファミリー遺伝子、TERT遺伝子、及びSV40 Large T antigen遺伝子からなる5種の遺伝子の組み合わせを挙げることができる。
必要に応じて、上記の組み合わせからKlfファミリー遺伝子を除くことができる場合もある。
(1)Oct3/4及びSox2からなる2種の遺伝子の組み合わせ;
(2)Oct3/4、Klf4、及びSox2からなる3種の遺伝子の組み合わせ;
(3)Oct3/4、Sox2、Lin28、及びNanogからなる4種の遺伝子の組み合わせ;
(4)Oct3/4、Sox2、TERT、及びSV40 Large T antigenからなる4種の遺伝子の組み合わせ;
(5)Oct3/4、Klf4、Sox2、TERT、及びSV40 Large T antigenからなる5種の遺伝子の組み合わせ
などであるが、これらに限定されることはない。
例1:マウス胎児線維芽細胞の核初期化による人工多能性幹細胞の調製
マウス由来Oct3/4、Sox2、及びKlf4の3種の遺伝子、コントロール用のDsRed、及び18種類のmiRNAのそれぞれをコードするpMXs-ベースのレトロウイルスベクターをPLAT-E細胞にFugene6 (Roche)を用いてトランスフェクトした。翌日、Nanog遺伝子のプロモーター領域の下流にEGFPとpuromycinの耐性遺伝子をコードした配列をつないだトランスジェニックマウス由来の胎児線維芽細胞(Nanog GFP MEF、WO2007/69666)を6 well plateに1×105個ずつまいた。翌日、この細胞にOct3/4、Sox2、及びKlf4、並びに18種類のmiRNAから選ばれる各1種類のmiRNAを発現するレトロウイルスを3因子の発現ウイルスの混合液1 ml+miRNA発現ウイルス液1 mlの割合で感染させ核初期化により人工多能性幹細胞の調製を行った。
マウス由来Oct3/4、Sox2、及びKlf4の3種の遺伝子、DsRed(コントロール)、mmu-miR-295のそれぞれをコードするpMXs-ベースのレトロウイルスベクターをPLAT-E細胞にFugene6 (Roche)を用いてトランスフェクトした。翌日、Nanog遺伝子のプロモーター領域の下流にEGFPとpuromycinの耐性遺伝子をコードした配列をつないだトランスジェニックマウス由来の尻尾線維芽細胞(Nanog GFP tailtip fibroblasts)を6 well plateに1×105個ずつまいた。翌日、この細胞にOct3/4、Sox2、及びKlf4の3因子とDsRedあるいはmmu-miR-295を発現するレトロウイルスを(1:1:1:1)の割合で感染させ核初期化により人工多能性幹細胞の調製を行った。
ヒト由来Oct3/4、Sox2、及びKlf4の3種の遺伝子に加え、コントロール用のDsRedと23種類のmiRNA又はmiRNAクラスターをコードするpMXs-ベースのレトロウイルスベクターをPLAT-E細胞にFugene6 (Roche)を用いてトランスフェクトした。翌日、げっ歯類のみに感染するウイルスのレセプターであるSlc7a1を発現するようにしたヒト成人皮膚細胞(adult human dermal fibroblasts)を6cm dishに3×105個ずつまいた。翌日、この細胞にOct3/4、Sox2、及びKlf4の3種の遺伝子と各種miRNAを発現するレトロウイルスを1:1:1:1の割合で感染させ核初期化による人工多能性幹細胞の生成を試みた。
Claims (7)
- 人工多能性幹細胞の製造方法であって、哺乳類動物の体細胞を、miRNAの存在下で核初期化因子により核初期化し、該人工多能性幹細胞を生成する工程を含み、該miRNAが、hsa-miR-372、hsa-miR-373、hsa-miR-302b、hsa-miR-302c、hsa-miR-302a、hsa-miR-302d、hsa-miR-367、hsa-miR-520c、mmu-miR-291a、mmu-miR-294、及びmmu-miR-295からなる群から選ばれる1又は2以上のRNA(ここで、該RNAの記号はmiRBaseデータベースの登録名を示す。)に含まれる1又は2以上のmiRNAであり、該核初期化因子が、Oct3/4、Klfファミリー、及びSoxファミリーからなる3種の遺伝子またはその遺伝子産物を含み、該Klfファミリーが、Klf2またはKlf4であり、該Soxファミリーが、Sox1、Sox2、Sox3、Sox15またはSox17であることを特徴とする前記方法。
- 前記miRNAがmiRBaseデータベースの登録名hsa-miR-302b-367で特定されるmiRNAである、請求項1に記載の方法。
- 前記miRNAが配列表の配列番号1〜12から選択される1又は2以上のRNA配列に含まれる1又は2以上のmiRNAである、請求項1又は2に記載の方法。
- 前記体細胞に、前記miRNAの発現を可能にするベクターを導入する工程をさらに含む、請求項1〜3のいずれか1項に記載の方法。
- 前記体細胞に、前記核初期化因子の発現を可能にするベクターを導入する工程をさらに含む、請求項1〜4のいずれか1項に記載の方法。
- KlfファミリーがKlf4である、請求項1〜5のいずれか1項に記載の方法。
- SoxファミリーがSox2である、請求項1〜6のいずれか1項に記載の方法。
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Families Citing this family (99)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9453219B2 (en) * | 2003-05-15 | 2016-09-27 | Mello Biotech Taiwan Co., Ltd. | Cosmetic designs and products using intronic RNA |
US8278104B2 (en) | 2005-12-13 | 2012-10-02 | Kyoto University | Induced pluripotent stem cells produced with Oct3/4, Klf4 and Sox2 |
US8129187B2 (en) | 2005-12-13 | 2012-03-06 | Kyoto University | Somatic cell reprogramming by retroviral vectors encoding Oct3/4. Klf4, c-Myc and Sox2 |
US20090227032A1 (en) * | 2005-12-13 | 2009-09-10 | Kyoto University | Nuclear reprogramming factor and induced pluripotent stem cells |
US8048999B2 (en) * | 2005-12-13 | 2011-11-01 | Kyoto University | Nuclear reprogramming factor |
US9213999B2 (en) * | 2007-06-15 | 2015-12-15 | Kyoto University | Providing iPSCs to a customer |
JP2008307007A (ja) * | 2007-06-15 | 2008-12-25 | Bayer Schering Pharma Ag | 出生後のヒト組織由来未分化幹細胞から誘導したヒト多能性幹細胞 |
WO2009075119A1 (ja) | 2007-12-10 | 2009-06-18 | Kyoto University | 効率的な核初期化方法 |
EP2072618A1 (en) * | 2007-12-14 | 2009-06-24 | Johannes Gutenberg-Universität Mainz | Use of RNA for reprogramming somatic cells |
EP2229444B1 (en) * | 2008-01-16 | 2019-10-30 | Shi-Lung Lin | Generation of tumor-free embryonic stem-like pluripotent cells using inducible recombinant rna agents |
CN101855350B (zh) * | 2008-05-02 | 2014-12-31 | 国立大学法人京都大学 | 核重编程序方法 |
US8852940B2 (en) | 2009-04-01 | 2014-10-07 | The Regents Of The University Of California | Embryonic stem cell specific microRNAs promote induced pluripotency |
JP5376478B2 (ja) | 2009-08-07 | 2013-12-25 | 国立大学法人京都大学 | 効率的な人工多能性幹細胞の樹立方法 |
GB0915523D0 (en) | 2009-09-07 | 2009-10-07 | Genome Res Ltd | Cells and methods for obtaining them |
GB0919773D0 (en) * | 2009-11-12 | 2009-12-30 | Univ Nottingham | Induced pluripotent stem cell |
WO2011102444A1 (ja) * | 2010-02-18 | 2011-08-25 | 国立大学法人大阪大学 | 誘導多能性幹細胞の製造方法 |
US9517250B2 (en) | 2010-04-28 | 2016-12-13 | The J. David Gladstone Institutes | Methods for generating cardiomyocytes |
EP2580328A2 (en) * | 2010-06-11 | 2013-04-17 | Cellartis AB | Micrornas for the detection and isolaton of human embryonic stem cell-derived cardiac cell types |
US9476041B2 (en) | 2010-07-12 | 2016-10-25 | National University Corporation Tottori University | Method for producing novel hipsc by means of siRNA introduction |
US9404082B2 (en) | 2010-12-03 | 2016-08-02 | Kyoto University | Method for production of eosinophil from pluripotent stem cell |
US9228204B2 (en) | 2011-02-14 | 2016-01-05 | University Of Utah Research Foundation | Constructs for making induced pluripotent stem cells |
WO2012151309A1 (en) * | 2011-05-02 | 2012-11-08 | Sanford-Burnham Medical Research Institute | Methods for regulating induced pluripotent stem cell generation and compositions thereof |
AU2012288249B2 (en) | 2011-07-25 | 2017-06-22 | Kyoto University | Method for screening induced pluripotent stem cells |
WO2013033213A1 (en) | 2011-08-30 | 2013-03-07 | The J. David Gladstone Institutes | Methods for generating cardiomyocytes |
JP6162604B2 (ja) | 2011-10-21 | 2017-07-12 | 国立大学法人京都大学 | 層流による多能性維持単一分散細胞培養法 |
JP5850702B2 (ja) * | 2011-10-25 | 2016-02-03 | 国立大学法人岐阜大学 | 間葉系細胞の分化調節剤およびこれを用いた医薬、並びに間葉系細胞への分化調節作用を有する物質のスクリーニング方法 |
US20140329317A1 (en) | 2011-11-25 | 2014-11-06 | Kyoto University | Method for culturing pluripotent stem cell |
EP2788033B1 (en) | 2011-12-05 | 2017-05-31 | Factor Bioscience Inc. | Methods and products for transfecting cells |
US8497124B2 (en) | 2011-12-05 | 2013-07-30 | Factor Bioscience Inc. | Methods and products for reprogramming cells to a less differentiated state |
EP2790736B1 (en) | 2011-12-12 | 2018-01-31 | Oncoimmunin, Inc. | In vivo delivery of oligonucleotides |
AU2013264708B2 (en) | 2012-05-23 | 2019-03-07 | Kyoto University | Highly efficient method for establishing artificial pluripotent stem cell |
CA2876868A1 (en) * | 2012-06-13 | 2013-12-19 | Stemgent, Inc. | Methods of preparing pluripotent stem cells |
WO2014037574A1 (en) * | 2012-09-10 | 2014-03-13 | Sanofi | Methods for reprogramming a somatic cell |
JP2014082956A (ja) | 2012-10-19 | 2014-05-12 | Somar Corp | 細胞培養基材、およびそれを用いた細胞培養方法並びに多能性幹細胞の分化誘導方法 |
RU2019143431A (ru) | 2012-11-01 | 2020-04-28 | Фэктор Байосайенс Инк. | Способы и продукты для экспрессии белков в клетках |
US20150247125A1 (en) * | 2012-11-02 | 2015-09-03 | Lonza Walkersville | Micrornas and cellular reprogramming |
WO2014123242A1 (ja) | 2013-02-08 | 2014-08-14 | 国立大学法人京都大学 | 巨核球及び血小板の製造方法 |
US9738861B2 (en) | 2013-03-06 | 2017-08-22 | Kyoto University | Culture system for pluripotent stem cells and method for subculturing pluripotent stem cells |
JP6473077B2 (ja) | 2013-03-21 | 2019-02-20 | 国立大学法人京都大学 | 神経分化誘導用の多能性幹細胞 |
EP2980207B1 (en) | 2013-03-25 | 2018-12-05 | Foundation for Biomedical Research and Innovation at Kobe | Cell sorting method |
CA2909230C (en) | 2013-04-12 | 2021-06-15 | Kyoto University | Method for inducing alveolar epithelial progenitor cells |
WO2014185358A1 (ja) | 2013-05-14 | 2014-11-20 | 国立大学法人京都大学 | 効率的な心筋細胞の誘導方法 |
US10159766B2 (en) | 2013-05-31 | 2018-12-25 | Iheart Japan Corporation | Layered cell sheet incorporating hydrogel |
US11225642B2 (en) | 2013-06-11 | 2022-01-18 | Kyoto University | Method for producing renal progenitor cells |
CN104293785A (zh) * | 2013-06-28 | 2015-01-21 | 四川大学华西医院 | microRNA302s在制备抗肿瘤药物中的用途 |
US9796962B2 (en) | 2013-08-07 | 2017-10-24 | Kyoto University | Method for generating pancreatic hormone-producing cells |
CN105492598B (zh) | 2013-08-29 | 2019-12-03 | 三浦典正 | 与细胞的抗衰老相关的生物分子群 |
WO2015034012A1 (ja) | 2013-09-05 | 2015-03-12 | 国立大学法人京都大学 | 新規ドーパミン産生神経前駆細胞の誘導方法 |
EP3064577B1 (en) | 2013-11-01 | 2020-09-09 | Kyoto University | Novel chondrocyte induction method |
WO2015073625A2 (en) * | 2013-11-15 | 2015-05-21 | The Mclean Hospital Corporation | Synergistic genome-nonintegrating reprogramming by micrornas and transcription factors |
MX2016009771A (es) | 2014-01-31 | 2016-11-14 | Factor Bioscience Inc | Metodos y productos para la produccion y administracion de acido nucleico. |
JP7012432B2 (ja) | 2014-07-14 | 2022-01-28 | 中外製薬株式会社 | タンパク質のエピトープを同定するための方法 |
WO2016104717A1 (ja) | 2014-12-26 | 2016-06-30 | 国立大学法人京都大学 | 肝細胞誘導方法 |
EP3543339A1 (en) | 2015-02-13 | 2019-09-25 | Factor Bioscience Inc. | Nucleic acid products and methods of administration thereof |
EP3280803B1 (en) | 2015-04-06 | 2021-05-26 | The Board of Trustees of the Leland Stanford Junior University | Chemically modified guide rnas for crispr/cas-mediated gene regulation |
EP3081638A1 (en) | 2015-04-16 | 2016-10-19 | Kyoto University | Method for producing pseudo-islets |
WO2016190899A1 (en) * | 2015-05-23 | 2016-12-01 | Beem Alan M H | Pri-mirna libraries and methods for making and using pri-mirna libraries |
KR102312123B1 (ko) | 2016-04-22 | 2021-10-13 | 고쿠리츠 다이가쿠 호진 교토 다이가쿠 | 도파민 생산 신경전구세포의 제조방법 |
EP3455346A1 (en) | 2016-05-12 | 2019-03-20 | Erasmus University Medical Center Rotterdam | A method for culturing myogenic cells, cultures obtained therefrom, screening methods, and cell culture medium |
US10221395B2 (en) * | 2016-06-16 | 2019-03-05 | Cedars-Sinai Medical Center | Efficient method for reprogramming blood to induced pluripotent stem cells |
CN116115629A (zh) | 2016-08-17 | 2023-05-16 | 菲克特生物科学股份有限公司 | 核酸产品及其施用方法 |
EP4431607A2 (en) | 2016-09-09 | 2024-09-18 | The Board of Trustees of the Leland Stanford Junior University | High-throughput precision genome editing |
CN110114470A (zh) | 2016-12-27 | 2019-08-09 | 住友化学株式会社 | 诱导的多能性干细胞的评价方法及选择方法、以及诱导的多能性干细胞的制备方法 |
WO2018135646A1 (ja) | 2017-01-20 | 2018-07-26 | 国立大学法人京都大学 | CD8α+β+細胞傷害性T細胞の製造方法 |
WO2018139548A1 (ja) | 2017-01-26 | 2018-08-02 | 国立大学法人大阪大学 | 幹細胞の中胚葉系細胞への分化誘導用培地および中胚葉系細胞の製造方法 |
JP7171055B2 (ja) | 2017-03-14 | 2022-11-15 | 国立大学法人京都大学 | 多能性幹細胞からヘルパーt細胞を製造する方法 |
WO2018216743A1 (ja) | 2017-05-25 | 2018-11-29 | 国立大学法人京都大学 | 中間中胚葉細胞から腎前駆細胞への分化誘導方法、および多能性幹細胞から腎前駆細胞への分化誘導方法 |
KR102422175B1 (ko) | 2017-06-19 | 2022-07-18 | 고에키 자이단 호징 고베 이료 산교 도시 스이신 기코 | 만능 줄기 세포의 분화능의 예측 방법 및 이를 위한 시약 |
NL2019517B1 (en) | 2017-09-08 | 2019-03-19 | Univ Erasmus Med Ct Rotterdam | New therapy for Pompe disease |
EP3699267A4 (en) | 2017-10-17 | 2021-10-27 | Kyoto University | METHOD OF MANUFACTURING AN ARTIFICIAL NEUROMUSCULAR END PLATE FROM PLURIPOTENT STEM CELLS |
EP3822342A4 (en) | 2018-07-13 | 2022-08-03 | Kyoto University | PROCESS FOR PRODUCTION OF GAMMA DELTA T LYMPHOCYTES |
US20210299331A1 (en) | 2018-07-19 | 2021-09-30 | Kyoto University | Pluripotent stem cell-derived plate-shaped cartilage and method for producing the same |
WO2020022261A1 (ja) | 2018-07-23 | 2020-01-30 | 国立大学法人京都大学 | 新規腎前駆細胞マーカーおよびそれを利用した腎前駆細胞の濃縮方法 |
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US20220056413A1 (en) | 2018-12-21 | 2022-02-24 | Kyoto University | Lubricin-localized cartilage-like tissue, method for producing same and composition comprising same for treating articular cartilage damage |
EP3903884A4 (en) | 2018-12-26 | 2023-01-25 | Kirin Holdings Kabushiki Kaisha | MODIFIED TCR AND MANUFACTURING PROCESS THEREOF |
WO2020235319A1 (ja) | 2019-05-20 | 2020-11-26 | 味の素株式会社 | 軟骨又は骨の前駆細胞の拡大培養方法 |
US10501404B1 (en) | 2019-07-30 | 2019-12-10 | Factor Bioscience Inc. | Cationic lipids and transfection methods |
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MX2022010521A (es) | 2020-02-28 | 2022-09-19 | Takeda Pharmaceuticals Co | Metodo para producir linfocitos citoliticos naturales a partir de celulas madre pluripotentes. |
EP3922431A1 (en) | 2020-06-08 | 2021-12-15 | Erasmus University Medical Center Rotterdam | Method of manufacturing microdevices for lab-on-chip applications |
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KR20230030618A (ko) | 2020-07-01 | 2023-03-06 | 엘레바테바이오 테크놀로지스, 인코포레이티드 | 원형 rna를 사용하는 세포 리프로그래밍을 위한 조성물 및 방법 |
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US20230323289A1 (en) | 2020-07-20 | 2023-10-12 | Aichi Medical University | Composition for Pluripotent Cell Undifferentiated State Maintenance Culture, Medium for Pluripotent Cell Undifferentiated State Maintenance Culture, Method for Pluripotent Cell Undifferentiated State Maintenance Culture, and Method for Producing Pluripotent Cells |
WO2022039279A1 (ja) | 2020-08-18 | 2022-02-24 | 国立大学法人京都大学 | ヒト始原生殖細胞/ヒト始原生殖細胞様細胞の維持増幅方法 |
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JPWO2022230977A1 (ja) | 2021-04-30 | 2022-11-03 | ||
WO2022255489A1 (ja) | 2021-06-04 | 2022-12-08 | キリンホールディングス株式会社 | 細胞組成物、細胞組成物の製造方法及び細胞組成物を含む医薬組成物 |
JPWO2022259721A1 (ja) | 2021-06-10 | 2022-12-15 | ||
WO2022260718A1 (en) * | 2021-06-12 | 2022-12-15 | Lin Shi Lung | Novel replicase cycling reaction (rcr) |
AR126145A1 (es) | 2021-06-15 | 2023-09-13 | Takeda Pharmaceuticals Co | Método para producir linfocitos citolíticos naturales a partir de células madre pluripotentes |
CN113462638B (zh) * | 2021-06-30 | 2022-10-25 | 呈诺再生医学科技(珠海横琴新区)有限公司 | 一种高效无遗传修饰的iPSC诱导、产业化单克隆挑取平台及应用 |
US20240335480A1 (en) | 2021-07-15 | 2024-10-10 | Astellas Pharma Inc. | Vascular endothelial growth factor (vegf)-highly expressing pericyte-like cell |
WO2023286832A1 (ja) | 2021-07-15 | 2023-01-19 | アステラス製薬株式会社 | 血管内皮増殖因子(vegf)高発現ペリサイト様細胞の製造方法 |
US20240318142A1 (en) | 2021-08-11 | 2024-09-26 | Kyoto University | Method for producing renal interstitial progenitor cells, erythropoietin-producing cells, and method for producing renin-producing cells |
JP2024534945A (ja) | 2021-09-10 | 2024-09-26 | アジレント・テクノロジーズ・インク | 化学修飾を有するプライム編集のためのガイドrna |
EP4431605A1 (en) | 2021-11-11 | 2024-09-18 | Healios K.K. | Gene-modified pluripotent stem cell, immunocompetent cell derived therefrom, method for producing said cells, and use thereof |
JPWO2023153464A1 (ja) | 2022-02-09 | 2023-08-17 |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004024940A2 (en) * | 2002-09-16 | 2004-03-25 | University Of Southern California | Rna-mediated gene modulation |
WO2007069666A1 (ja) * | 2005-12-13 | 2007-06-21 | Kyoto University | 核初期化因子 |
WO2008118820A2 (en) * | 2007-03-23 | 2008-10-02 | Wisconsin Alumni Research Foundation | Somatic cell reprogramming |
WO2008124133A1 (en) * | 2007-04-07 | 2008-10-16 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
WO2009007852A2 (en) * | 2007-06-15 | 2009-01-15 | Izumi Bio, Inc | Multipotent/pluripotent cells and methods |
WO2009058413A1 (en) * | 2007-10-29 | 2009-05-07 | Shi-Lung Lin | Generation of human embryonic stem-like cells using intronic rna |
JP2010158171A (ja) * | 2008-12-08 | 2010-07-22 | Kyoto Univ | 効率的な核初期化方法 |
Family Cites Families (32)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003523166A (ja) | 1998-09-29 | 2003-08-05 | ガミダ セル リミテッド | 幹細胞および前駆細胞の増殖と分化を制御する方法 |
US6667176B1 (en) | 2000-01-11 | 2003-12-23 | Geron Corporation | cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells |
CA2349415A1 (en) | 1998-11-09 | 2000-05-18 | Monash University | Embryonic stem cells |
US6280718B1 (en) | 1999-11-08 | 2001-08-28 | Wisconsin Alumni Reasearch Foundation | Hematopoietic differentiation of human pluripotent embryonic stem cells |
US6458589B1 (en) | 2000-04-27 | 2002-10-01 | Geron Corporation | Hepatocyte lineage cells derived from pluripotent stem cells |
JP2002065261A (ja) | 2000-08-30 | 2002-03-05 | Mitsubishi Kasei Institute Of Life Sciences | 生殖細胞の取得方法 |
CA2430653A1 (en) | 2000-11-27 | 2002-08-08 | Yissum Research Development Company Of The Hebrew University In Jerusalem | Transfection of human embryonic stem cells |
JP2003009854A (ja) | 2001-04-09 | 2003-01-14 | Kyowa Hakko Kogyo Co Ltd | エンブリオイドボディ形成方法及びその用途 |
EP1403366B1 (en) | 2001-05-31 | 2015-05-27 | Shinya Yamanaka | Genes with es cell-specific expression |
US20030044976A1 (en) | 2001-08-27 | 2003-03-06 | Advanced Cell Technology | De-differentiation and re-differentiation of somatic cells and production of cells for cell therapies |
JP2004248505A (ja) | 2001-09-21 | 2004-09-09 | Norio Nakatsuji | 移植抗原の一部または全てを欠除したes細胞由来の未分化な体細胞融合細胞およびその製造 |
AU2002330465B2 (en) | 2001-09-21 | 2006-07-27 | Kyoto University | Screening method for a reprogramming agent, reprogramming agent screened by the method, method for using the reprogramming agent, method for differentiating an undifferentiated fusion cell, and production method of cells, tissues and organs. |
JP3736517B2 (ja) | 2002-11-13 | 2006-01-18 | 学校法人近畿大学 | 体細胞核初期化因子 |
AU2003901099A0 (en) | 2003-03-11 | 2003-03-27 | Es Cell International Pte Ltd. | Methods of inducing differentiation of stem cells |
US9567591B2 (en) * | 2003-05-15 | 2017-02-14 | Mello Biotechnology, Inc. | Generation of human embryonic stem-like cells using intronic RNA |
JP2005095027A (ja) | 2003-09-22 | 2005-04-14 | Reprocell Inc | 細胞の未分化状態マーカープロモーターおよびその利用 |
US7682828B2 (en) | 2003-11-26 | 2010-03-23 | Whitehead Institute For Biomedical Research | Methods for reprogramming somatic cells |
US7964401B2 (en) | 2004-02-19 | 2011-06-21 | Kyoto University | Screening method for somatic cell nuclear reprogramming substance affecting ECAT2 and ECAT3 |
WO2005090557A1 (ja) | 2004-03-23 | 2005-09-29 | Daiichi Asubio Pharma Co., Ltd. | 多能性幹細胞の増殖方法 |
JPWO2006035741A1 (ja) | 2004-09-29 | 2008-05-15 | 伸弥 山中 | Es細胞特異的発現遺伝子及びその利用 |
WO2007026255A2 (en) | 2005-06-22 | 2007-03-08 | Universitetet I Oslo | Dedifferentiated cells and methods of making and using dedifferentiated cells |
US8278104B2 (en) | 2005-12-13 | 2012-10-02 | Kyoto University | Induced pluripotent stem cells produced with Oct3/4, Klf4 and Sox2 |
US20090227032A1 (en) | 2005-12-13 | 2009-09-10 | Kyoto University | Nuclear reprogramming factor and induced pluripotent stem cells |
CN101389761A (zh) | 2006-02-27 | 2009-03-18 | 银怎株式会社 | 使用bmi-1使星形胶质细胞去分化成为神经干细胞 |
US20090252711A1 (en) | 2006-05-11 | 2009-10-08 | Andrew Craig Boquest | Stem Cells And Methods Of Making And Using Stem Cells |
WO2008030610A2 (en) | 2006-09-08 | 2008-03-13 | Michigan State University | Human transcriptome corresponding to human oocytes and use of said genes or the corresponding polypeptides to trans-differentiate somatic cells |
WO2008105630A1 (en) | 2007-02-27 | 2008-09-04 | Procell Therapeutics Inc. | Combined use of cell permeable nanog and oct4 for increasing self-renewal and suppressing differentiation of stem cells |
JP2010528622A (ja) | 2007-05-30 | 2010-08-26 | ザ ジェネラル ホスピタル コーポレイション | 体細胞から多能性細胞を生成する方法 |
WO2009032456A2 (en) | 2007-08-01 | 2009-03-12 | Primegen Biotech Llc | Non-viral delivery of transcription factors that reprogram human somatic cells into a stem cell-like state |
CA2660123C (en) | 2007-10-31 | 2017-05-09 | Kyoto University | Nuclear reprogramming method |
WO2009075119A1 (ja) | 2007-12-10 | 2009-06-18 | Kyoto University | 効率的な核初期化方法 |
US9683232B2 (en) | 2007-12-10 | 2017-06-20 | Kyoto University | Efficient method for nuclear reprogramming |
-
2008
- 2008-05-23 WO PCT/JP2008/059586 patent/WO2009075119A1/ja active Application Filing
- 2008-05-23 AU AU2008286249A patent/AU2008286249B2/en active Active
- 2008-05-23 KR KR1020097005179A patent/KR101532442B1/ko active IP Right Grant
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- 2008-11-25 US US12/292,717 patent/US20100075421A1/en not_active Abandoned
-
2011
- 2011-12-07 US US13/313,670 patent/US8791248B2/en active Active
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004024940A2 (en) * | 2002-09-16 | 2004-03-25 | University Of Southern California | Rna-mediated gene modulation |
WO2007069666A1 (ja) * | 2005-12-13 | 2007-06-21 | Kyoto University | 核初期化因子 |
WO2008118820A2 (en) * | 2007-03-23 | 2008-10-02 | Wisconsin Alumni Research Foundation | Somatic cell reprogramming |
WO2008124133A1 (en) * | 2007-04-07 | 2008-10-16 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
WO2009007852A2 (en) * | 2007-06-15 | 2009-01-15 | Izumi Bio, Inc | Multipotent/pluripotent cells and methods |
WO2009058413A1 (en) * | 2007-10-29 | 2009-05-07 | Shi-Lung Lin | Generation of human embryonic stem-like cells using intronic rna |
JP2010158171A (ja) * | 2008-12-08 | 2010-07-22 | Kyoto Univ | 効率的な核初期化方法 |
Non-Patent Citations (1)
Title |
---|
JPN6013035147; Nature Biotechnology Vol.27, No.5, 20090518, p.459-461 * |
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US8791248B2 (en) | 2014-07-29 |
US20130102768A1 (en) | 2013-04-25 |
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WO2009075119A1 (ja) | 2009-06-18 |
AU2008286249A1 (en) | 2009-06-25 |
US20100075421A1 (en) | 2010-03-25 |
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