JP5819825B2 - ヒト胚性幹細胞の分化 - Google Patents
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Description
本発明は、米国特許公開番号第61/226,936号(2009年7月20日出願)に対する優先権を請求する。
本発明は、多能性幹細胞からのインスリン産生細胞への分化を促進させるための方法を提供する。具体的には、本発明は、動物への移植後にインスリンを産生することのできる細胞を産生するための方法を提供する。
a.多能性幹細胞を培養する工程、
b.多能性幹細胞を、胚体内胚葉系に特徴的なマーカーを発現している細胞へと分化させる工程、及び
c.胚体内胚葉系に特徴的なマーカーを発現している細胞を、FGF7を添加した第1培地で処理し、次いで細胞を、FGF7、BMP阻害能を持つ因子、TGF−β受容体アゴニスト、レチノイン酸、及びヘッジホッグ(hedgehog)シグナル経路阻害剤を添加した第2培地で培養することにより、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞へと分化させる工程。
幹細胞は、単一の細胞レベルにて自己複製し、分化して後代細胞を生成する、それら両方の能力で定義される未分化細胞であり、後代細胞としては、自己複製前駆細胞、非再生前駆細胞、及び最終分化細胞が挙げられる。幹細胞はまた、in vitroで複数の胚葉(内胚葉、中胚葉及び外胚葉)から様々な細胞系の機能的細胞へと分化する能力によって、また移植後に複数の胚葉の組織を生じ、胚盤胞への注入後、全部ではないとしてもほとんどの組織を提供する能力によっても、特徴付けられる。
多能性幹細胞の特徴付け
多能性幹細胞は、ステージ特異的胚抗原(SSEA)3及び4、並びにTra−1−60及びTra−1−81と呼ばれる抗体によって検出可能なマーカーのうちの1つ以上を発現している(Thomsonら、Science 282:1145,1998)。in vitroで多能性幹細胞を分化させると、SSEA−4、Tra−1−60、及びTra−1−81の発現が減少し(存在する場合)、SSEA−1の発現が上昇する。未分化の多能性幹細胞は通常アルカリホスファターゼ活性を有し、これは、細胞を4%パラホルムアルデヒドで固定した後、製造業者(Vector Laboratories(Burlingame Calif.))によって記載されるようにVectorRedを基質として現像することによって検出することができる。未分化の多能性幹細胞はまた、RT−PCRにより検出されるように、一般にOCT4及びTERTも発現する。
使用が可能な多能性幹細胞の種類としては、妊娠期間中の任意の時期(必ずしもではないが、通常は妊娠約10〜12週よりも前)に採取した前胚性組織(例えば胚盤胞など)、胚性組織、胎児組織などの、妊娠後に形成される組織に由来する多能性細胞の株化細胞系が挙げられる。非限定的な例は、例えばヒト胚幹細胞株H1、H7、及びH9(WiCell)などのヒト胚幹細胞又はヒト胚生殖細胞の確立株である。それらの細胞の最初の樹立又は安定化中に本開示の組成物を使用することも想定され、その場合、供給源となる細胞は、供給源となる組織から直接採取した一次多能性細胞であろう。フィーダー細胞の不在下で既に培養された多能性幹細胞集団から採取した細胞も好適である。例えば、BG01v(BresaGen、Athens、GA)などの変異ヒト胚性幹細胞株も好適である。
一実施形態では、多能性幹細胞は、典型的にはフィーダー細胞の層上で培養され、このフィーダー細胞は、多能性幹細胞を様々な方法で支持する。あるいは、多能性幹細胞を、フィーダー細胞を本質的に含まないにも関わらず、細胞を実質的に分化させることなく多能性幹細胞の増殖を支持するような培養システム中で培養する。フィーダー細胞不含培養における多能性幹細胞の、分化を伴わない増殖は、あらかじめ他の細胞種を培養することにより馴化培地を使用して支持される。あるいはフィーダー細胞不含培養における多能性幹細胞の分化を伴わない増殖は、合成培地を使用して支持される。
一実施形態では、本発明は、多能性幹細胞から、膵臓内胚葉系に特徴的なマーカーを発現している細胞を産生するための方法を提供し、この方法は以下の工程a、b及びcを包含する:
a.多能性幹細胞を培養する工程、
b.多能性幹細胞を、胚体内胚葉系に特徴的なマーカーを発現している細胞へと分化させる工程、及び
c.胚体内胚葉系に特徴的なマーカーを発現している細胞を、膵臓内胚葉系に特徴的なマーカーを発現している細胞へと分化させる工程。
胚体内胚葉系に特徴的なマーカーを発現している細胞の形成は、以下の特定のプロトコルの前後に、マーカーの存在に関して試験することにより決定することができる。多能性幹細胞は、一般にこのようなマーカーを発現しない。したがって、多能性細胞の分化は、細胞がそれらの発現を開始した際に検出される。
一実施形態では、胚体内胚葉系に特徴的なマーカーを発現している細胞を、FGF7を添加した第1培地で培養し、次いで細胞を、FGF7、BMP阻害能を持つ因子、TGFβ受容体アゴニスト、レチノイン酸、及びヘッジホッグ(hedgehog)シグナル経路阻害剤を添加した第2培地で培養することにより、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞へと分化させる。
一実施形態では、本発明の方法により産生される、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞は、膵内分泌系に特徴的なマーカーを発現している細胞へと更に分化し得る。
一態様では、本発明は、I型糖尿病に罹患しているかあるいはI型糖尿病を発症するリスクを有する患者を治療する方法を提供する。一実施形態では、本方法は、多能性幹細胞を培養することと、多能性幹細胞をin vitroでβ細胞系に分化させることと、β細胞系の細胞を患者に移植することと、を包含する。別の実施形態では、本方法は、多能性幹細胞を培養することと、多能性幹細胞を、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞へとin vitroで分化させることと、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない膵臓内胚葉系細胞を患者に移植することと、を包含する。
PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞へのヒト多能性幹細胞の分化
本実施例は、アクチビンAをノギン及びレチノイン酸と組み合わせて使用することで、NKX6.1発現の上方制御を促進できることを示す。簡潔に述べると、ヒト胚性幹細胞株H1細胞を、MATRIGEL(商標)(1:30希釈)をコートしたディッシュと、2%のBSA、100ng/mLのアクチビンA、20ng/mLのWNT−3a、8ng/mLのbFGFを添加したRPMI培地により1日培養した後で、2%のBSA、100ng/mLのアクチビンA、8ng/mLのbFGFを添加したRPMI培地で更に2日間処理し(ステージ1)、次いで
a.2%のBSA及び50ng/mLのFGF7を添加したDMEM/F12で3日間にわたって培養し(ステージ2)、次いで
b.1%のB27、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン、及び20ng/mLのアクチビンA若しくは50ng/mLのアクチビンAを添加したDMEM(高グルコース)で4日間にわたって培養した(ステージ3)。
ヒト胚性幹細胞株H1細胞を、MATRIGEL(1:30希釈)をコートしたプレート上で培養し、以下のプロトコルを使用して膵内分泌前駆細胞へと分化させた:
a.2%のBSA(カタログ# 152401,MP Biomedical,Ohio)、100ng/mLのアクチビンA(R&D Systems,MN)、20ng/mLのWNT−3a(カタログ# 1324−WN−002,R&D Systems,MN)及び8ng/mLのbFGF(カタログ# 100−18B,PeproTech,NJ)を添加したRPMI培地(カタログ#22400,Invitrogen,Ca)で1日培養した後に、2%のBSA、100ng/mLのアクチビンA、8ng/mLのbFGFを添加したRPMI培地で更に2日間にわたって処理し(ステージ1)、次いで
b.2%のBSA及び50ng/mLのFGF7を添加したDMEM/F12(カタログ#11330,Invitrogen,Ca)で3日間にわたって培養し(ステージ2)、次いで
c.処理1:1%のB27(Invitrogen,CA)、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)及び100ng/mLのノギンを添加したDMEM(高グルコース)で4日間にわたって処理するか(ステージ3)、又は
d.処理2:1%のB27(Invitrogen,CA)、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン及び20ng/mLのアクチビンAを添加したDMEM(高グルコース)で4日間にわたって処理するか(ステージ3)、又は
e.処理3:1%のB27(Invitrogen,CA)、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン及び1μMのALK5阻害剤IIを添加したDMEM(高グルコース)で4日間にわたって処理した(ステージ3)。
PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵内分泌前駆細胞への、膵臓内胚葉系に特徴的なマーカーを発現している細胞の分化
これまでの研究により、膵臓内胚葉系に特徴的なマーカーを発現している細胞は、更なる分化を受けた際に、インスリンを発現している細胞よりも、グルカゴンを発現している細胞を産生しやすいということが示されている。この示唆の一部は、膵臓内胚葉細胞でのNGN3発現に起因するものであり得る。本発明の方法は、NGN3を発現しない、膵臓内胚葉細胞集団を産生することから、これらの細胞集団はインスリンを発現している細胞へとより分化しやすいものと考えられる。しかしながら、NGN3発現は、膵内分泌細胞又は膵内分泌前駆細胞(例えばグルカゴン又はインスリンを発現している細胞を形成することのできる細胞)の形成に必要とされる。したがって、膵内分泌前駆細胞の最終的な運命を誘導するにあたり、NGN3の一時的な制御が重要である。
a.2%のBSA及び50ng/mLのFGF7を添加したDMEM/F12で3日間にわたって培養し(ステージ2)、次いで
b.1%のB27、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン及び20ng/mLのアクチビンAを添加したDMEM(高グルコース)で4日間にわたって処理し(ステージ3)、次いで
c.1%のB27、100ng/mLのノギン及び1μMのALK5阻害剤IIを添加したDMEM(高グルコース)で3日間にわたって培養するか(ステージ4)、又は
d.1%のB27のみを添加したDMEM(高グルコース)で3日間にわたって培養した(ステージ4)。
膵内分泌細胞への、PDX1とNKX6.1を共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞の分化
本実施例は、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉系に特徴的なマーカーを発現している細胞が、膵内分泌前駆細胞から、膵内分泌細胞へと更に分化するための能力について試験することを目的に設計された。
a.2%のBSA及び50ng/mLのFGF7を添加したDMEM/F12で3日間にわたって培養し(ステージ2)、次いで
b.1%のB27、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン及び20ng/mLのアクチビンAを添加したDMEM(高グルコース)で4日間にわたって処理するか(ステージ3)、又は
c.1%のB27、50ng/mLのFGF7、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン及び20ng/mLのアクチビンAを添加したDMEM(高グルコース)で4日間にわたって処理し(ステージ3)、次いで
d.1%のB27、100ng/mLのノギン及び1μMのALK5阻害剤IIを添加したDMEM(高グルコース)で3日間にわたって培養し(ステージ4)、
e.1%のB27、100ng/mLのノギン、1μMのALK5阻害剤II及び20ng/mLのベータセルリンを添加したDMEM(高グルコース)で5〜7日間にわたって培養した(ステージ5)。
STZ誘導型糖尿病の重症複合免疫不全(SCID)−beige(Bg)マウスへの、本発明の細胞の移植
ヒト胚性幹細胞株H1細胞を、MATRIGEL(登録商標)をコートしたディッシュ(1:30希釈)と、0.2%のFBS、100ng/mLのアクチビンA、20ng/mLのWNT−3aを添加したRPMI培地で1日培養し、次いで0.5%のFBS及び100ng/mLのアクチビンAを添加したRPMI培地により更に2日間にわたって処理し(ステージ1)、次いで
a.2%のFBS及び50ng/mLのFGF7を添加したDMEM/F12で3日間にわたって処理し(ステージ2)、次いで
b.1%のB27、0.25μMのシクロパミン−KAAD、2μMのレチノイン酸(RA)、100ng/mLのノギン、50ng/mLのFGF7及び20ng/mLのアクチビンAを添加したDMEM(高グルコース)で4日間にわたって処理し(ステージ3)、次いで
c.1%のB27、100ng/mLのノギン及び1μMのALK5阻害剤IIを添加したDMEM(高グルコース)で4日間にわたって処理した(ステージ4)。
Claims (1)
- PDX−1及びNKX−6.1は共発現するがCDX−2及びNGN−3は発現しない膵臓内胚葉細胞へと、多能性幹細胞集団を分化させる方法であって、以下の工程a、b及びcを包含する方法:
a.前記多能性幹細胞を培養する工程、
b.前記多能性幹細胞を、胚体内胚葉細胞へと分化させる工程、
c.前記胚体内胚葉細胞を、FGF7を添加した第1培地で処理し、次いでこうして得られる細胞を、FGF7、ノギン、レチノイン酸、シクロパミン−KAAD並びにアクチビンB、TGFβ−I、TGFβ−II、GFD−8及びGDF−IIからなる群より選ばれるTGF−β受容体アゴニストを添加した第2培地で培養することにより、PDX1及びNKX6.1は共発現するが、CDX2及びNGN3は発現しない、膵臓内胚葉細胞へと分化させる工程及び
d.PDX1及びNKX6.1は共発現するがCDX2及びNGN3は発現しない細胞を選別し、PDX1及びNKX6.1は共発現するがCDX2及びNGN3は発現しない膵臓内胚葉細胞を取得する工程。
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Families Citing this family (29)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009154606A1 (en) * | 2008-06-03 | 2009-12-23 | Cythera, Inc. | Growth factors for production of definitive endoderm |
US20100272695A1 (en) * | 2009-04-22 | 2010-10-28 | Alan Agulnick | Cell compositions derived from dedifferentiated reprogrammed cells |
SG10201501503VA (en) | 2010-03-01 | 2015-04-29 | Janssen Biotech Inc | Methods for purifying cells derived from pluripotent stem cells |
US8835400B2 (en) | 2010-10-08 | 2014-09-16 | Mina Therapeutics Limited | RNA molecules that upregulate insulin production |
CN108220224A (zh) * | 2011-06-21 | 2018-06-29 | 诺沃—诺迪斯克有限公司 | 自多潜能干细胞有效诱导定形内胚层 |
JP6441080B2 (ja) | 2011-12-22 | 2018-12-19 | ヤンセン バイオテツク,インコーポレーテツド | 単一ホルモンのインスリン陽性細胞へのヒト胚性幹細胞の分化 |
EP2859091B1 (en) * | 2012-06-08 | 2018-08-29 | Janssen Biotech, Inc. | Differentiation of human embryonic stem cells into pancreatic endocrine cells |
JP6470687B2 (ja) | 2012-09-03 | 2019-02-13 | ノヴォ ノルディスク アー/エス | 小分子を用いた多能性幹細胞からの膵臓内胚葉の作製 |
MX2015008619A (es) | 2012-12-31 | 2016-01-12 | Janssen Biotech Inc | Suspension y agrupamiento de celulas humanas pluripotentes para la diferenciacion a celulas endocrinas pancreaticas. |
WO2014105543A1 (en) | 2012-12-31 | 2014-07-03 | Janssen Biotech, Inc. | Culturing of human embryonic stem cells at the air-liquid interface for differentiation into pancreatic endocrine cells |
US8859286B2 (en) | 2013-03-14 | 2014-10-14 | Viacyte, Inc. | In vitro differentiation of pluripotent stem cells to pancreatic endoderm cells (PEC) and endocrine cells |
DE202014011287U1 (de) | 2013-06-11 | 2019-02-06 | The President And Fellows Of Harvard College | SC-ß Zellen und Zusammensetzungen zur Erzeugung der Zellen |
KR102252561B1 (ko) | 2013-11-22 | 2021-05-20 | 미나 테라퓨틱스 리미티드 | C/ebp 알파 짧은 활성화 rna 조성물 및 사용 방법 |
CN107614678B (zh) | 2014-12-18 | 2021-04-30 | 哈佛学院校长同事会 | 干细胞来源的β细胞的产生方法及其使用方法 |
WO2016100898A1 (en) | 2014-12-18 | 2016-06-23 | President And Fellows Of Harvard College | Serum-free in vitro directed differentiation protocol for generating stem cell-derived b cells and uses thereof |
EP3234110B1 (en) | 2014-12-18 | 2024-02-28 | President and Fellows of Harvard College | METHODS FOR GENERATING STEM CELL-DERIVED ß CELLS AND USES THEREOF |
US10767164B2 (en) | 2017-03-30 | 2020-09-08 | The Research Foundation For The State University Of New York | Microenvironments for self-assembly of islet organoids from stem cells differentiation |
US10391156B2 (en) | 2017-07-12 | 2019-08-27 | Viacyte, Inc. | University donor cells and related methods |
AU2018370029A1 (en) | 2017-11-15 | 2020-07-02 | Vertex Pharmaceuticals Incorporated | Islet cell manufacturing compositions and methods of use |
WO2020033879A1 (en) | 2018-08-10 | 2020-02-13 | Semma Therapeutics, Inc. | Stem cell derived islet differentiation |
US10724052B2 (en) | 2018-09-07 | 2020-07-28 | Crispr Therapeutics Ag | Universal donor cells |
WO2020068984A1 (en) * | 2018-09-25 | 2020-04-02 | The Regents Of The University Of California | Production and enrichment of pancreatic endocrine progenitor cells |
WO2020243665A1 (en) | 2019-05-31 | 2020-12-03 | W. L. Gore & Associates, Inc. | A biocompatible membrane composite |
EP3976236A1 (en) | 2019-05-31 | 2022-04-06 | W.L. Gore & Associates Inc. | A biocompatible membrane composite |
EP3976237A1 (en) | 2019-05-31 | 2022-04-06 | W.L. Gore & Associates Inc. | Cell encapsulation devices with controlled oxygen diffusion distances |
EP3975926A1 (en) | 2019-05-31 | 2022-04-06 | W.L. Gore & Associates, Inc. | A biocompatible membrane composite |
CA3150235A1 (en) | 2019-09-05 | 2021-03-11 | Alireza Rezania | UNIVERSAL DONOR CELLS |
CN114364791A (zh) | 2019-09-05 | 2022-04-15 | 克里斯珀医疗股份公司 | 通用供体细胞 |
EP4271796A1 (en) | 2020-12-31 | 2023-11-08 | CRISPR Therapeutics AG | Universal donor cells |
Family Cites Families (179)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3209652A (en) * | 1961-03-30 | 1965-10-05 | Burgsmueller Karl | Thread whirling method |
AT326803B (de) * | 1968-08-26 | 1975-12-29 | Binder Fa G | Maschenware sowie verfahren zur herstellung derselben |
US3935067A (en) * | 1974-11-22 | 1976-01-27 | Wyo-Ben Products, Inc. | Inorganic support for culture media |
US4499802A (en) * | 1982-09-29 | 1985-02-19 | Container Graphics Corporation | Rotary cutting die with scrap ejection |
US4537773A (en) * | 1983-12-05 | 1985-08-27 | E. I. Du Pont De Nemours And Company | α-Aminoboronic acid derivatives |
US4557264A (en) * | 1984-04-09 | 1985-12-10 | Ethicon Inc. | Surgical filament from polypropylene blended with polyethylene |
US5215893A (en) * | 1985-10-03 | 1993-06-01 | Genentech, Inc. | Nucleic acid encoding the ba chain prodomains of inhibin and method for synthesizing polypeptides using such nucleic acid |
US5089396A (en) * | 1985-10-03 | 1992-02-18 | Genentech, Inc. | Nucleic acid encoding β chain prodomains of inhibin and method for synthesizing polypeptides using such nucleic acid |
US4737578A (en) * | 1986-02-10 | 1988-04-12 | The Salk Institute For Biological Studies | Human inhibin |
US5863531A (en) | 1986-04-18 | 1999-01-26 | Advanced Tissue Sciences, Inc. | In vitro preparation of tubular tissue structures by stromal cell culture on a three-dimensional framework |
US5759830A (en) | 1986-11-20 | 1998-06-02 | Massachusetts Institute Of Technology | Three-dimensional fibrous scaffold containing attached cells for producing vascularized tissue in vivo |
US5567612A (en) | 1986-11-20 | 1996-10-22 | Massachusetts Institute Of Technology | Genitourinary cell-matrix structure for implantation into a human and a method of making |
CA1340581C (en) | 1986-11-20 | 1999-06-08 | Joseph P. Vacanti | Chimeric neomorphogenesis of organs by controlled cellular implantation using artificial matrices |
EP0363125A3 (en) | 1988-10-03 | 1990-08-16 | Hana Biologics Inc. | Proliferated pancreatic endocrine cell product and process |
US5837539A (en) * | 1990-11-16 | 1998-11-17 | Osiris Therapeutics, Inc. | Monoclonal antibodies for human mesenchymal stem cells |
US5449383A (en) * | 1992-03-18 | 1995-09-12 | Chatelier; Ronald C. | Cell growth substrates |
GB9206861D0 (en) * | 1992-03-28 | 1992-05-13 | Univ Manchester | Wound healing and treatment of fibrotic disorders |
CA2114282A1 (en) * | 1993-01-28 | 1994-07-29 | Lothar Schilder | Multi-layered implant |
JP3525221B2 (ja) | 1993-02-17 | 2004-05-10 | 味の素株式会社 | 免疫抑制剤 |
US5523226A (en) * | 1993-05-14 | 1996-06-04 | Biotechnology Research And Development Corp. | Transgenic swine compositions and methods |
GB9310557D0 (en) * | 1993-05-21 | 1993-07-07 | Smithkline Beecham Plc | Novel process and apparatus |
TW257671B (ja) * | 1993-11-19 | 1995-09-21 | Ciba Geigy | |
US5834308A (en) * | 1994-04-28 | 1998-11-10 | University Of Florida Research Foundation, Inc. | In vitro growth of functional islets of Langerhans |
US6001647A (en) * | 1994-04-28 | 1999-12-14 | Ixion Biotechnology, Inc. | In vitro growth of functional islets of Langerhans and in vivo uses thereof |
US6703017B1 (en) * | 1994-04-28 | 2004-03-09 | Ixion Biotechnology, Inc. | Reversal of insulin-dependent diabetes by islet-producing stem cells, islet progenitor cells and islet-like structures |
US6083903A (en) * | 1994-10-28 | 2000-07-04 | Leukosite, Inc. | Boronic ester and acid compounds, synthesis and uses |
WO1996020728A1 (fr) | 1994-12-29 | 1996-07-11 | Chugai Seiyaku Kabushiki Kaisha | Potentialisateur d'agent antitumoral comprenant un antagoniste de l'interleukine 6 |
US5843780A (en) | 1995-01-20 | 1998-12-01 | Wisconsin Alumni Research Foundation | Primate embryonic stem cells |
US5718922A (en) * | 1995-05-31 | 1998-02-17 | Schepens Eye Research Institute, Inc. | Intravitreal microsphere drug delivery and method of preparation |
US5908782A (en) * | 1995-06-05 | 1999-06-01 | Osiris Therapeutics, Inc. | Chemically defined medium for human mesenchymal stem cells |
KR100568438B1 (ko) | 1997-04-24 | 2006-04-07 | 오르토-맥네일 파마슈티칼, 인코퍼레이티드 | 염증성 질환의 치료에 유용한 치환된 이미다졸, 이의 제조방법 및 이를 포함하는 약제학적 조성물 |
AU8476698A (en) * | 1997-07-03 | 1999-01-25 | Osiris Therapeutics, Inc. | Human mesenchymal stem cells from peripheral blood |
US6670127B2 (en) * | 1997-09-16 | 2003-12-30 | Egea Biosciences, Inc. | Method for assembly of a polynucleotide encoding a target polypeptide |
EP1538206B1 (en) * | 1997-09-16 | 2010-03-24 | Centocor, Inc. | Method for the complete chemical synthesis and assembly of genes and genomes |
AU1197699A (en) | 1997-10-23 | 1999-05-10 | Geron Corporation | Methods and materials for the growth of primate-derived primordial stem cells |
US6372779B1 (en) | 1997-12-29 | 2002-04-16 | Ortho Pharmaceutical Corporation | Anti-inflammatory compounds |
ATE316795T1 (de) * | 1998-03-18 | 2006-02-15 | Osiris Therapeutics Inc | Mesenchymale stammzellen für die prävention und behandlung von immunantworten bei transplantationen |
MY132496A (en) | 1998-05-11 | 2007-10-31 | Vertex Pharma | Inhibitors of p38 |
US6667176B1 (en) * | 2000-01-11 | 2003-12-23 | Geron Corporation | cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells |
US7410798B2 (en) | 2001-01-10 | 2008-08-12 | Geron Corporation | Culture system for rapid expansion of human embryonic stem cells |
US6413556B1 (en) | 1999-01-08 | 2002-07-02 | Sky High, Llc | Aqueous anti-apoptotic compositions |
CA2359159A1 (en) * | 1999-01-21 | 2000-07-27 | Vitro Diagnostics, Inc. | Immortalized cell lines and methods of making the same |
US6815203B1 (en) * | 1999-06-23 | 2004-11-09 | Joslin Diabetes Center, Inc. | Methods of making pancreatic islet cells |
US6306424B1 (en) | 1999-06-30 | 2001-10-23 | Ethicon, Inc. | Foam composite for the repair or regeneration of tissue |
US6333029B1 (en) * | 1999-06-30 | 2001-12-25 | Ethicon, Inc. | Porous tissue scaffoldings for the repair of regeneration of tissue |
US6685936B2 (en) * | 1999-10-12 | 2004-02-03 | Osiris Therapeutics, Inc. | Suppressor cells induced by culture with mesenchymal stem cells for treatment of immune responses in transplantation |
US6303424B1 (en) * | 1999-10-21 | 2001-10-16 | United Microelectronics Corp. | Method for fabricating a buried bit line in a DRAM cell |
US20030082155A1 (en) * | 1999-12-06 | 2003-05-01 | Habener Joel F. | Stem cells of the islets of langerhans and their use in treating diabetes mellitus |
US6753153B2 (en) * | 1999-12-13 | 2004-06-22 | The Scripps Research Institute | Markers for identification and isolation of pancreatic islet α and β progenitors |
US7439064B2 (en) * | 2000-03-09 | 2008-10-21 | Wicell Research Institute, Inc. | Cultivation of human embryonic stem cells in the absence of feeder cells or without conditioned medium |
US7005252B1 (en) | 2000-03-09 | 2006-02-28 | Wisconsin Alumni Research Foundation | Serum free cultivation of primate embryonic stem cells |
US6436704B1 (en) * | 2000-04-10 | 2002-08-20 | Raven Biotechnologies, Inc. | Human pancreatic epithelial progenitor cells and methods of isolation and use thereof |
US6458589B1 (en) | 2000-04-27 | 2002-10-01 | Geron Corporation | Hepatocyte lineage cells derived from pluripotent stem cells |
CN1449439A (zh) * | 2000-06-26 | 2003-10-15 | 株式会社雷诺再生医学研究所 | 细胞级分包括能分化为神经细胞的细胞 |
KR100850812B1 (ko) | 2000-10-23 | 2008-08-06 | 스미스클라인 비참 코포레이션 | 신규 화합물 |
DK1362047T3 (da) | 2000-12-08 | 2006-09-04 | Ortho Mcneil Pharm Inc | Indazolylsubstituerede pyrrolinforbindelser som kinaseinhibitorer |
JP2004526676A (ja) | 2000-12-08 | 2004-09-02 | オーソ−マクニール・フアーマシユーチカル・インコーポレーテツド | キナーゼ阻害剤として有用な大員複素環式化合物 |
US6599323B2 (en) | 2000-12-21 | 2003-07-29 | Ethicon, Inc. | Reinforced tissue implants and methods of manufacture and use |
JP2005503759A (ja) * | 2001-01-24 | 2005-02-10 | アメリカ合衆国 | 幹細胞の膵臓内分泌細胞への分化方法 |
DK1355910T3 (da) * | 2001-01-25 | 2011-06-27 | Us Of America Represented By The Secretary Dept Of Health And Human Services | Formulering af borsyreforbindelser |
US6656488B2 (en) | 2001-04-11 | 2003-12-02 | Ethicon Endo-Surgery, Inc. | Bioabsorbable bag containing bioabsorbable materials of different bioabsorption rates for tissue engineering |
EP1379626A2 (en) * | 2001-04-19 | 2004-01-14 | DeveloGen Aktiengesellschaft für entwicklungsbiologische Forschung | A method for differentiating stem cells into insulin-producing cells |
JP4296781B2 (ja) | 2001-04-24 | 2009-07-15 | 味の素株式会社 | 幹細胞及びその分離方法 |
CA2447015A1 (en) | 2001-05-15 | 2002-11-21 | Rappaport Family Institute For Research In The Medical Sciences | Insulin producing cells derived from human embryonic stem cells |
US6626950B2 (en) | 2001-06-28 | 2003-09-30 | Ethicon, Inc. | Composite scaffold with post anchor for the repair and regeneration of tissue |
KR100418195B1 (ko) | 2001-07-05 | 2004-02-11 | 주식회사 우리기술 | 전력케이블의 다중절연진단장치 및 그 방법 |
GB0117583D0 (en) | 2001-07-19 | 2001-09-12 | Astrazeneca Ab | Novel compounds |
WO2003014313A2 (en) * | 2001-08-06 | 2003-02-20 | Bresagen, Ltd. | Alternative compositions and methods for the culture of stem cells |
US6617152B2 (en) * | 2001-09-04 | 2003-09-09 | Corning Inc | Method for creating a cell growth surface on a polymeric substrate |
KR20040054699A (ko) * | 2001-10-02 | 2004-06-25 | 엥스띠뛰 끌레이톤 드 라 러쉐르쉬 | 제한된 발현의 렌티바이러스 벡터 및 이의 적용과 관련된방법 및 조성물 |
WO2003033697A1 (en) | 2001-10-18 | 2003-04-24 | Ixion Biotechnology, Inc. | Conversion of liver stem and progenitor cells to pancreatic functional cells |
HUP0500699A3 (en) * | 2001-11-09 | 2010-01-28 | Artecel Sciences | Endocrine pancreas differentiation of adipose tissue-derived stromal cells and uses thereof |
EP1442115B9 (en) | 2001-11-15 | 2009-12-16 | Children's Medical Center Corporation | Methods of isolation, expansion and differentiation of fetal stem cells from chorionic villus, amniotic fluid, and placenta and therapeutic uses thereof |
KR20120003961A (ko) * | 2001-12-07 | 2012-01-11 | 사이토리 테라퓨틱스, 인크. | 처리된 리포애스퍼레이트 세포로 환자를 치료하기 위한 시스템 및 방법 |
EP2264146A1 (en) * | 2001-12-07 | 2010-12-22 | Geron Corporation | Islet cells from human embryonic stem cells |
AU2002218893A1 (en) | 2001-12-21 | 2003-07-09 | Thromb-X Nv | Compositions for the in vitro derivation and culture of embryonic stem (es) cell lines with germline transmission capability |
US20030162290A1 (en) | 2002-01-25 | 2003-08-28 | Kazutomo Inoue | Method for inducing differentiation of embryonic stem cells into functioning cells |
JPWO2003087349A1 (ja) * | 2002-04-17 | 2005-08-18 | 大塚製薬株式会社 | 間葉系細胞から膵β細胞を形成する方法 |
US20040161419A1 (en) * | 2002-04-19 | 2004-08-19 | Strom Stephen C. | Placental stem cells and uses thereof |
DE60319364T2 (de) | 2002-05-08 | 2009-02-19 | Janssen Pharmaceutica N.V. | Substituierte pyrroline als kinase inhibitoren |
US20060003446A1 (en) * | 2002-05-17 | 2006-01-05 | Gordon Keller | Mesoderm and definitive endoderm cell populations |
JP2006512046A (ja) | 2002-05-28 | 2006-04-13 | ベクトン・ディキンソン・アンド・カンパニー | invitroにおけるヒト膵臓腺房細胞の増殖およびインスリン産生細胞への分化転換のための方法 |
BR0311821A (pt) | 2002-06-05 | 2005-04-05 | Janssen Pharmaceutica Nv | Derivados de bisindolil-maleimid como inibidores de cinase |
GB0212976D0 (en) | 2002-06-06 | 2002-07-17 | Tonejet Corp Pty Ltd | Ejection method and apparatus |
CN1171991C (zh) | 2002-07-08 | 2004-10-20 | 徐如祥 | 人神经干细胞的培养方法 |
US6877147B2 (en) * | 2002-07-22 | 2005-04-05 | Broadcom Corporation | Technique to assess timing delay by use of layout quality analyzer comparison |
US7838290B2 (en) * | 2002-07-25 | 2010-11-23 | The Scripps Research Institute | Hematopoietic stem cells and methods of treatment of neovascular eye diseases therewith |
EP1539930A4 (en) | 2002-07-29 | 2006-08-09 | Es Cell Int Pte Ltd | METHOD IN MULTIPLE STAGES OF DIFFERENTIATION OF POSITIVE INSULIN-SENSITIVE CELLS, GLUCOSE |
US20040063204A1 (en) | 2002-08-14 | 2004-04-01 | Lijun Yang | Bone marrow cell differentiation |
EP1539928A4 (en) | 2002-09-06 | 2006-09-06 | Amcyte Inc | POSIOTIVE PANCREATIC ENDOCRINE PROGENITOR CELLS CD56 IN ADULT HUMAN BEINGS |
US9969977B2 (en) * | 2002-09-20 | 2018-05-15 | Garnet Biotherapeutics | Cell populations which co-express CD49c and CD90 |
US20040062753A1 (en) * | 2002-09-27 | 2004-04-01 | Alireza Rezania | Composite scaffolds seeded with mammalian cells |
AU2003285172A1 (en) | 2002-11-08 | 2004-06-03 | The Johns Hopkins University | Human embryonic stem cell cultures, and compositions and methods for growing same |
US7144999B2 (en) | 2002-11-23 | 2006-12-05 | Isis Pharmaceuticals, Inc. | Modulation of hypoxia-inducible factor 1 alpha expression |
EP1567639A4 (en) | 2002-12-05 | 2005-12-21 | Technion Res & Dev Foundation | CULTURE OF HUMAN PANCREATIC ISLANDS AND USES THEREOF |
JP4613069B2 (ja) | 2002-12-16 | 2011-01-12 | テクニオン リサーチ アンド ディベロップメント ファウンデーション リミテッド | 支持細胞非含有、異種非含有のヒト胚性幹細胞の調製方法およびこれらを使用して調製された幹細胞培養物 |
US20070155661A1 (en) | 2003-02-14 | 2007-07-05 | The Board Of Trustees Of The Leland Standord Junior University | Methods and compositions for modulating the development of stem cells |
US20070154981A1 (en) * | 2003-02-14 | 2007-07-05 | The Board Of Trustees Of The Leland Stanford Junior University | Insulin-producing cells derived from stem cells |
CA2520861A1 (en) | 2003-03-27 | 2004-10-14 | Ixion Biotechnology, Inc. | Method for transdifferentiation of non-pancreatic stem cells to the pancreatic pathway |
WO2004090110A2 (en) | 2003-03-31 | 2004-10-21 | Bresagen Inc. | Compositions and methods for the control, differentiation and/or manipulation of pluripotent cells through a gamma-secretase signaling pathway |
US20090203141A1 (en) | 2003-05-15 | 2009-08-13 | Shi-Lung Lin | Generation of tumor-free embryonic stem-like pluripotent cells using inducible recombinant RNA agents |
EP1641914B1 (en) * | 2003-06-27 | 2016-07-20 | DePuy Synthes Products, Inc. | Postpartum cells derived from placental tissue, and methods of making and using the same |
IL161903A0 (en) | 2003-07-17 | 2005-11-20 | Gamida Cell Ltd | Ex vivo progenitor and stem cell expansion for usein the treatment of disease of endodermally- deri ved organs |
ITRM20030395A1 (it) | 2003-08-12 | 2005-02-13 | Istituto Naz Per Le Malattie Infettive Lazz | Terreno di coltura per il mantenimento, la proliferazione e il differenziamento di cellule di mammifero. |
US7569385B2 (en) * | 2003-08-14 | 2009-08-04 | The Regents Of The University Of California | Multipotent amniotic fetal stem cells |
US7157275B2 (en) * | 2003-08-15 | 2007-01-02 | Becton, Dickinson And Company | Peptides for enhanced cell attachment and growth |
AU2004269395A1 (en) | 2003-08-27 | 2005-03-10 | Stemcells California, Inc. | Enriched pancreatic stem cell and progenitor cell populations, and methods for identifying, isolating and enriching for these populations |
JP2007515433A (ja) * | 2003-12-17 | 2007-06-14 | アラーガン インコーポレイテッド | Cyp26aおよびcyp26bの選択的阻害剤を使用するレチノイド反応性障害の処置方法 |
US20060030042A1 (en) | 2003-12-19 | 2006-02-09 | Ali Brivanlou | Maintenance of embryonic stem cells by the GSK-3 inhibitor 6-bromoindirubin-3'-oxime |
US7625753B2 (en) | 2003-12-23 | 2009-12-01 | Cythera, Inc. | Expansion of definitive endoderm cells |
US20050266554A1 (en) | 2004-04-27 | 2005-12-01 | D Amour Kevin A | PDX1 expressing endoderm |
US7704738B2 (en) | 2003-12-23 | 2010-04-27 | Cythera, Inc. | Definitive endoderm |
TWI334443B (en) * | 2003-12-31 | 2010-12-11 | Ind Tech Res Inst | Method of single cell culture of undifferentiated human embryonic stem cells |
WO2005065354A2 (en) | 2003-12-31 | 2005-07-21 | The Burnham Institute | Defined media for pluripotent stem cell culture |
WO2005071066A1 (en) | 2004-01-23 | 2005-08-04 | Board Of Regents, The University Of Texas System | Methods and compositions for preparing pancreatic insulin secreting cells |
GB2441530B (en) | 2004-02-12 | 2009-09-23 | Univ Newcastle | Stem Cells |
AU2005221095A1 (en) * | 2004-03-09 | 2005-09-22 | John J. O'neil | Methods for generating insulin-producing cells |
CN1950498A (zh) | 2004-03-10 | 2007-04-18 | 加利福尼亚大学董事会 | 培养胚胎干细胞的组合物和方法 |
WO2005097977A2 (en) | 2004-04-01 | 2005-10-20 | Wisconsin Alumni Research Foundation | Differentiation of stem cells to endoderm and pancreatic lineage |
JP4926946B2 (ja) | 2004-04-27 | 2012-05-09 | ヴィアサイト,インコーポレイテッド | Pdx1発現性内胚葉 |
JP5687816B2 (ja) | 2004-07-09 | 2015-03-25 | ヴィアサイト,インコーポレイテッド | 胚体内胚葉を分化させるための因子を同定する方法 |
JP5102030B2 (ja) | 2004-08-13 | 2012-12-19 | ユニバーシティ・オブ・ジョージア・リサーチ・ファウンデイション・インコーポレイテッド | ヒト胚性幹細胞における自己再生および分化のための組成物および方法 |
US20080268533A1 (en) | 2004-08-25 | 2008-10-30 | University Of Georgia Research Foundation, Inc. | Methods and Compositions Utilizing Myc and Gsk3Beta to Manipulate the Pluripotency of Embryonic Stem Cells |
DE102004043256B4 (de) | 2004-09-07 | 2013-09-19 | Rheinische Friedrich-Wilhelms-Universität Bonn | Skalierbarer Prozess zur Kultivierung undifferenzierter Stammzellen in Suspension |
MX2007002389A (es) | 2004-09-08 | 2009-02-12 | Wisconsin Alumni Res Found | Cultivo de celulas progenitoras embrionarias humanas. |
ES2383813T3 (es) | 2004-09-08 | 2012-06-26 | Wisconsin Alumni Research Foundation | Método de cultivo y cultivo de células madre embrionarias |
AU2006210955A1 (en) | 2005-01-31 | 2006-08-10 | Es Cell International Pte Ltd. | Directed differentiation of embryonic stem cells and uses thereof |
ES2627419T3 (es) | 2005-03-04 | 2017-07-28 | Lifescan, Inc. | Células estromales adultas derivadas del páncreas |
GB0505970D0 (en) | 2005-03-23 | 2005-04-27 | Univ Edinburgh | Culture medium containing kinase inhibitor, and uses thereof |
EP1876893B1 (en) | 2005-04-15 | 2012-04-11 | Geron Corporation | Cancer treatment by combined inhibition of proteasome and telomerase activities |
EP1874367B1 (en) | 2005-04-26 | 2011-07-06 | Arhus Universitet | Biocompatible material for surgical implants and cell guiding tissue culture surfaces |
MX2007015610A (es) | 2005-06-10 | 2008-02-21 | Irm Llc | Compuestos que mantienen la fluripotencia de las celulas totipotentes embrionarias. |
WO2006138433A2 (en) | 2005-06-14 | 2006-12-28 | The Regents Of The University Of California | Induction of cell differentiation by class i bhlh polypeptides |
EP1931764A1 (en) | 2005-06-21 | 2008-06-18 | GE Healthcare Bio-Sciences AB | Method for cell culture |
NZ564179A (en) | 2005-06-30 | 2010-09-30 | Janssen Pharmaceutica Nv | Cyclic anilino - pyridinotriazines as GSK-3 inhibitors |
US20080194021A1 (en) | 2005-07-29 | 2008-08-14 | Mays Robert W | Use of a Gsk-3 Inhibitor to Maintain Potency of Culture Cells |
CA2616863A1 (en) | 2005-07-29 | 2007-02-01 | Australian Stem Cell Centre Limited | Compositions and methods for growth of pluripotent cells |
KR20080056181A (ko) | 2005-09-02 | 2008-06-20 | 에이전시 포 사이언스, 테크놀로지 앤드 리서치 | 전구세포주의 유도 방법 |
GB2444686B (en) | 2005-09-12 | 2010-08-25 | Es Cell Int Pte Ltd | Differentiation of pluripotent stem cells using p38 MAPK inhibitors or prostaglandins |
CN101310012B (zh) | 2005-10-14 | 2012-05-09 | 明尼苏达大学董事会 | 非胚胎干细胞分化成具有胰腺表型的细胞 |
US20070122905A1 (en) | 2005-10-27 | 2007-05-31 | D Amour Kevin A | PDX1-expressing dorsal and ventral foregut endoderm |
WO2007082963A1 (es) | 2006-01-18 | 2007-07-26 | Fundación Instituto Valenciano De Infertilidad | Líneas de células madre embrionarias humanas y métodos para usar las mismas |
EP1994141B1 (en) | 2006-02-23 | 2017-11-15 | ViaCyte, Inc. | Compositions and methods useful for culturing differentiable cells |
US7695965B2 (en) | 2006-03-02 | 2010-04-13 | Cythera, Inc. | Methods of producing pancreatic hormones |
WO2007103282A2 (en) * | 2006-03-02 | 2007-09-13 | Cythera, Inc. | Endocrine precursor cells, pancreatic hormone-expressing cells and methods of production |
CA2650812C (en) | 2006-04-28 | 2017-12-12 | Lifescan, Inc. | Differentiation of human embryonic stem cells |
US8741643B2 (en) | 2006-04-28 | 2014-06-03 | Lifescan, Inc. | Differentiation of pluripotent stem cells to definitive endoderm lineage |
US20070259423A1 (en) * | 2006-05-02 | 2007-11-08 | Jon Odorico | Method of differentiating stem cells into cells of the endoderm and pancreatic lineage |
WO2007139929A2 (en) | 2006-05-25 | 2007-12-06 | The Burnham Institute For Medical Research | Methods for culture and production of single cell populations of human embryonic stem cells |
US8415153B2 (en) | 2006-06-19 | 2013-04-09 | Geron Corporation | Differentiation and enrichment of islet-like cells from human pluripotent stem cells |
CN100494359C (zh) | 2006-06-23 | 2009-06-03 | 中日友好医院 | 神经干细胞三维立体培养体外扩增的方法 |
EP2046946B8 (en) | 2006-06-26 | 2017-01-25 | Lifescan, Inc. | Pluripotent stem cell culture |
US20080003676A1 (en) | 2006-06-26 | 2008-01-03 | Millipore Corporation | Growth of embryonic stem cells |
US8968994B2 (en) | 2006-07-06 | 2015-03-03 | Jeremy Micah Crook | Method for stem cell culture and cells derived therefrom |
AU2007277364B2 (en) | 2006-07-26 | 2010-08-12 | Viacyte, Inc. | Methods of producing pancreatic hormones |
JP2008099662A (ja) | 2006-09-22 | 2008-05-01 | Institute Of Physical & Chemical Research | 幹細胞の培養方法 |
US20080091234A1 (en) * | 2006-09-26 | 2008-04-17 | Kladakis Stephanie M | Method for modifying a medical implant surface for promoting tissue growth |
US20100323442A1 (en) | 2006-10-17 | 2010-12-23 | Emmanuel Edward Baetge | Modulation of the phosphatidylinositol-3-kinase pathway in the differentiation of human embryonic stem cells |
CA2666789C (en) | 2006-10-18 | 2016-11-22 | Yong Zhao | Embryonic-like stem cells derived from adult human peripheral blood and methods of use |
TW200836749A (en) | 2007-01-09 | 2008-09-16 | Vioquest Pharmaceuticals Inc | Compositions including triciribine and bortezomib and derivatives thereof and methods of use thereof |
CN103627671A (zh) | 2007-01-30 | 2014-03-12 | 佐治亚大学研究基金会 | 产生中内胚层细胞及多能游走细胞的方法与细胞群及用途 |
GB0703188D0 (en) | 2007-02-19 | 2007-03-28 | Roger Land Building | Large scale production of stem cells |
EP3957716A1 (en) * | 2007-07-18 | 2022-02-23 | Janssen Biotech, Inc. | Differentiation of human embryonic stem cells |
EP2185693B1 (en) | 2007-07-31 | 2019-07-03 | Lifescan, Inc. | Differentiation of human embryonic stem cells |
KR101544498B1 (ko) | 2007-08-24 | 2015-08-17 | 스티칭 허트 네덜란드 칸커 인스티튜트 | 종양성 질환의 치료를 위한 조성물 |
US9062290B2 (en) | 2007-11-27 | 2015-06-23 | Lifescan, Inc. | Differentiation of human embryonic stem cells |
SG154367A1 (en) | 2008-01-31 | 2009-08-28 | Es Cell Int Pte Ltd | Method of differentiating stem cells |
EP2250252A2 (en) | 2008-02-11 | 2010-11-17 | Cambridge Enterprise Limited | Improved reprogramming of mammalian cells, and the cells obtained |
MX2010009251A (es) | 2008-02-21 | 2010-11-25 | Centocor Ortho Biotech Inc | Metodos, placas de superficie modificada y composiciones para la fijacion, el cultivo y el desprendimiento celular. |
EP2479260B1 (en) | 2008-03-17 | 2016-01-06 | Agency For Science, Technology And Research | Microcarriers for stem cell culture |
DK2283117T3 (da) | 2008-04-21 | 2014-01-20 | Viacyte Inc | Fremgangsmåde til oprensning af pancreatiske endodermceller afledt fra humane embryoniske stamceller |
US20090298178A1 (en) | 2008-06-03 | 2009-12-03 | D Amour Kevin Allen | Growth factors for production of definitive endoderm |
DE102008032236A1 (de) | 2008-06-30 | 2010-04-01 | Eberhard-Karls-Universität Tübingen | Isolierung und/oder Identifizierung von Stammzellen mit adipozytärem, chondrozytärem und pankreatischem Differenzierungspotential |
ES2697798T3 (es) | 2008-06-30 | 2019-01-28 | Janssen Biotech Inc | Diferenciación de células madre pluripotentes |
US20100028307A1 (en) | 2008-07-31 | 2010-02-04 | O'neil John J | Pluripotent stem cell differentiation |
CA2742268C (en) | 2008-10-31 | 2020-02-18 | Centocor Ortho Biotech Inc. | Differentiation of human embryonic stem cells to the pancreatic endocrine lineage |
US8008075B2 (en) | 2008-11-04 | 2011-08-30 | Viacyte, Inc. | Stem cell aggregate suspension compositions and methods of differentiation thereof |
GB2485113B (en) | 2009-07-20 | 2016-12-28 | Janssen Biotech Inc | Differentiation of human embryonic stem cells into cells of the pancreatic endoderm lineage |
SG10201501513TA (en) | 2010-03-02 | 2015-04-29 | Univ Singapore | Culture additives to boost stem cell proliferation and differentiation response |
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CN102482640B (zh) | 2015-03-11 |
AU2010276440B2 (en) | 2014-07-03 |
KR101893021B1 (ko) | 2018-08-29 |
EP2456858B1 (en) | 2018-08-29 |
MX340952B (es) | 2016-07-29 |
GB2485112B (en) | 2014-02-26 |
MX2012000899A (es) | 2012-02-13 |
HK1170262A1 (en) | 2013-02-22 |
ES2693088T3 (es) | 2018-12-07 |
WO2011011302A3 (en) | 2011-05-05 |
JP2012533321A (ja) | 2012-12-27 |
BR112012001557A2 (pt) | 2016-03-08 |
CA2768644A1 (en) | 2011-01-27 |
RU2540021C2 (ru) | 2015-01-27 |
EP2456858A2 (en) | 2012-05-30 |
EP2456858A4 (en) | 2015-03-18 |
AR077767A1 (es) | 2011-09-21 |
PL2456858T3 (pl) | 2019-01-31 |
CN102482640A (zh) | 2012-05-30 |
WO2011011302A2 (en) | 2011-01-27 |
KR20170117231A (ko) | 2017-10-20 |
GB201202847D0 (en) | 2012-04-04 |
KR101785626B1 (ko) | 2017-10-16 |
US20110014702A1 (en) | 2011-01-20 |
RU2012105923A (ru) | 2013-08-27 |
KR20120034793A (ko) | 2012-04-12 |
ZA201201221B (en) | 2013-07-31 |
SG177483A1 (en) | 2012-02-28 |
US8785184B2 (en) | 2014-07-22 |
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