JP6076258B2 - 上皮細胞の不死化および使用法 - Google Patents
上皮細胞の不死化および使用法 Download PDFInfo
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Description
本出願は、2010年11月12日出願の米国仮出願第61/413,291号、および2011年4月13日出願の同第61/474,901号に対する優先権を主張するものであり、これら両方は、参照により組み込まれる。
本発明の開発中に行われた作業の一部は、米国国立衛生研究所(National Institutes of Health)の助成金交付番号R01CA106400およびR01−CA053371のもとで米国政府の資金を利用した。米国政府は、本発明においてある特定の権利を有する。
本発明は、非ケラチノサイト上皮細胞を培養する方法を対象とし、方法は、培養中に、フィーダー細胞、非ケラチノサイト上皮細胞、またはこれら両方におけるRhoキナーゼ(ROCK)の活性を阻害しながら、非ケラチノサイト上皮細胞をフィーダー細胞およびカルシウム含有培地の存在下で培養することを含む。本発明は、これらの不死化された非ケラチノサイト上皮細胞を使用する方法も対象とする。
配列番号2:アンチセンスプライマー
配列番号3:TaqManプローブ
配列番号4:プライマー
配列番号5:プライマー
配列番号6:プライマー
配列番号7:プライマー
配列番号8:センスプライマー
配列番号9:アンチセンスプライマー
配列番号10:TaqManプローブ
Claims (66)
- 非ケラチノサイト上皮細胞を連続的に培養する方法であって、
a)前記非ケラチノサイト上皮細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞およびカルシウム含有培地の存在下で培養すること、及び
b)培養中に、前記非ケラチノサイト上皮細胞におけるRhoキナーゼ(ROCK)の活性を阻害すること、
を含む、前記方法。 - 前記非ケラチノサイト上皮細胞は、初代細胞である、請求項1に記載の方法。
- 前記非ケラチノサイト上皮細胞は、初代細胞ではない、請求項1に記載の方法。
- 前記非ケラチノサイト上皮細胞は、腫瘍細胞である、請求項1に記載の方法。
- 前記非ケラチノサイト上皮細胞は、扁平上皮細胞、円柱細胞、腺腫細胞、および移行上皮細胞からなる群から選択される、請求項2、3、または4に記載の方法。
- 前記非ケラチノサイト上皮細胞は、前立腺細胞、乳房細胞、肝細胞、膵島細胞、肺上皮細胞、腎細胞、膀胱細胞、胃上皮細胞、大腸上皮細胞、小腸上皮細胞、尿道上皮細胞、精巣上皮細胞、卵巣上皮細胞、甲状腺細胞、副甲状腺細胞、副腎細胞、胸腺細胞、胆嚢細胞、および下垂体細胞からなる群から選択される、請求項5に記載の方法。
- 前記カルシウム含有培地は、血清または血清代替物を含む、請求項6に記載の方法。
- 前記フィーダー細胞は、増殖性線維芽細胞または非増殖性線維芽細胞である、請求項7に記載の方法。
- 前記非増殖性線維芽細胞は、マウス線維芽細胞またはヒト線維芽細胞である、請求項8に記載の方法。
- 前記ROCKは、Rhoキナーゼ阻害剤1(ROCK1)、Rhoキナーゼ阻害剤2(ROCK2)、またはこれら両方である、請求項8に記載の方法。
- ROCKの活性の阻害は、前記非ケラチノサイト上皮細胞を小分子ROCK阻害剤の存在下で培養することを含む、請求項10に記載の方法。
- 前記小分子ROCK阻害剤は、Y−27632、HA1100塩酸塩、HA1077、およびGSK429286からなる群から選択される、請求項11に記載の方法。
- ROCKの活性の阻害は、前記非ケラチノサイト上皮細胞をROCK1、ROCK2、またはこれら両方に特異的なRNA干渉(RNAi)分子の存在下で培養することを含む、請求項10に記載の方法。
- c)ROCKを阻害した後に、前記非ケラチノサイト上皮細胞を継代すること、及び
d)前記継代細胞をROCKが阻害されていない細胞培養環境下に設置すること、
をさらに含む、請求項1に記載の方法。 - 前記ROCKが阻害されていない環境は、三次元細胞培養環境である、請求項14に記載の方法。
- 前記ROCKが阻害されていない環境は、前記非ケラチノサイトの分化を通常誘導しない環境である、請求項14に記載の方法。
- 前記ROCKが阻害されていない環境は、前記非ケラチノサイトの分化を誘導する環境である、請求項14に記載の方法。
- (i)カルシウム含有培地及びフィーダー細胞、又は
(ii)フィーダー細胞で馴化されたカルシウム含有培地
の存在下で条件的に不死化された非ケラチノサイト上皮細胞集団であって、前記非ケラチノサイト上皮細胞においてRhoキナーゼ(ROCK)の活性が阻害されていることを特徴とする、細胞集団。 - 前記条件的に不死化された非ケラチノサイト上皮細胞は、正常な細胞、異常な細胞、又は罹患した細胞に由来する、請求項18に記載の細胞集団。
- 前記条件的に不死化された非ケラチノサイト上皮細胞は、腫瘍に由来する、請求項18に記載の細胞集団。
- 前記条件的に不死化された非ケラチノサイト上皮細胞は、扁平上皮細胞、円柱細胞、腺腫細胞、および移行上皮細胞からなる群から選択される細胞に由来する、請求項19または20に記載の細胞集団。
- 前記条件的に不死化された非ケラチノサイト上皮細胞は、前立腺細胞、乳房細胞、肝細胞、膵島細胞、肺上皮細胞、腎細胞、膀胱細胞、胃上皮細胞、大腸上皮細胞、小腸上皮細胞、尿道上皮細胞、精巣上皮細胞、卵巣上皮細胞、甲状腺細胞、副甲状腺細胞、副腎細胞、胸腺細胞、胆嚢細胞、および下垂体細胞からなる群から選択される細胞への分化を促進するその後の環境下に設置された後にそれらに分化することができる、請求項21に記載の細胞集団。
- 非ケラチノサイト上皮細胞の増殖を刺激する方法であって、
a)前記非ケラチノサイト上皮細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞およびカルシウム含有培地の存在下で培養すること、及び
b)培養中に、前記非ケラチノサイト上皮細胞におけるRhoキナーゼ(ROCK)の活性を阻害すること、を含み、
それによって、前記Rhoキナーゼの活性を阻害しながら前記非ケラチノサイト上皮細胞を培養することが、非ケラチノサイト上皮細胞の増殖を刺激する、
前記方法。 - 前記非ケラチノサイト上皮細胞は、初代細胞である、請求項23に記載の方法。
- 前記非ケラチノサイト上皮細胞は、初代細胞ではない、請求項23に記載の方法。
- 前記非ケラチノサイト上皮細胞は、腫瘍細胞である、請求項23に記載の方法。
- 前記非ケラチノサイト上皮細胞は、扁平上皮細胞、円柱細胞、腺腫細胞、および移行上皮細胞からなる群から選択される請求項24、25、または26に記載の方法。
- 前記非ケラチノサイト上皮細胞は、前立腺細胞、乳房細胞、肝細胞、膵島細胞、肺上皮細胞、腎細胞、膀胱細胞、胃上皮細胞、大腸上皮細胞、小腸上皮細胞、尿道上皮細胞、精巣上皮細胞、卵巣上皮細胞、甲状腺細胞、副甲状腺細胞、副腎細胞、胸腺細胞、胆嚢細胞、および下垂体細胞からなる群から選択される、請求項27に記載の方法。
- 前記カルシウム含有培地は、血清、血清代替物、又は血清代用物を含む、請求項28に記載の方法。
- 前記フィーダー細胞は、非増殖性線維芽細胞である、請求項29に記載の方法。
- 前記非増殖性線維芽細胞は、マウス線維芽細胞またはヒト線維芽細胞である、請求項30に記載の方法。
- 前記ROCKは、Rhoキナーゼ阻害剤1(ROCK1)、Rhoキナーゼ阻害剤2(ROCK2)、またはこれら両方である、請求項30に記載の方法。
- ROCKの活性の阻害は、前記非ケラチノサイト上皮細胞を小分子ROCK阻害剤の存在下で培養することを含む、請求項32に記載の方法。
- 前記小分子ROCK阻害剤は、Y−27632、HA1100塩酸塩、HA1077、およびGSK429286からなる群から選択される、請求項33に記載の方法。
- ROCKの活性の阻害は、前記非ケラチノサイト上皮細胞をROCK1、ROCK2、またはこれら両方に特異的なRNA干渉(RNAi)分子の存在下で培養することを含む、請求項32に記載の方法。
- 前記腫瘍細胞は、対象の血液循環から単離された、請求項26に記載の方法。
- 前記腫瘍細胞は、対象由来の組織から単離された、請求項26に記載の方法。
- 異常な非ケラチノサイト上皮(NKE)細胞の存在を特徴とする状態の治療を必要とする対象の候補治療を同定する方法であって、
a)前記対象から得られた前記異常なNKE細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞及びカルシウム含有培地、並びに少なくとも1つのRhoキナーゼ(ROCK)阻害剤の存在下で培養して、生体外で異常なNKE細胞集団を産生すること、
b)生体外で、前記異常なNKE細胞の少なくとも一部の応答プロファイルを決定すること、及び
c)前記決定された応答プロファイルに基づいて、前記対象の候補治療を同定すること、を含む、前記方法。 - 前記応答プロファイルは、生体外で、前記異常なNKE細胞から抽出されるDNAの少なくとも一部の配列を同定することによって少なくとも部分的に決定される、請求項38に記載の方法。
- 前記応答プロファイルは、生体外で、前記異常なNKE細胞中に産生される少なくとも1つのmRNAを同定することによって少なくとも部分的に決定される、請求項38に記載の方法。
- 前記応答プロファイルは、生体外で、前記異常なNKE細胞中に産生されない少なくとも1つのmRNAを同定することによって少なくとも部分的に決定される、請求項38に記載の方法。
- 前記応答プロファイルは、生体外で、前記異常なNKE細胞が発現する1つ以上のタンパク質を同定することによって少なくとも部分的に決定される、請求項38に記載の方法。
- 前記応答プロファイルは、生体外で、前記異常なNKE細胞が発現しない1つ以上のタンパク質を同定することによって少なくとも部分的に決定される、請求項38に記載の方法。
- 前記応答プロファイルは、生体外で、前記異常なNKE細胞を化学療法剤に供し、かつ、生体外で、前記異常なNKE細胞に対する前記化学療法剤の治療指数を決定することによって少なくとも部分的に決定される、請求項38に記載の方法。
- 前記対象由来の前記異常なNKE細胞は、前記対象の血液循環から得られる、請求項38に記載の方法。
- 対象における異常な非ケラチノサイト上皮(NKE)細胞を同定する方法であって、
a)前記対象から単離された少なくとも1つの異常な候補NKE細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞及びカルシウム含有培地、並びに少なくとも1つのRhoキナーゼ(ROCK)阻害剤の存在下で培養して、生体外で異常な候補NKE細胞集団を産生すること、
b)生体外で、前記異常な候補NKE細胞集団の少なくとも一部の組織起源プロファイルを決定すること、及び
c)前記異常な候補NKE細胞の少なくとも1つの特性を、前記異常な候補NKE細胞の前記決定された組織起源プロファイルの組織と同一の組織から得られる正常なNKE細胞の同一の特性と比較して、前記異常な候補NKE細胞と前記正常なNKE細胞との間に差異があるかを決定すること、を含み、
差異は、前記異常な候補NKE細胞が正常なNKE細胞と比較して異常であることを示す、
前記方法。 - 前記異常な候補NKE細胞は、前記対象の血液循環から得られる、請求項46に記載の方法。
- 前記組織起源プロファイルは、生体外で、前記異常な候補NKE細胞中に産生される少なくとも1つのmRNAを同定することによって少なくとも部分的に決定される、請求項46に記載の方法。
- 前記組織起源プロファイルは、生体外で、前記異常な候補NKE細胞中に産生されない少なくとも1つのmRNAを同定することによって少なくとも部分的に決定される、請求項46に記載の方法。
- 前記組織起源プロファイルは、生体外で、前記異常な候補NKE細胞が発現する1つ以上のタンパク質を同定することによって少なくとも部分的に決定される、請求項46に記載の方法。
- 前記組織起源プロファイルは、生体外で、前記異常な候補NKE細胞が発現しない1つ以上のタンパク質を同定することによって少なくとも部分的に決定される、請求項46に記載の方法。
- 前記組織起源プロファイルは、組織学的に少なくとも部分的に決定される、請求項46に記載の方法。
- 非ケラチノサイト上皮(NKE)細胞集団を製造する方法であって、
a)対象から得られたNKE細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞及びカルシウム含有培地、並びに少なくとも1つのRhoキナーゼ(ROCK)阻害剤の存在下で培養して、生体外のNKE細胞集団を産生すること、及び
b)前記生体外のNKE細胞集団を回収すること、
を含む、前記方法。 - 前記NKE細胞は、前立腺細胞、乳房細胞、肝細胞、膵島細胞、肺上皮細胞、腎細胞、膀胱細胞、胃上皮細胞、大腸および小腸上皮細胞、尿道上皮細胞、精巣上皮細胞、卵巣上皮細胞、子宮頸部上皮細胞、甲状腺細胞、副甲状腺細胞、副腎細胞、胸腺細胞、胆嚢細胞、ならびに下垂体細胞からなる群から選択される、請求項53に記載の方法。
- 遺伝子改変された非ケラチノサイト上皮(NKE)細胞集団を製造する方法であって、a)対象から得られたNKE細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞及びカルシウム含有培地、並びに少なくとも1つのRhoキナーゼ(ROCK)阻害剤の存在下で培養して、生体外のNKE細胞集団を産生すること、
b)生体外でNKE細胞集団の少なくとも一部を遺伝子改変すること、
c)遺伝子改変された前記NKE細胞を選択すること、
d)前記選択された遺伝子改変されたNKE細胞を(i)フィーダー細胞で馴化されたカルシウム含有培地、又は(ii)フィーダー細胞及びカルシウム含有培地、並びに少なくとも1つのRhoキナーゼ(ROCK)阻害剤の存在下で培養して、生体外の遺伝子改変されたNKE細胞集団を産生すること、及び
e)生体外で遺伝子改変された前記NKE細胞集団を回収すること、
を含む、前記方法。 - 前記対象から得られる前記NKE細胞は、肝細胞、肺上皮細胞、および膵島細胞からなる群から選択される、請求項55に記載の方法。
- 前記対象から得られる前記肝細胞は、第VIII因子のコーディング領域を含む導入遺伝子を挿入することによって遺伝子改変される、請求項55に記載の方法。
- 前記対象から得られる前記肺上皮細胞は、嚢胞性線維症膜コンダクタンス制御因子(CFTR)のコーディング領域を含む導入遺伝子を挿入することによって遺伝子改変される、請求項55に記載の方法。
- 前記対象由来の前記異常なNKE細胞は、前記対象由来の試料から得られる、請求項38に記載の方法。
- a)胚幹細胞でない、条件的に不死化された非ケラチノサイト上皮(NKE)細胞集団、b)カルシウム含有培地及びフィーダー細胞、又はフィーダー細胞で馴化されたカルシウム含有培地、並びに
c)カルシウム含有培地における少なくとも1つのRhoキナーゼ(Rock)阻害剤、を含む培養細胞組成物。 - 前記条件的に不死化されたNKE細胞は、初代扁平上皮細胞、初代円柱細胞、初代腺腫細胞、及び初代移行上皮細胞からなる群から選択される細胞に由来する、請求項60に記載の培養細胞組成物。
- 前記条件的に不死化されたNKE細胞は、分化を促進するその後の環境に設置された後に、不死化されていないNKE細胞が得られた組織型にのみ成熟することができ、前記不死化されていないNKE細胞が前立腺上皮細胞、乳房上皮細胞、肝細胞、膵島細胞、気道上皮細胞、腎臓上皮細胞、膀胱上皮細胞、胃上皮細胞、大腸上皮細胞、小腸上皮細胞、尿道上皮細胞、網膜上皮細胞、神経内分泌上皮細胞、精巣上皮細胞、卵巣上皮細胞、甲状腺上皮細胞、副甲状腺上皮細胞、副腎上皮細胞、胸腺上皮細胞、胆嚢上皮細胞、及び下垂体上皮細胞からなる群から選択される、請求項61に記載の培養細胞組成物。
- 前記カルシウム含有培地は、血清、血清代用物、又は血清代替物を含む、請求項60に記載の培養細胞組成物。
- 前記少なくとも1つのROCK阻害剤は、小分子Rhoキナーゼ1(ROCK1)阻害剤、小分子Rhoキナーゼ2(ROCK2)阻害剤、ROCK1を対象とするRNAi分子、ROCK2を対象とするRNAi分子、又はこれらの組合せである、請求項60に記載の培養細胞組成物。
- 前記小分子阻害剤は、Y−27632、HA1100塩酸塩、HA1077、及びGSK429286から選択される、請求項64に記載の培養細胞組成物。
- 対象への移植用である、請求項18〜22のいずれかに記載の細胞集団。
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WO2014082096A1 (en) | 2012-11-26 | 2014-05-30 | The Trustees Of Columbia University In The City Of New York | Method for culture of human and mouse prostate organoids and uses thereof |
JP6509745B2 (ja) * | 2013-01-25 | 2019-05-08 | エックスセル・バイオサイエンシズ・インコーポレイテッド | 標的細胞の選択的富化のための方法、組成物、キット、及びシステム |
WO2014172340A1 (en) * | 2013-04-15 | 2014-10-23 | Georgetown University | Immortalization of circulating tumor cells and methods of use |
WO2015192035A1 (en) * | 2014-06-13 | 2015-12-17 | Georgetown University | Compositions and methods for immortalization of epithelial cells |
JP2017522016A (ja) | 2014-06-27 | 2017-08-10 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | 培養哺乳動物輪部幹細胞、その産生方法及びその使用 |
KR102276424B1 (ko) | 2014-10-06 | 2021-07-12 | 삼성전자주식회사 | 로-키나제 저해제를 포함하는 세포의 노화를 감소시키기 위한 조성물 및 그의 용도 |
US10100285B2 (en) | 2015-04-03 | 2018-10-16 | Propagenix Inc. | Ex vivo proliferation of epithelial cells |
CA2981708A1 (en) | 2015-04-03 | 2016-10-06 | Chengkang ZHANG | Ex vivo proliferation of epithelial cells |
JP6869231B2 (ja) * | 2015-09-11 | 2021-05-12 | プロパジェニクス インコーポレイテッド | ex vivoでの上皮細胞の増殖 |
EP3426772A4 (en) | 2016-03-09 | 2019-08-28 | Beijing Percans Oncology Co. Ltd. | TUMOR CELL SUSPENSION CULTURES AND ASSOCIATED METHODS |
US20190151371A1 (en) * | 2016-06-09 | 2019-05-23 | Georgetown University | Process for continuous cell culture of islet cells |
CN107267446A (zh) * | 2017-07-27 | 2017-10-20 | 广州齐志生物工程设备有限公司 | 一种应用于条件性重编程技术的饲养细胞的处理方法 |
CN110093309B (zh) * | 2018-01-29 | 2021-07-02 | 中国科学院动物研究所 | 一种诱导成纤维细胞转分化为脂肪细胞的方法 |
EP3747991A4 (en) * | 2018-01-29 | 2022-01-19 | Institute Of Zoology, Chinese Academy Of Sciences | METHOD OF INDUCING CELLS |
CN110093310B (zh) * | 2018-01-29 | 2021-07-02 | 中国科学院动物研究所 | 一种将成纤维细胞转化为永生化细胞的方法及其应用 |
US20210147811A1 (en) * | 2018-04-09 | 2021-05-20 | Cedars-Sinai Medical Center | Methods for in vitro expansion of adult tissue stem cells |
KR20210005073A (ko) * | 2018-04-13 | 2021-01-13 | 상하이 라이드 바이오테크 컴퍼니 리미티드 | 조건부 리프로그래밍된 세포에서 동물 모델을 얻는 방법 및 항-종양 약물 스크리닝을 위한 동물 모델의 용도 |
CN112126627B (zh) * | 2020-09-30 | 2022-08-19 | 扬州大学 | 一种犬角膜上皮细胞永生化细胞系的构建方法和应用 |
WO2023085423A1 (ja) * | 2021-11-12 | 2023-05-19 | Ism株式会社 | 大腸上皮細胞の長期間培養 |
CN117018033A (zh) * | 2022-05-10 | 2023-11-10 | 上海赛立维生物科技有限公司 | 包含心肌前体样细胞的生物制剂及其制备方法和应用 |
Family Cites Families (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5665710A (en) | 1990-04-30 | 1997-09-09 | Georgetown University | Method of making liposomal oligodeoxynucleotide compositions |
JP3220180B2 (ja) | 1991-05-23 | 2001-10-22 | 三菱化学株式会社 | 薬剤含有タンパク質結合リポソーム |
NZ244306A (en) | 1991-09-30 | 1995-07-26 | Boehringer Ingelheim Int | Composition for introducing nucleic acid complexes into eucaryotic cells, complex containing nucleic acid and endosomolytic agent, peptide with endosomolytic domain and nucleic acid binding domain and preparation |
US5756122A (en) | 1995-06-07 | 1998-05-26 | Georgetown University | Liposomally encapsulated nucleic acids having high entrapment efficiencies, method of manufacturer and use thereof for transfection of targeted cells |
EP0843555B1 (en) | 1995-08-01 | 2003-08-27 | Isis Pharmaceuticals, Inc. | Liposomal oligonucleotide compositions |
US7217722B2 (en) | 2000-02-01 | 2007-05-15 | Kirin Beer Kabushiki Kaisha | Nitrogen-containing compounds having kinase inhibitory activity and drugs containing the same |
JPWO2002100833A1 (ja) | 2001-06-12 | 2004-09-24 | 住友製薬株式会社 | Rhoキナーゼ阻害剤 |
WO2003059913A1 (en) | 2002-01-10 | 2003-07-24 | Bayer Healthcare Ag | Roh-kinase inhibitors |
JP4469179B2 (ja) | 2002-01-23 | 2010-05-26 | バイエル ファーマセチカル コーポレーション | Rhoキナーゼ阻害剤としてのピリミジン誘導体 |
CA2473910C (en) | 2002-01-23 | 2011-03-15 | Bayer Pharmaceuticals Corporation | Pyrimidine derivatives as rho-kinase inhibitors |
TW200306819A (en) | 2002-01-25 | 2003-12-01 | Vertex Pharma | Indazole compounds useful as protein kinase inhibitors |
GB0206860D0 (en) | 2002-03-22 | 2002-05-01 | Glaxo Group Ltd | Compounds |
US7645878B2 (en) | 2002-03-22 | 2010-01-12 | Bayer Healthcare Llc | Process for preparing quinazoline Rho-kinase inhibitors and intermediates thereof |
EP1500643A4 (en) | 2002-04-03 | 2007-03-28 | Dainippon Sumitomo Pharma Co | BENZAMIDE DERIVATIVES |
ES2273047T3 (es) | 2002-10-28 | 2007-05-01 | Bayer Healthcare Ag | Fenilaminopirimidinas sustituidas con heteroariloxi como inhibidores de rho-cinasa. |
ES2374272T3 (es) | 2003-06-19 | 2012-02-15 | Glaxosmithkline Llc | Derivados de 5-(acilamino)indazol como inhibidores de quinasas. |
CA2530389A1 (en) | 2003-07-02 | 2005-01-13 | Biofocus Discovery Limited | Pyrazine and pyridine derivatives as rho kinase inhibitors |
US20100016329A1 (en) * | 2008-07-21 | 2010-01-21 | Kaohsiung Medical University | Xanthine-based cyclic gmp-enhancing rho-kinase inhibitor inhibits physiological activities of lung epithelial cell line |
ES2743204T3 (es) | 2008-11-04 | 2020-02-18 | Viacyte Inc | Composiciones de suspensión de agregados de células madre y métodos para su diferenciación |
US8008075B2 (en) * | 2008-11-04 | 2011-08-30 | Viacyte, Inc. | Stem cell aggregate suspension compositions and methods of differentiation thereof |
MX2011005288A (es) | 2008-11-20 | 2011-06-01 | Centocor Ortho Biotech Inc | Celulas madre pluripotentes en microportadores. |
WO2010065907A2 (en) * | 2008-12-05 | 2010-06-10 | The United States Of America As Represented By The Secretary, Department Of Health And Human Services | Use of a rock inhibitor to sustain primary human keratinocytes in a proliferative state |
EP3904505A1 (en) * | 2009-04-22 | 2021-11-03 | Viacyte, Inc. | Cell compositions derived from dedifferentiated reprogrammed cells |
US20130071927A1 (en) * | 2010-05-05 | 2013-03-21 | Sydney Ivf Limited | Media and methods for cell culture |
WO2012065067A2 (en) * | 2010-11-12 | 2012-05-18 | Georgetown University | Immortalization of epithelial cells and methods of use |
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