JPWO2011125948A1 - Es細胞の製造方法 - Google Patents
Es細胞の製造方法 Download PDFInfo
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Abstract
Description
[1]副腎皮質刺激ホルモン、FGF受容体阻害剤、MEK活性化阻害剤及びGSK3阻害剤を含む培地中で、哺乳動物の内細胞塊を培養すること、及び培養物から哺乳動物ES細胞を単離することを含む、哺乳動物ES細胞の製造方法。
[2]哺乳動物がマウスである、[1]記載の製造方法。
[3]マウスがNOD/SCIDバックグラウンドマウスである、[2]記載の製造方法。
[4]培地が更にLIFを含む、[1]記載の製造方法。
[5]FGF受容体阻害剤がSU5402である、[1]記載の製造方法。
[6]MEK活性化阻害剤がPD184352である、[1]記載の製造方法。
[7]GSK3阻害剤がCHIR99021である、[1]記載の製造方法。
[8]多能性を維持したまま増殖可能な、単離されたNOD/SCIDバックグラウンドマウスES細胞。
[9]副腎皮質刺激ホルモン、FGF受容体阻害剤、MEK活性化阻害剤及びGSK3阻害剤を含む培地。
[10]更にLIFを含む、[9]記載の培地。
NOD/SCIDバックグラウンドマウスのES細胞を用いれば、インビトロでこのES細胞に様々な遺伝子操作を行い、容易に遺伝子改変NOD/SCIDバックグラウンドマウスを製造することが可能となる。従って、NOD/SCIDバックグラウンドマウスのES細胞は、ヒト化マウス製造の基盤として有用である。
(方法)
NOD/SCIDマウスを自然交配後、2日目に胚を卵管より採取した。8細胞胚を単離し、M16培地においてCO2インキュベーター(7%CO2、37℃)を用いて2日間培養し、胚盤胞期まで発生させた。胚盤胞期胚をマイトマイシンC処理したマウス胎児性線維芽細胞[MEF]を敷き詰めた4穴ディッシュに移し、以下の組成の培地(CCM+3i培地)中で培養した。胚がMEF上に付着し、内細胞塊(ICM)の成長が始まるまで培地の交換は行わなかった(約1週間程度)。
Glasgow minimal essential medium(GMEM,Sigma)
10%(v/v) FCS;
100μM 2−メルカプトエタノール(Nacalai tesuque);
1×非必須アミノ酸(Invitrogen);
1mM ピルビン酸ナトリウム(Invitrogen);
2000units/mL LIF(ESGRO,Invitrogen);
10μm ACTH;及び
3i(FGF受容体阻害剤 SU5402、2μM;MEK活性化の阻害剤 PD184352、0.8μM;及びGSK3阻害剤 CHIR99021、3μM)
28個の着床前胚の培養を行い、9個のES様細胞(雌5クローン、雄4クローン)を得た。このうち、雄由来2クローンについて、C57BL/6由来胚盤胞胚への微小注入をそれぞれ40回あるいは42回行い、それぞれ6個体(雌3、雄3)および9個体(雌3、雄6)のキメラマウスを得た(図1、2)。このうち、雄個体について生殖系列への伝達をチェックし、2個体における伝達を確認した(図3、4)。
CCM+3i培地の代わりに、以下の組成を有する培地(比較培地1)を用いて、NOD/SCID/コモンガンマKOマウス着床前胚21個を、実施例1と同一のプロトコールを用いて培養し、2クローンのES様細胞の作成に成功した。
Glasgow minimal essential medium(GMEM,Sigma)
10%(v/v) FCS;
100μM 2−メルカプトエタノール(Nacalai tesuque);
1×非必須アミノ酸(Invitrogen);
1mM ピルビン酸ナトリウム(Invitrogen);
10000units/mL LIF(ESGRO, Invitrogen);及び
10μm ACTH
(比較培地1は、3iを含まず、LIFの濃度が10000units/mLである点で、CCM+3i培地と異なる。)
実施例1と同一のプロトコールに従い、BALB/cAマウスの着床前胚からES細胞を樹立した。比較として、CCM+3i培地の代わりに、以下の組成を有する培地(比較培地2)を用いて、ES細胞の樹立を試みた。
Glasgow minimal essential medium(GMEM,Sigma)
10%(v/v) FCS;
100μM 2−メルカプトエタノール(Nacalai tesuque);
1×非必須アミノ酸(Invitrogen);
1mM ピルビン酸ナトリウム(Invitrogen);
2000units/mL LIF(ESGRO, Invitrogen);及び
3i(FGF受容体阻害剤 SU5402、2μM;MEK活性化の阻害剤 PD184352、0.8μM;及びGSK3阻害剤 CHIR99021、3μM)
(比較培地2は、ACTHを含まない点で、CCM+3i培地と異なる。)
(方法)
NOD/SCID由来ES細胞を、下記の培地(CCM+3i培地)中で2X107までフィーダー細胞であるMEF上で増幅した後、電気穿孔法を用いた通常のプロトコールにより、線状化したヒト・エリスロポイエチン遺伝子ノックインベクター(図11)を細胞内に導入した。
Glasgow minimal essential medium(GMEM,Sigma)
10%(v/v) FCS;
100μM 2−メルカプトエタノール(Nacalai tesuque);
1×非必須アミノ酸(Invitrogen);
1mM ピルビン酸ナトリウム(Invitrogen);
2000units/mL LIF(ESGRO,Invitrogen);
10μm ACTH;及び
3i(FGF受容体阻害剤 SU5402、2μM;MEK活性化の阻害剤 PD184352、0.8μM;及びGSK3阻害剤 CHIR99021、3μM)
NOD/SCIDバックグラウンドマウスのES細胞を用いれば、インビトロでこのES細胞に様々な遺伝子操作を行い、容易に遺伝子改変NOD/SCIDバックグラウンドマウスを製造することが可能となる。従って、NOD/SCIDバックグラウンドマウスのES細胞は、ヒト化マウス製造の基盤として有用である。
Claims (10)
- 副腎皮質刺激ホルモン、FGF受容体阻害剤、MEK活性化阻害剤及びGSK3阻害剤を含む培地中で、哺乳動物の内細胞塊を培養すること、及び培養物から哺乳動物ES細胞を単離することを含む、哺乳動物ES細胞の製造方法。
- 哺乳動物がマウスである、請求項1記載の製造方法。
- マウスがNOD/SCIDバックグラウンドマウスである、請求項2記載の製造方法。
- 培地が更にLIFを含む、請求項1記載の製造方法。
- FGF受容体阻害剤がSU5402である、請求項1記載の製造方法。
- MEK活性化阻害剤がPD184352である、請求項1記載の製造方法。
- GSK3阻害剤がCHIR99021である、請求項1記載の製造方法。
- 多能性を維持したまま増殖可能な、単離されたNOD/SCIDバックグラウンドマウスES細胞。
- 副腎皮質刺激ホルモン、FGF受容体阻害剤、MEK活性化阻害剤及びGSK3阻害剤を含む培地。
- 更にLIFを含む、請求項9記載の培地。
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JP2009545302A (ja) * | 2006-08-01 | 2009-12-24 | ザ・ユニバーシティ・コート・オブ・ザ・ユニバーシティ・オブ・エディンバラ | ラットおよび他種由来の多能性細胞 |
Family Cites Families (7)
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---|---|---|---|---|
US4415546A (en) | 1981-05-12 | 1983-11-15 | Janakiraman Ramachandran | Biologically active analogs of ACTH and radioimmuno assay therefor |
PT87133B (pt) | 1987-04-02 | 1992-07-31 | Amrad Corp Ltd | Metodo de purificacao do factor inibidor da leucemia (lif) e de composicoes farmaceuticas contendo polipeptidos com actividade do lif |
DE69028514T2 (de) | 1989-01-10 | 1997-02-20 | Amrad Corp Ltd | Leukämiehemmender faktor aus vieharten und seine verwendung zur verbesserung der implantation und der entwicklung von embryonalen zellen |
JPH08154681A (ja) | 1994-12-01 | 1996-06-18 | Ito Ham Kk | ウシlif 遺伝子 |
JPH0994040A (ja) | 1995-09-29 | 1997-04-08 | Kanagawa Kagaku Gijutsu Akad | 免疫不全マウス及びその作出方法 |
CA2402459A1 (en) | 2000-12-01 | 2002-06-06 | Central Institute For Experimental Animals | Method of producing a mouse suitable for engraftment, differentiation and proliferation of heterologous cells, mouse produced by this method and use of the mouse |
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Non-Patent Citations (7)
Title |
---|
JPN6015015986; Nature 453, 2008, 519-523 * |
JPN6015015988; Cell 135, 2008, 1287-1298 * |
JPN6015015989; Biol. Reprod. 81, 2009, 1147-1153 * |
JPN6015015990; Cloning and Stem Cells 10, 2008, 503-512 * |
JPN6015015991; Genesis 48, 20100216, 317-327 * |
JPN6015015992; Genes to Cells 9, 2004, 471-477 * |
JPN6015015993; Stem cells 27, 2009, 383-389 * |
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EP2554659A1 (en) | 2013-02-06 |
WO2011125948A1 (ja) | 2011-10-13 |
EP2554659A4 (en) | 2013-09-25 |
US20130059375A1 (en) | 2013-03-07 |
JP2016063841A (ja) | 2016-04-28 |
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