JP6678107B2 - 膵前駆細胞の増殖方法 - Google Patents
膵前駆細胞の増殖方法 Download PDFInfo
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Description
[1]膵前駆細胞を以下の工程(1)に付すことを特徴とする、膵前駆細胞の増殖方法(本明細書中、本発明の増殖方法と略記することがある):
(1)(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む培地で培養する工程;
[2]培地が、さらに(iii)Wntシグナル阻害剤を含む、上記[1]記載の増殖方法;
[3]膵前駆細胞が、PDX1陽性細胞を(a)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(b)Wntシグナル阻害剤を含む培地で培養することにより誘導された膵前駆細胞である、上記[1]または[2]記載の増殖方法;
[4](i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む、膵前駆細胞の増殖用試薬;
[5](i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む、膵前駆細胞の増殖用キット;
[6]膵前駆細胞を増殖させるための、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤の使用;
[7]膵前駆細胞を、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む培地で培養して増殖させる工程と、培養物中より膵前駆細胞を採取する工程を含む、膵前駆細胞の製造方法;
[8]PDX1陽性細胞を、(a)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(b)Wntシグナル阻害剤を含む培地で培養し、膵前駆細胞に分化誘導させる工程を含む、上記[7]記載の製造方法。
なお、上記[1]〜[8]において、膵前駆細胞およびPDX1陽性細胞は、好ましくはヒト膵前駆細胞およびヒトPDX1陽性細胞である。
以下、本発明および本明細書内で用いられる用語について、説明する。
このうち、「PDX1(pancreatic−duodenal homeobox1)」はinsulin promoter factor 1としても知られ、膵臓の発生およびβ細胞分化に重要な役割を有すると共に生体内の膵β細胞の機能維持にも関与している転写因子である。「NKX6.1」も、PDX1と同様に、β細胞分化に重要な役割を有すると共に生体内の膵β細胞の機能維持にも関与している転写因子である。一方、「INS(INSULIN)」は細胞内インスリンを示し、膵前駆細胞から膵β細胞(インスリン産生細胞)への分化につれて発現が亢進する。
本発明の膵前駆細胞の増殖方法は、膵前駆細胞を、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む培地で培養することを特徴とする。なお、上記増殖方法により膵前駆細胞を増殖させる工程を、本明細書では増殖工程と呼ぶことがある。
培地に含まれるWntシグナル阻害剤の例としては、先に例示した「Wntシグナル阻害剤」が挙げられ、好ましくは2−(4−トリフルオロメチルフェニル)−7,8−ジヒドロ−5H−チオピラノ[4,3−d]ピリミジン−4(3H)−オン(すなわち、XAV939)である。
なお、増殖工程において、培地中のROCK阻害剤をWntシグナル阻害剤に置き換えた場合にも、膵前駆細胞を増殖させることができる。
膵前駆細胞としては、PDX1陽性細胞を(a)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(b)Wntシグナル阻害剤を含む培地で培養することにより誘導された膵前駆細胞を使用することができる。なお、PDX1陽性細胞を上記培地で培養することにより膵前駆細胞を誘導する工程を、本明細書では膵前駆細胞の分化誘導工程と呼ぶことがある。
本発明は、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む膵前駆細胞の増殖用試薬あるいはキットを提供する。
本発明は、膵前駆細胞を増殖させるための、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤の使用も提供する。
上記使用は、(i)と(ii)を組み合わせて、膵前駆細胞を増殖させるために使用するという点に特徴がある。また、上記使用は、(i)と(ii)に、さらにWntシグナル阻害剤を組合せたものであってもよい。使用される膵前駆細胞は特に限定されず、患者由来の細胞あるいは患者由来の細胞から再生させた膵前駆細胞でも、ES細胞やiPS細胞等の多能性幹細胞から誘導した膵前駆細胞であってもよい。
本発明は、膵前駆細胞を、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(ii)ROCK阻害剤を含む培地で培養して増殖させる工程と、培養物中より膵前駆細胞を採取する工程を含む、膵前駆細胞の製造方法を提供する。
本発明の増殖方法あるいは製造方法で得られた膵前駆細胞は、増殖能が高く、機能も保持され、純度も高い。また、フィーダー細胞や他の細胞と共培養をせず、実質的に血清や血清抽出物を含まない培地を使用して培養を行う場合、不純物を含まず、安全性の高い膵前駆細胞が得られる。
ヒトiPS細胞は253G1細胞(レトロウィルスによりOCT4/SOX2/KLF4を発現させて作成されたiPS細胞株;Nature Biotechnology26, 101−106)を使用した。
参考例1にしたがってマトリゲルコートディッシュ上で分化誘導したPDX1陽性細胞を、IMEM−option Zn++培地で洗浄した後、1%のB−27(登録商標)を含むIMEM−option Zn++培地にXAV939(1μM)および/またはbFGF(100ng/ml)(PeproTech)を添加した培地でさらに7日間培養した。
以上の検討により、XAV939とbFGFを添加した培地で培養することにより、効率的に膵前駆細胞を誘導できることが明らかとなった。
参考例1にしたがってマトリゲルコートディッシュ上で分化誘導したPDX1陽性細胞をIMEM−option Zn++培地で洗浄した後、1%のB−27(登録商標)と1μMのXAV939を含むIMEM−option Zn++培地にEGF(500ng/ml)、Betacellulin(40ng/ml)、またはbFGF(100ng/ml)を添加した培地に交換して、8日間培養した。培養後、実施例1に記載の免疫蛍光染色を行い、蛍光顕微鏡で観察した。結果を図2に示す。
297L1細胞(NHDF−iPS、新生児男性の皮膚線維芽細胞にOCT4/SOX2/KLF4/c−MYCを発現させて作製されたヒトiPS細胞株)(PLoS ONE2009; 4(12), p.e8067 参照)を用い、参考例1にしたがってPDX1陽性細胞を誘導した。PDX1陽性細胞をIMEM−option Zn++培地で洗浄した後、1%のB−27(登録商標)を含むIMEM−option Zn++培地にXAV939(1μM)とbFGF(50ng/ml)を添加した培地を用いて、さらに7日間培養した。培養後、実施例1に記載の免疫蛍光染色を行い、蛍光顕微鏡で観察した。結果を図3に示す。
実施例3で誘導した膵前駆細胞を、以下の手順で継代した。すなわち、PBSで洗浄した後、アキュターゼ(Innovative Cell Technologies)を加えて4分間インキュベートし、さらにピペッティングをすることで単一細胞状態とした。IMEM−option Zn++培地で細胞を洗浄した後、継代前の1/4〜1/10の細胞濃度で新しい培養容器に播種した。なお、培地としては、1%のB−27(登録商標)を含むIMEM−option Zn++培地にY27632(10μM)、XAV939(1μM)、およびbFGF(50ng/ml)を添加した培地を用い、培養容器としては、マトリゲルコート処理を施したものを用いた。上記播種後、培地交換を毎日行った。
継代を重ねた増殖膵前駆細胞が正常な核型を保持しているか解析した。28回継代した膵前駆細胞をカルノア固定した後、簡易核型解析(Q−Band)を実施した(株式会社chromocenter)。その結果を図6に示す。その結果、膵前駆細胞は正常な核型を保持しており、長期に培養して継代を重ねても正常な核型が維持されていることが明らかとなった。
実施例4で示したとおり、Y27632、XAV939、bFGFを添加した培地を用いて培養することで膵前駆細胞を安定的に増殖させることが可能であった。そこでこれらの因子のうち、どの因子の組み合わせが膵前駆細胞の増殖に必要であるかを検討した。
他のヒトiPS細胞からも増殖可能な膵前駆細胞を誘導できるか否か検討した。まず、新たなヒトiPS細胞を、以下の方法で作製した。ヘパリンナトリウム含有採血管(テルモ)に採血した血液をPBSで2倍希釈した後、Ficoll−Paque PREMIUM(GE healthcare)に重層して20℃、400gで30分間遠心を行い、peripheral blood mononucleated cell(PBMC)を分離した。Ficollと希釈血液は3:4の比率で使用した。回収したPBMCは、PBSを用いて遠心洗浄を行った後、StemSpan H3000(STEMCELLTechnologies)に再懸濁した。または、セルバンカー3(日本全薬工業)を用いて凍結保存を行った。次に、PBMCを6 well plateに3×106cells/wellの濃度で播種し、10ng/ml IL−3(PeproTech)、100ng/ml IL−6(PeproTech)、300ng/ml SCF(PeproTech)、300ng/ml TPO(PeproTech)、300ng/ml Fit3 ligand(PeproTech)を添加(以下、non−T cell用培地)して6日間培養を行った。培養後増殖したnon−T cellを回収し(約1.3×106 cells/well)、Human CD34 Cell Nucleofector(登録商標) Kit(Lonza)を用いてPlasmids Epi5TM Episomal iPSC Reprogramming Kit(Life Technologies)のエピソーマルベクターを導入した。ベクター量としては、1.3×106 cells当たりReprogramming kit内のEpi5TMReprogramming VectorsおよびEpi5TMp53 & EBNAVectorsをそれぞれ1.5μg(計3μg)使用し、Nucleofector(Lonza)の導入プログラムはU−008を使用した。導入後、細胞をnon−T cell用培地に再懸濁し、Geltrex(Life Technologies)をコートした10−cm dish(培地量10ml)に播種して24時間培養した。翌日(Day1)、1% N2(和光純薬)、2% B−27(登録商標)、1xGlutaMaxI(Life Technologies)、1x NEAA(Life Technologies)および100ng/ml bFGFを含むDMEM/F12(和光純薬)培地を5ml/10−cm dish(計15 ml)加え、その後5日間は同培地で毎日半量交換した。Day9においてEssential 8培地に全量置換し、その後隔日ごとに同培地で培地交換を行った。iPS細胞のコロニーが観察された後、適宜ピックアップし、培養を継続してiPS細胞株を樹立した。
増殖し継代を重ねた膵前駆細胞がINSULIN産生細胞へと分化する能力があるかどうか検討した。細胞としては297L1細胞由来の膵前駆細胞(継代数7)を用いた。アキュターゼを用いて単一細胞状態にした膵前駆細胞を、6x104cells/wellで、マトリゲルコート処理を施した96穴プレートに播種した。培養液は、1%のB−27(登録商標)を含むIMEM−option Zn++培地にXAV939(1μM)、Y27632(10μM)、bFGF(50ng/ml)を添加した培地を用いた。細胞を2日間培養することで細胞密度をコンフルエントにした。その後、細胞をIMEM−option Zn++培地で細胞を洗浄した後、1%のB−27(登録商標)を含むIMEM−option Zn++培地にALK5 inhibitor IIを添加した培地を用いて9日間培養した。ALK5 inhibitor IIはINSULIN陽性細胞を誘導することが知られている(Stem Cell Research 20128, 274−284)。培養後の細胞に対して、4%PFAを添加して室温で30分間の固定を行った。さらに、1次抗体として抗NKX6.1抗体と抗INSULIN抗体(DAKO、A0564)と反応させ、さらに2次抗体としてAlexa488標識2次抗体あるいはAlexa568標識2次抗体と順次反応させた後、蛍光顕微鏡で観察した。その結果を図9に示す。ALK5 inhibitor IIを添加して培養することで、INSULIN陽性細胞が出現する様子が観察された。これらの結果より、増殖させた膵前駆細胞はINSULIN陽性細胞への分化能を持った細胞であることが確認された。
Claims (7)
- 膵前駆細胞を以下の工程(1)に付すことを特徴とする、膵前駆細胞の増殖方法:
(1)(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、(ii)ROCK阻害剤、ならびに(iii)Wntシグナル阻害剤を含む培地で培養する工程であって、Wntシグナル阻害剤がIWP2、IWP3、IWP4、2−(4−トリフルオロメチルフェニル)−7,8−ジヒドロ−5H−チオピラノ[4,3−d]ピリミジン−4(3H)−オン(XAV939)、IWR1、G−CSF、IGFBP4、Dkk1、Cerberus、抗Wnt抗体、Wntアゴニスト(Wnt受容体阻害剤)、及びFrzb−1からなる群から選択される、増殖方法。 - 膵前駆細胞が、PDX1陽性、NKX6.1陰性、かつINSULIN陰性である細胞を(a)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(b)Wntシグナル阻害剤を含む培地で培養することにより誘導された膵前駆細胞である、請求項1記載の増殖方法。
- (i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、(ii)ROCK阻害剤、ならびに(iii)Wntシグナル阻害剤を含む、膵前駆細胞の増殖用試薬であって、Wntシグナル阻害剤がIWP2、IWP3、IWP4、2−(4−トリフルオロメチルフェニル)−7,8−ジヒドロ−5H−チオピラノ[4,3−d]ピリミジン−4(3H)−オン(XAV939)、IWR1、G−CSF、IGFBP4、Dkk1、Cerberus、抗Wnt抗体、Wntアゴニスト(Wnt受容体阻害剤)、及びFrzb−1からなる群から選択される、増殖用試薬。
- (i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、(ii)ROCK阻害剤、ならびに(iii)Wntシグナル阻害剤を含む、膵前駆細胞の増殖用キットであって、Wntシグナル阻害剤がIWP2、IWP3、IWP4、2−(4−トリフルオロメチルフェニル)−7,8−ジヒドロ−5H−チオピラノ[4,3−d]ピリミジン−4(3H)−オン(XAV939)、IWR1、G−CSF、IGFBP4、Dkk1、Cerberus、抗Wnt抗体、Wntアゴニスト(Wnt受容体阻害剤)、及びFrzb−1からなる群から選択される、増殖用キット。
- 膵前駆細胞を増殖させるための、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、(ii)ROCK阻害剤、ならびに(iii)Wntシグナル阻害剤の使用であって、Wntシグナル阻害剤がIWP2、IWP3、IWP4、2−(4−トリフルオロメチルフェニル)−7,8−ジヒドロ−5H−チオピラノ[4,3−d]ピリミジン−4(3H)−オン(XAV939)、IWR1、G−CSF、IGFBP4、Dkk1、Cerberus、抗Wnt抗体、Wntアゴニスト(Wnt受容体阻害剤)、及びFrzb−1からなる群から選択される、使用。
- 膵前駆細胞を、(i)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、(ii)ROCK阻害剤、ならびに(iii)Wntシグナル阻害剤を含む培地で培養して増殖させる工程と、培養物中より膵前駆細胞を採取する工程とを含む、膵前駆細胞の製造方法であって、Wntシグナル阻害剤がIWP2、IWP3、IWP4、2−(4−トリフルオロメチルフェニル)−7,8−ジヒドロ−5H−チオピラノ[4,3−d]ピリミジン−4(3H)オン(XAV939)、IWR1、G−CSF、IGFBP4、Dkk1、Cerberus、抗Wnt抗体、Wntアゴニスト(Wnt受容体阻害剤)、及びFrzb−1からなる群から選択される、製造方法。
- PDX1陽性、NKX6.1陰性、かつINSULIN陰性である細胞を、(a)EGFシグナル伝達活性化因子および/またはFGFシグナル伝達活性化因子、ならびに(b)Wntシグナル阻害剤を含む培地で培養し、膵前駆細胞に分化誘導させる工程を含む、請求項6記載の製造方法。
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Families Citing this family (25)
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KR102523912B1 (ko) | 2013-06-11 | 2023-04-21 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | SC-β 세포 및 조성물 그리고 그 생성 방법 |
US10190096B2 (en) | 2014-12-18 | 2019-01-29 | President And Fellows Of Harvard College | Methods for generating stem cell-derived β cells and uses thereof |
WO2016100930A1 (en) | 2014-12-18 | 2016-06-23 | President And Fellows Of Harvard College | Methods for generating stem cell-derived b cells and methods of use thereof |
US10443042B2 (en) | 2014-12-18 | 2019-10-15 | President And Fellows Of Harvard College | Serum-free in vitro directed differentiation protocol for generating stem cell-derived beta cells and uses thereof |
WO2017188378A1 (ja) * | 2016-04-28 | 2017-11-02 | 武田薬品工業株式会社 | 多能性幹細胞由来膵前駆細胞の純化法とその増幅法 |
US20200140826A1 (en) * | 2017-01-17 | 2020-05-07 | Agency For Science, Research And Technology | Maintenance and Expansion of Pancreatic Progenitor Cells |
BR112020009275A2 (pt) | 2017-11-15 | 2020-10-27 | Semma Therapeutics, Inc. | composições de fabricação de célula de ilhota e métodos de uso |
JP7451430B2 (ja) * | 2018-03-02 | 2024-03-18 | バーテックス ファーマシューティカルズ インコーポレイテッド | 幹細胞のベータ細胞への分化を促進する方法 |
US20210060210A1 (en) | 2018-03-19 | 2021-03-04 | Kyoto University | Hydrogel capsule |
EP3812456A4 (en) | 2018-04-23 | 2022-01-12 | Kyoto University | GROWTH INHIBITOR |
EP3801577A4 (en) * | 2018-05-31 | 2021-07-14 | University Health Network (UHN) | METHOD AND COMPOSITIONS USING TANKYRASE INHIBITOR FOR GENERATING INSULIN-PRODUCING CELLS |
CN112912490B (zh) | 2018-08-03 | 2023-12-26 | 千纸鹤治疗公司 | 细胞制造方法 |
WO2020033879A1 (en) | 2018-08-10 | 2020-02-13 | Semma Therapeutics, Inc. | Stem cell derived islet differentiation |
SG11202102748WA (en) | 2018-09-19 | 2021-04-29 | Takeda Pharmaceuticals Co | Insulin-producing cells |
JPWO2020080550A1 (ja) * | 2018-10-15 | 2021-10-14 | エヴィア ライフ サイエンシズ インコーポレイテッド | 低分子化合物による内胚葉組織又は器官由来細胞からの幹/前駆細胞の作製方法 |
CN109749986B (zh) * | 2019-03-13 | 2021-04-02 | 武汉大学 | 一种由人多能干细胞分化获得胰腺前体细胞及胰岛β细胞的方法 |
CN113993528A (zh) | 2019-04-10 | 2022-01-28 | 千纸鹤治疗公司 | 类生体组织结构体的制造方法 |
JP7385244B2 (ja) * | 2019-06-27 | 2023-11-22 | 国立大学法人 東京大学 | 膵前駆細胞の分離方法 |
CA3158631A1 (en) * | 2019-10-21 | 2021-04-29 | Orizuru Therapeutics, Inc. | Growth inhibitor |
WO2021241668A1 (ja) | 2020-05-28 | 2021-12-02 | 武田薬品工業株式会社 | 均一なサイズの細胞凝集体の大量製造方法 |
JPWO2022107877A1 (ja) | 2020-11-20 | 2022-05-27 | ||
US11970713B2 (en) * | 2020-12-04 | 2024-04-30 | Ocgene Therapeutics Corporation | Method for long-term ex vivo maintenance or expansion of human erythroblast, human megakaryocyte-erythroid progenitor, or human common myeloid progenitor cell and application thereof |
EP4293105A1 (en) | 2021-02-09 | 2023-12-20 | Orizuru Therapeutics, Inc. | Maturation agent |
CN113234664B (zh) * | 2021-05-11 | 2024-05-10 | 澳门大学 | 一种胰腺祖细胞的制备方法及其应用 |
WO2024070494A1 (ja) * | 2022-09-26 | 2024-04-04 | 国立大学法人京都大学 | 膵内胚葉細胞の製造方法 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1175487A2 (en) | 1999-02-10 | 2002-01-30 | Curis, Inc. | Pancreatic progenitor cells, methods and uses related thereto |
US7625753B2 (en) | 2003-12-23 | 2009-12-01 | Cythera, Inc. | Expansion of definitive endoderm cells |
AU2007207434B2 (en) | 2006-01-20 | 2012-04-05 | Reneuron, Inc. | Method of improving cell proliferation of pancreatic progenitor cells in a pancreatic cell culture |
EP2356213B1 (en) * | 2008-11-04 | 2019-05-29 | Viacyte, Inc. | Stem cell aggregate suspension compositions and methods for differentiation thereof |
US8008075B2 (en) * | 2008-11-04 | 2011-08-30 | Viacyte, Inc. | Stem cell aggregate suspension compositions and methods of differentiation thereof |
US9109245B2 (en) * | 2009-04-22 | 2015-08-18 | Viacyte, Inc. | Cell compositions derived from dedifferentiated reprogrammed cells |
EP2505639B1 (en) | 2009-12-29 | 2018-09-19 | Takeda Pharmaceutical Company Limited | Method for manufacturing pancreatic-hormone-producing cells |
EP2909312A4 (en) * | 2012-10-19 | 2016-06-22 | Agency Science Tech & Res | METHOD FOR DIFFERENTIATING STEM CELLS INTO ONE OR MORE CELL LINES |
RU2675928C2 (ru) | 2013-02-06 | 2018-12-25 | Виасайт, Инк. | Композиции клеток, полученные из дедифференцированных перепрограммированных клеток |
JPWO2015125926A1 (ja) * | 2014-02-21 | 2017-03-30 | 国立研究開発法人理化学研究所 | 栄養膜幹細胞の樹立及び維持方法 |
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EP3178924A4 (en) | 2018-01-24 |
EP3178924A1 (en) | 2017-06-14 |
EP3178924B1 (en) | 2019-12-25 |
AU2015300020A1 (en) | 2017-02-23 |
CA2957000C (en) | 2023-08-01 |
CN106795487B (zh) | 2021-11-19 |
JPWO2016021734A1 (ja) | 2017-05-18 |
KR20170031254A (ko) | 2017-03-20 |
CN106795487A (zh) | 2017-05-31 |
KR102416647B1 (ko) | 2022-07-05 |
WO2016021734A1 (ja) | 2016-02-11 |
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