Direct reprogramming of mouse fibroblasts to cardiomyocyte-like cells using Yamanaka factors on engineered poly(ethylene glycol) (PEG) hydrogels
被引:44
作者:
Smith, Amanda W.
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Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Smith, Amanda W.
[1
,2
]
Hoyne, Jake D.
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Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Hoyne, Jake D.
[1
,2
]
Nguyen, Peter K.
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Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Nguyen, Peter K.
[1
,2
]
McCreedy, Dylan A.
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Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
McCreedy, Dylan A.
[1
,2
]
Aly, Haytham
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Washington Univ, Dept Internal Med, Sch Med, St Louis, MO 63124 USA
Washington Univ, Dept Dev Biol, Sch Med, St Louis, MO 63124 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Aly, Haytham
[3
,4
]
Efimov, Igor R.
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Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Efimov, Igor R.
[1
,2
]
Rentschler, Stacey
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Washington Univ, Dept Internal Med, Sch Med, St Louis, MO 63124 USA
Washington Univ, Dept Dev Biol, Sch Med, St Louis, MO 63124 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Rentschler, Stacey
[3
,4
]
Elbert, Donald L.
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Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USAWashington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
Elbert, Donald L.
[1
,2
]
机构:
[1] Washington Univ, Dept Biomed Engn, St Louis, MO 63130 USA
[2] Washington Univ, Ctr Mat Innovat, St Louis, MO 63130 USA
[3] Washington Univ, Dept Internal Med, Sch Med, St Louis, MO 63124 USA
[4] Washington Univ, Dept Dev Biol, Sch Med, St Louis, MO 63124 USA
Direct reprogramming strategies enable rapid conversion of somatic cells to cardiomyocytes or cardiomyocyte-like cells without going through the pluripotent state. A recently described protocol couples Yamanaka factor induction with pluripotency inhibition followed by BMP4 treatment to achieve rapid reprogramming of mouse fibroblasts to beating cardiomyocyte-like cells. The original study was performed using Matrigel-coated tissue culture polystyrene (TCPS), a stiff material that also non-specifically adsorbs serum proteins. Protein adsorption-resistant poly(ethylene glycol) (PEG) materials can be covalently modified to present precise concentrations of adhesion proteins or peptides without the unintended effects of non-specifically adsorbed proteins. Here, we describe an improved protocol that incorporates custom-engineered materials. We first reproduced the Efe et al. protocol on Matrigel-coated TCPS (the original material), reprogramming adult mouse tail-tip mouse fibroblasts (TTF) and mouse embryonic fibroblasts (MEF) to cardiomyocyte-like cells that demonstrated striated sarcomeric alpha-actinin staining, spontaneous calcium transients, and visible beating. We then designed poly(ethylene glycol) culture substrates to promote MEF adhesion via laminin and RGD-binding integrins. PEG hydrogels improved proliferation and reprogramming efficiency (evidenced by beating patch number and area, gene expression, and flow cytometry), yielding almost twice the number of sarcomeric alpha-actinin positive cardiomyocyte-like cells as the originally described substrate. These results illustrate that cellular reprogramming may be enhanced using custom-engineered materials. (C) 2013 Elsevier Ltd. All rights reserved.
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Ieda, Masaki
Fu, Ji-Dong
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Fu, Ji-Dong
Delgado-Olguin, Paul
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Delgado-Olguin, Paul
Vedantham, Vasanth
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Med, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Vedantham, Vasanth
Hayashi, Yohei
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Hayashi, Yohei
Bruneau, Benoit G.
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Bruneau, Benoit G.
Srivastava, Deepak
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Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Jayawardena, Tilanthi M.
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Egemnazarov, Bakytbek
Finch, Elizabeth A.
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Ion Channel Res Grp, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Finch, Elizabeth A.
Zhang, Lunan
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Zhang, Lunan
Payne, J. Alan
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Payne, J. Alan
Pandya, Kumar
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Univ N Carolina, Dept Pathol & Lab Med, Chapel Hill, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Pandya, Kumar
Zhang, Zhiping
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Zhang, Zhiping
Rosenberg, Paul
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Ion Channel Res Grp, Durham, NC USA
Duke Univ, Sch Med, Sarah Steadman Nutr & Metab Ctr, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Rosenberg, Paul
Mirotsou, Maria
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Mirotsou, Maria
Dzau, Victor J.
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Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Ieda, Masaki
Fu, Ji-Dong
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Fu, Ji-Dong
Delgado-Olguin, Paul
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h-index: 0
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Delgado-Olguin, Paul
Vedantham, Vasanth
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h-index: 0
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Med, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Vedantham, Vasanth
Hayashi, Yohei
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h-index: 0
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Hayashi, Yohei
Bruneau, Benoit G.
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h-index: 0
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Bruneau, Benoit G.
Srivastava, Deepak
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h-index: 0
机构:
Univ Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USAUniv Calif San Francisco, Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Jayawardena, Tilanthi M.
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机构:
Egemnazarov, Bakytbek
Finch, Elizabeth A.
论文数: 0引用数: 0
h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Ion Channel Res Grp, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Finch, Elizabeth A.
Zhang, Lunan
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h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Zhang, Lunan
Payne, J. Alan
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h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Payne, J. Alan
Pandya, Kumar
论文数: 0引用数: 0
h-index: 0
机构:
Univ N Carolina, Dept Pathol & Lab Med, Chapel Hill, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Pandya, Kumar
Zhang, Zhiping
论文数: 0引用数: 0
h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Zhang, Zhiping
Rosenberg, Paul
论文数: 0引用数: 0
h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Ion Channel Res Grp, Durham, NC USA
Duke Univ, Sch Med, Sarah Steadman Nutr & Metab Ctr, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Rosenberg, Paul
Mirotsou, Maria
论文数: 0引用数: 0
h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Mirotsou, Maria
Dzau, Victor J.
论文数: 0引用数: 0
h-index: 0
机构:
Duke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA
Duke Univ, Med Ctr, Mandel Ctr Hypertens & Atherosclerosis Res, Durham, NC USADuke Univ, Med Ctr, Duke Cardiovasc Res Ctr, Durham, NC USA