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. [1 ,2 ]
Hoyne, Jake D. [1 ,2 ]
Nguyen, Peter K. [1 ,2 ]
McCreedy, Dylan A. [1 ,2 ]
Aly, Haytham [3 ,4 ]
Efimov, Igor R. [1 ,2 ]
Rentschler, Stacey [3 ,4 ]
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
基金
美国国家卫生研究院;
关键词
Direct reprogramming; Stem cell niche; Cardiomyocyte; poly(ethylene glycol); Laminin; Stem cell microenvironment; PLURIPOTENT STEM-CELLS; ACUTE MYOCARDIAL-INFARCTION; SELF-RENEWAL; IN-VITRO; FUNCTIONAL CARDIOMYOCYTES; SPHINGOSINE; 1-PHOSPHATE; CARDIAC DIFFERENTIATION; BIOLOGICAL-ACTIVITY; DEFINED CONDITIONS; INTEGRIN BINDING;
D O I
10.1016/j.biomaterials.2013.05.050
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
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.
引用
收藏
页码:6559 / 6571
页数:13
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