期刊
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
卷 86A, 期 3, 页码 713-724出版社
WILEY
DOI: 10.1002/jbm.a.31578
关键词
cardiac tissue engineering; myocardium; fibroblast; cardiomyocyte; collagen; scaffold
资金
- NHLBI NIH HHS [R01 HL076485, R01 HL076482, R01 HL076485-04] Funding Source: Medline
- NIBIB NIH HHS [P41 EB002520, P41 EB002520-049002] Funding Source: Medline
Native myocardium consists of several cell types, of which approximately one-third are myocytes and most of the nonmyocytes are fibroblasts. By analogy with monolayer culture in which fibroblasts were removed to prevent over-growth, early attempts to engineer myocardium utilized cell populations enriched for cardiac myocytes (CMs; similar to 80-90% of total cells). We hypothesized that the pre-treatment of synthetic elastomeric scaffolds with cardiac fibroblasts (CFs) will enhance the functional assembly of the engineered cardiac constructs by creating an environment supportive of cardiomyocyte attachment and function. Cells isolated from neonatal rat ventricles were prepared to form three distinct populations: rapidly plating cells identified as CFs, slowly plating cells identified as CMs, and unseparated initial population of cells (US). The cell fractions (3 X 106 cells total) were seeded into poly(glycerol sebacate) scaffolds (highly porous discs, 5 mm in diameter X 2-mm thick) using Matrigel (TM), either separately (CM or CF), concurrently (US), or sequentially (CF pre-treatment followed by CM culture, CF + CM), and cultured in spinner flasks. The CF + CM group had the highest amplitude of contraction and the lowest excitation threshold, superior DNA content, and higher glucose consumption rate. The CF + CM group exhibited compact 100- to 200-mu m thick layers of elongated myocytes aligned in parallel over layers of collagen-producing fibroblasts, while US and CM groups exhibited scattered and poorly elongated myocytes. The sequential co-culture of CF and CM on a synthetic elastomer scaffold thus created an environment supportive of cardiomyocyte attachment, differentiation, and contractile function, presumably due to scaffold conditioning by cultured fibroblasts. When implanted over the infarcted myocardium in a nude rat model, cell-free poly(glycerol sebacate) remained at the ventricular wall after 2 weeks of in vivo, and was vascularized. (c) 2007 Wiley Periodicals, Inc.
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