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Efficient cardiomyocyte differentiation of embryonic stem cells by bone morphogenetic protein‐2 combined with visceral endoderm‐like cells
Author(s) -
Bin Zeng,
Sheng LinGuo,
Gang ZhuChen,
Hong Jiang,
Jun Cai,
Bo Yang,
Hui Shen
Publication year - 2006
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2006.05.011
Subject(s) - endoderm , embryonic stem cell , microbiology and biotechnology , bone morphogenetic protein , stem cell , biology , chemistry , anatomy , biochemistry , gene
As the signals required for cardiomyocyte differentiation and functional regulation are complex and only partly understood, the mechanisms prompting the differentiation and specification of pluripotential embryonic stem (ES) cells into cardiomyocytes remain unclear. We hypothesized that a combined technology system, cocultured with a visceral endoderm (VE)—like cell line, END‐2, and added cytokine BMP‐2, would induce high percentage conversion of murine ES‐D3 cell line into cardiomyocytes, and derived cardiomyocytes in this system would exhibit more mature characteristics. It was observed that 92% ( P < 0.01) ES cell‐derived aggregates in this system exhibited rhythmic contractions, and the contractile areas were greater. By contrast, in ES cells cultured alone, on the feeder layer of END‐2 cells, or with added BMP‐2, the total percentage of beating aggregates was 19, 69 ( P < 0.01) and 44% ( P < 0.01), respectively. All the rhythmically contractile cells derived from ES cells expressed cardiac‐specific proteins for troponin T. Among them, the combined system resulted in significantly increased cardiac‐specific genes (NKx2.5, α‐MHC). Transmission electron microscopy (TEM) revealed varying degrees of myofibrillar organization, and the combined system resulted in a more mature phenotype such as Z bands, nascent intercalated discs and gap junctions. Before shifting to the cardiomyocyte phenotype, this system could accelerate apoptosis of the cell population ( P < 0.01). The inductive efficacy of this system can provide an opportunity to facilitate cardiomyocyte differentiation of ES cells. The inducible effects of this system may depend on increasing cardiac‐specific gene expression and the induction of apoptosis in cells that are not committed to cardiac differentiation.