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Scalable Producing Embryoid Bodies by Rotary Cell Culture System and Constructing Engineered Cardiac Tissue with ES‐Derived Cardiomyocytes in Vitro
Author(s) -
Wang Xiuli,
Wei Guofeng,
Yu Weiting,
Zhao Yunshan,
Yu Xingju,
Ma Xiaojun
Publication year - 2006
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp060018z
Subject(s) - petri dish , percoll , microbiology and biotechnology , embryoid body , embryonic stem cell , cell culture , cellular differentiation , cell , in vitro , cell type , mesenchymal stem cell , biology , confocal microscopy , chemistry , biomedical engineering , biochemistry , adult stem cell , genetics , gene , medicine
Abstract Embryonic stem (ES) cells are of significant interest either as an in vitro model recapitulating early embryonic development or as a renewable source of therapeutically useful cells. ES cells aggregation is important for embryoid bodies (EBs) formation and the subsequent generation of ES cell derivatives. This study was conducted to describe scalable production of EBs by the rotary cell culture system (RCCS, STLV type) and estimate the feasibility of constructing engineered cardiac tissue (ECT). In comparison with suspension culture in a Petri dish, the efficiency of the dynamic process was analyzed with respect to the yield of EB formation and their cardiomyocyte differentiation. Cardiomyocyte differentiation was evaluated by immunohistochemical analysis. After the elementary enrichment by gradient percoll, ES cell‐derived cardiomyocytes were applied to construct ECT. Cell gross morphology, spatial distribution, and ultrastructure were evaluated by using histological analysis, confocal laser scanning microscopy, and transmission electron microscopy. Results showed that EB efficiencies in STLV were nearly 1.5–2.0 times higher than that of liquid suspension cultures, and cardiomyocyte differentiation of EBs progressed in a normal course after the dynamic cultivation in STLV. Additionally, the differentiated cultures could be enriched elementarily by gradient percoll. Once cast into the collagen strand, cells grew well and became more matured in Petri dishes. Synchronous contraction of the cell cluster was observed on the surface of the ECT, and cell connection was also established. It was the first report to have beating ES‐derived cardiomyocytes on a 3‐D collagen scaffold, which might provide a promising model for physiological and pharmacological studies and tissue replacement therapy.